WILL MAKEUN USKULUNUTMIMANTAN 20170296623A1 ( 19) United States (12 ) Patent Application Publication ( 10) Pub . No. : US 2017/ 0296623 A1 JUILLERAT et al. (43 ) Pub . Date : Oct. 19 , 2017 (54 ) INHIBITORY CHIMERIC ANTIGEN Publication Classification RECEPTOR ( ICAR OR N -CAR ) EXPRESSING NON - T CELL TRANSDUCTION DOMAIN (51 ) Int . CI. A61K 38 / 17 ( 2006 .01 ) (71 ) Applicant: CELLECTIS , Paris (FR ) CO7K 14 /47 ( 2006 . 01) COZK 14 / 705 (2006 . 01) (72 ) Inventors : Alexandre JUILLERAT , New York , COZK 14 /525 ( 2006 .01 ) NY (US ); Philippe DUCHATEAU , A61K 38 /00 (2006 . 01 ) Draveil (FR ); Laurent POIROT, Paris CO7K 14 / 00 (2006 . 01) (FR ) 2 ) U . S . CI. CPC ...... A61K 38 / 17 (2013 .01 ) ; CO7K 14 / 70596 ( 21 ) Appl. No. : 15 /532 , 430 (2013 .01 ) ; C07K 14 /525 ( 2013 .01 ) ; CO7K 14 /4748 (2013 . 01 ); CO7K 14 /00 (2013 . 01 ) ; ( 22 ) PCT Filed : Dec. 17, 2015 A61K 38 / 00 ( 2013 .01 ) ( 86 ) PCT No .: PCT/ EP2015 / 080376 $ 371 (c )( 1) , (57 ) ABSTRACT ( 2 ) Date : Jun . 1 , 2017 (30 ) Foreign Application Priority Data The invention relates to negative T - cell signal inducing chimeric antigen receptor ( N -CAR or i -CAR ) and to T -cells Dec . 17 , 2014 (DK ) ...... PA201470797 comprising such N -CAR as well as a positive T - cell signal Aug . 11, 2015 (DK ) ...... PA201570518 inducing CAR ( P -CAR ) as well as their use in therapy.
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: . seorang dalah * *: : . A . . LAERSKABER S1189-1+8V . . 2.: ." "27: "-: -" : :- . . . *. : . : T/I*£1101 : :-%* % 429: *: :. $|25-1 . 7 . WWWXXNDSWORDS . . -27.ver : : *-> :* . *" -.7% °. ! PREHRANA I i o low CAR- N antigen/ high CAR - P antigen and high CAR- N antigen / high CAR - P antigen cells target of % of Ratio US 2017 /0296623 A1 Oct . 19 , 2017 INHIBITORY CHIMERIC ANTIGEN [0007 ] Novel specificities in T cells have been success RECEPTOR (ICAR OR N -CAR ) EXPRESSING fully generated through the genetic transfer of transgenic T NON - T CELL TRANSDUCTION DOMAIN cell receptors or chimeric antigen receptors (CARS ) (Jena , Dotti et al. 2010 , Blood 116 ( 7 ) : 1035 -44 ) . CARs are syn FIELD OF THE INVENTION thetic receptors consisting of a targeting moiety that is associated with one or more signaling domains in a single [0001 ] The invention relates to negative T - cell signal fusion molecule . In general, the binding moiety of a CAR inducing chimeric antigen receptor ( N -CAR or i -CAR ) and consists of an antigen -binding domain of a single - chain to T - cells comprising such N -CAR as well as a positive antibody ( scFv ) , comprising the light and heavy variable T -cell signal inducing CAR ( P -CAR ) as well as their use in fragments of a monoclonal antibody joined by a flexible therapy . In particular, the present invention relates to an linker. Binding moieties based on receptor or ligand immune cell engineered to express at least one N -CAR and domains have also been used successfully . The signaling at least one P -CAR , wherein the P -CAR binds to a first domains for first generation CARs are derived from the antigen and activates the immunoresponsive cell ( i. e . cyto cytoplasmic region of the SCD3zeta or the Fc receptor toxicity ) whereas the N -CAR binds to a second antigen and gamma chains . First generation CARs have been shown to inhibits the immunoresponsive cell ( i. e . cytotoxicity ) successfully redirect T cell cytotoxicity , however, they failed through the signaling of a sequence from a TRAIL receptor to provide prolonged expansion and anti- tumor activity in or a CD200R1 receptor. vivo . Signaling domains from co - stimulatory molecules [0002 ] The invention also relates to negative T -cell signal including CD28 , OX - 40 (CD134 ) , ICOS and 4 - 1BB inducing chimeric antigen receptor ( N -CAR or i -CAR ) and (CD137 ) have been added alone (second generation ) or in to T - cells comprising such N -CAR as well as a positive combination (third generation ) to enhance survival and T - cell signal inducing CAR ( P -CAR ) as well as their use in increase proliferation of CAR modified T cells . CARs have therapy . In particular, the present invention relates to an successfully allowed T cells to be redirected against antigens immune cell engineered to express at least one N -CAR and expressed at the surface of tumor cells from various malig at least one P -CAR , wherein the P -CAR binds to a first nancies including lymphomas and solid tumors ( Jena , Dotti antigen and activates the immunoresponsive cell ( i . e . cyto et al . 2010 , Blood 116 ( 7 ) : 1035 - 44 ) . toxicity ) whereas the N -CAR binds to a second antigen and [0008 ] However, despite their unprecedent efficacy for inhibits the immunoresponsive cell ( i . e . cytotoxicity ) tumor eradication in vivo , CAR T cells can promote acute through the signaling of a sequence usually expressed in non adverse events after being transferred into patients . Among T cells , provided that said sequence is not an ITSM prefer the well documented adverse events is Graft versus host ably not a sequence selected in a group consisting of SEQ ID disease (GVHD ) , on -target off- tumor activity or aberrant NO : 13 (human SIGLS ) , SEQ ID NO : 14 (human SIGL7) , lymphoproliferative capacity due to vector derived inser SEQ ID NO : 17 ( human SIGL5 ) , SEQ ID NO : 20 (human tionalmutagenesis . Therefore , there is a need to develop cell SIGL9 ), SEQ ID NO : 21 ( human SIGLO ), SEQ ID NO : 22 specific depletion systems to prevent such deleterious events (human CD33 ), SEQ ID NO : 26 (human SIG12 ) , SEQ ID to occur in vivo . NO :31 (human SIG11) , SEQ ID NO :32 (human SIG10 ) and [0009 ] Recently , inhibitory chimeric antigen receptors SEQ ID NO : 19 (human PECA1) . ( N - CARs) were designed having as objective to put the [ 0003 ] or brakes on T cell function upon encountering off - target cells [ 0004 ] provided that said sequence does not comprise a ( Fedorov , V . D . , Themeli, M . , Sadelain , M , 2013 , Sci Transl sequence selected in a group consisting of SEQ ID NO : 13 Med 5 (215 ) . In this paper, the authors designed CLTA - 4 (human SIGL8 ), SEO ID NO : 14 (human SIGL7 ) , SEO ID and PD - 1 - based N -CARs which could selectively limit NO : 17 (human SIGL5 ) , SEQ ID NO :20 (human SIGL9) , cytokine secretion , cytotoxicity , and proliferation induced SEQ ID NO : 21 (human SIGL6 ) , SEQ ID NO :22 (human through the endogenous T cell receptor or an activating CD33 ) , SEQ ID NO : 26 (human SIG12 ) , SEQ ID NO :31 chimeric receptor. They have shown that the initial effect of (human SIG11 ) , SEQ ID NO :32 (human SIG10 ) and SEQ the N - CAR is temporary , thus enabling T cells to function ID NO : 19 (human PECA1) . upon a subsequent encounter with the antigen recognized by [ 0005 ] This system of “ NOT gates” is particularly useful their activating receptor . in immunotherapy in order to prevent cytotoxicity towards [0010 ] Proteins containing Immunoreceptor tyrosine " off - target” healthy or immune cells. based inhibitory motif (ITIM ) , immunoreceptor tyrosine based switch motif (ITSM ) and SH2- binding motif are, as non - limiting example , known to play a major role in the INTRODUCTION inhibition , control and modulation of several signaling path [ 0006 ] Adoptive immunotherapy, which involves the ways in T -cells ( e . g . TCR ) (Barrow Aand Trowsdale J , 2006 , transfer of autologous antigen - specific T cells generated ex Eur J Immunol 36 (7 ) : 1646 - 53 , Sharpe H and Freeman G , vivo , is a promising strategy to treat viral infections and 2002, Nature Reviews Immunology , (2 ) 116 - 126 ) . cancer . The T cells used for adoptive immunotherapy can be [0011 ] However, when the engineered iCAR or N -CAR generated either by expansion of antigen - specific T cells or T - cells contain a transduction domain from T - cell , it may redirection of T cells through genetic engineering (Park , occur an interference or protein interaction with the trans Rosenberg et al. 2011 , Trends Biotechnol 29 ( 11) :550 - 7 ) . duction domain from wild - type T - cells . In order to circum Transfer of viral antigen specific T cells is a well -established vent this problem , the inventors have sought engineered procedure used for the treatment of transplant associated inhibitory chimeric antigen receptor ( N -CAR ) based on viral infections and rare viral- related malignancies . Simi non -naturally expressed intracellular domains in T- cell and / larly , isolation and transfer of tumor specific T cells has been or intracellular domain from TRAIL receptors and/ or CD200 shown to be successful in treating melanoma. receptor 1 . This N -CAR can be used in “ logic NOT gates " US 2017 /0296623 A1 Oct . 19 , 2017 systems which are composed of a positive signaling CAR polynucleotides and vectors encoding said CARs, as well as ( P -CAR ) and an inhibitory Gate receptor ( iCAR or N -CAR ) . therapeutic treatment using such engineered immune cell . SUMMARY BRIEF DESCRIPTION OF DRAWINGS [0015 ] FIG . 1 : Schematic representation of the architec [ 0012 ] The present invention is drawn to apply biology tures ( versions V1 to V6 , preferably V1, V3 and V5 ) for the principles such as logic “ NOT gate ” to immune cell tech different single -chain car chimeric antigen receptor (scCAR ) nology in order for the engineered immune cells , in particu of the invention . lar T - cells , to be inhibited in case off - tumor targets ( healthy [00161 FIG . 2 : Schematic representation of the design of cells ). In particular, the present invention relates to an inhibitory Gate receptors ( N -CAR ): the native signaling inhibitory chimeric antigen receptor ( iCAR or N - CAR ) modulation receptor is engineered in order to replace the which contains an intracellular domain from a receptor native extracellular topological domain by an extracellular involved in transduction signal which is not significantly binding domain able to bind specifically to an antigen or cell expressed in natural T - cell and / or from a TRAIL receptor surface marker of an “ off - target ” healthy cell . and / or the CD200 receptor . The preferred intracellular [0017 ] FIG . 3 : P -CAR ( P is CD20 ) driven activation domains of the invention have at least 80 % identity with the (measured by expression of CD69 ) of transduced T cells polypeptides of SEQ ID NO : 1 to 36 . More preferably an mediated through target cells expressing the CD20 antigen . intracellular domain of the CAR P of the invention com CD69 is an appropriate marker for measuring T cell activa prises a sequence selected from the group consisting ofSEQ tion . ID NO : 1 to 36 . [0018 ] FIG . 4 : FIG . 4 shows Ratio of % of target cells [0013 ] Another aspect of the invention is the engineered antigen P -CAR -high /antigen N -CAR -high and antigen immune cell such as T -cell which expressed both said P -CAR -high / antigen N -CAR - low after a co - incubation of 6 N -CAR and a positive CAR ( P -CAR ) ; their respective h with engineered primary T -cells ( three ratio of target / extracellular binding domains targeting an off tumor cell effectors are used : 1 / 1 , 1 / 3 and 1 /10 ) . (healthy cell ) and a tumoral cell. [0014 ] The present invention also relates to a method of DETAILED DESCRIPTION engineering of such N - CAR and isolated immune cell, [0019 ] Abreviations Designation N -CAR Or iCAR negative chimeric antigen receptor P -CAR positive chimeric antigen receptor TRAIL tumor necrosis - factor related apoptosis inducing ligand CDR complementarity determining regions scFv single chain antibody fragment KIR2DL2 CD158B1 NKAT6 Killer cell immunoglobulin - like receptor 2DL2 (CD158 antigen - like family member B1) (MHC class I NK cell receptor ) (Natural killer associated transcript 6 ) (NKAT - 6 ) (p58 natural killer cell receptor clone CL - 43 ) (p58 NK receptor CL -43 ) (CD antigen CD15861) KIR2DL1 CD158A NKAT1 Killer cell immunoglobulin - like receptor 2DL1 (CD158 antigen -like family member A ) (MHC class I NK cell receptor) (Natural killer associated transcript 1 ) (NKAT - 1 ) (p58 natural killer cell receptor clones CL -42 /47 . 11) (p58 NK receptor CL -42 /47 . 11) (p58 . 1 MHC class - I specific NK receptor ) ( CD antigen CD158a ) FCGR2B CD32 FCG2 IGFR2 Low affinity immunoglobulin gamma Fc region receptor II- b ( IgG Fc receptor II - b ) (CDw32 ) ( Fc -gamma RII - b ) (Fc - gamma- Rib ) ( FcRII- b ) (CD antigen CD32 ) KIR2DL3 CD158B2 KIRCL23 Killer cell immunoglobulin - like receptor 2DL3 ( CD158 antigen - like NKAT2 family member B2 ) (KIR -023GB ) (Killer inhibitory receptor cl 2 - 3 ) (MHC class I NK cell receptor) (NKAT2a ) (NKAT2b ) (Natural killer- associated transcript 2 ) (NKAT - 2 ) (p58 natural killer cell receptor clone CL - 6 ) (p58 NK receptor CL - 6 ) (p58 . 2 MHC class- I - specific NK receptor) (CD antigen CD15862) KIR3DL2 CD158K NKAT4 Killer cell immunoglobulin - like receptor 3DL2 (CD158 antigen - like family member K ) MHC( class I NK cell receptor) ( Natural killer associated transcript 4 ) (NKAT - 4 ) (p70 natural killer cell receptor clone CL - 5 ) (p70 NK receptor CL - 5 ) (CD antigen CD158k ) KIR2DL4 CD158D KIR103AS Killer cell immunoglobulin - like receptor 2DL4 (CD158 antigen - like family member D ) (G9P ) (Killer cell inhibitory receptor 103AS ) (KIR 103AS ) (MHC class I NK cell receptor KIR103AS ) ( CD antigen CD158d ) KIR3 DL1 CD158E NKAT3 Killer cell immunoglobulin - like receptor 3DL1 (CD158 antigen - like NKB1 family member E ) (HLA - BW 4 -specific inhibitory NK cell receptor ) (MHC class I NK cell receptor ) ( Natural killer- associated transcript 3 ) (NKAT 3 ) (p70 natural killer cell receptor clones CL - 2 / CL - 11 ) (p70 NK receptor CL- 2/ CL - 11) ( CD antigen CD158e) KIR2DL5A CD158F Killer cell immunoglobulin - like receptor 2DL5A (CD antigen CD158f1 ) CD158F1 KIR2DL5 MILR1 C17orf60 MCA32 Allergin - 1 (Allergy inhibitory receptor 1 ) (Mast cell antigen 32 ) (MCA 32 ) (Mast cell immunoglobulin - like receptor 1 ) US 2017 /0296623 A1 Oct . 19 , 2017 - continued Abreviations Designation LILRB4 ILT3 LIR5 Leukocyte immunoglobulin - like receptor subfamily B member 4 (CD85 antigen - like family member K ) ( Immunoglobulin - like transcript 3 ) ( ILT 3) ( Leukocyte immunoglobulin - like receptor 5 ) (LIR - 5 ) (Monocyte inhibitory receptor HM18 ) ( CD antigen CD85k ) LILRB3 ILT5 LIR3 Leukocyte immunoglobulin - like receptor subfamily B member 3 (LIR - 3 ) (Leukocyte immunoglobulin - like receptor 3 ) (CD85 antigen - like family member A ) ( Immunoglobulin - like transcript 5 ) ( ILT- 5 ) (Monocyte inhibitory receptor HL9 ) ( CD antigen CD85a ) KIR3DL3 CD158Z KIR3DL7 Killer cell immunoglobulin - like receptor 3DL3 (CD158 antigen - like KIRC1 family member Z ) (Killer cell inhibitory receptor 1) ( CD antigen CD1582 ) SIGLEC8 SAF2 Sialic acid -binding Ig - like lectin 8 (Siglec -8 ) (Sialoadhesin family member 2 ) ( SAF - 2 ) SIGLEC7 AIRM1 Sialic acid - binding Ig -like lectin 7 (Siglec - 7 ) (Adhesion inhibitory receptor molecule 1 ) (AIRM - 1 ) ( CDw328 ) ( D - siglec ) (QA79 membrane protein ) (p75 ) (CD antigen CD328 ) LILRB5 LIRS Leukocyte immunoglobulin - like receptor subfamily B member 5 (CD85 antigen - like family member C ) (Leukocyte immunoglobulin - like receptor 8 ) ( LIR - 8 ) ( CD antigen CD85c ) LILRB2 ILT4 LIR2 MIR10 Leukocyte immunoglobulin - like receptor subfamily B member 2 (LIR - 2 ) (Leukocyte immunoglobulin - like receptor 2 ) (CD85 antigen - like family member D ) ( Immunoglobulin - like transcript 4 ) ( ILT- 4 ) (Monocyte /macrophage immunoglobulin - like receptor 10 ) (MIR - 10 ) (CD antigen CD85d ) SIGLEC5 CD33L2 OBBP2 Sialic acid -binding Ig - like lectin 5 (Siglec - 5 ) ( CD33 antigen - like 2 ) (Obesity -binding protein 2 ) (OB - BP2 ) ( OB - binding protein 2 ) (CD antigen CD170 ) FCRL4 FCRH4 IFGP2 IRTA1 Fc receptor- like protein 4 ( FcR - like protein 4 ) (FCRL4 ) (Fc receptor homolog 4 ) (FcRH4 ) ( IFGP family protein 2 ) (hIFGP2 ) ( Immune receptor translocation -associated protein 1 ) (CD antigen CD307d ) PECAM1 Platelet endothelial cell adhesion molecule (PECAM - 1 ) (EndoCAM ) (GPIIA ') ( PECA1) (CD antigen CD31) SIGLEC9 Sialic acid -binding Ig -like lectin 9 ( Siglec- 9 ) (CDw329 ) (Protein FOAP- 9 ) UNQ668/ PRO1302 (CD antigen CD329 ) SIGLEC6 CD33L CD33L1 Sialic acid - binding Ig - like lectin 6 ( Siglec - 6 ) (CD33 antigen - like 1 ) OBBP1 ( CDw327 ) ( Obesity -binding protein 1 ) (OB - BP1) (CD antigen CD327 ) CD33 SIGLEC3 Myeloid cell surface antigen CD33 (Sialic acid - binding Ig - like lectin 3 ) (Siglec - 3 ) ( gp67 ) (CD antigen CD33 ) FCRL5 FCRH5 IRTA2 Fc receptor - like protein 5 (FcR - like protein 5 ) (FcRL5 ) (BXMASI ) (Fc UNQ503 /PRO820 receptor homolog 5 ) (FcRH5 ) ( Immune receptor translocation associated protein 2 ) (CD antigen CD307e ) FCRL2 FCRH2 IFGP4 IRTA4 Fc receptor - like protein 2 ( FcR - like protein 2 ) (FcRL2 ) ( Fc receptor SPAP1 homolog 2 ) (FcRH2 ) (IFGP family protein 4 ) ( Immunoglobulin receptor UNQ9236 / PRO31998 translocation -associated protein 4 ) ( SH2 domain -containing phosphatase anchor protein 1 ) (CD antigen CD307b ) FCRL1 FCRH1 IFGP1 IRTA5 Fc receptor- like protein 1 (FcR - like protein 1 ) (FcRL1 ) (Fc receptor homolog 1 ) (FcRH1 ) ( IFGP family protein 1 ) (hIFGP1 ) ( Immune receptor translocation - associated protein 5 ) (CD antigen CD307a ) SIGLEC12 SIGLECL1 SLG Sialic acid - binding Ig - like lectin 12 (Siglec - 12 ) (Sialic acid -binding Ig - like UNQ9215 / PRO34042 lectin - like 1 ) ( Siglec -L1 ) FCRL3 FCRH3 IFGP3 IRTA3 Fc receptor - like protein 3 ( FcR - like protein 3 ) (FcRL3 ) ( Fc receptor SPAP2 homolog 3 ) ( FcRH3 ) ( IFGP family protein 3 ) ( hIFGP3 ) ( Immune receptor translocation - associated protein 3 ) (SH2 domain -containing phosphatase anchor protein 2 ) (CD antigen CD307c ) MPZL1 PZR Myelin protein zero - like protein 1 (Protein zero -related ) UNQ849 /PRO1787 PILRA Paired immunoglobulin -like type 2 receptor alpha (Cell surface receptor FDF03) ( Inhibitory receptor PILR - alpha ) PVR PVS Poliovirus receptor (Nectin - like protein 5 ) (NECL -5 ) ( CD antigen CD155 ) SIGLEC11 Sialic acid - binding Ig - like lectin 11 (Sialic acid -binding lectin 11 ) ( Siglec UNQ9222 /PRO28718 11 ) SIGLEC10 SLG2 Sialic acid - binding Ig -like lectin 10 (Siglec - 10 ) (Siglec - like protein 2 ) UNQ477 / PRO940 TNFRSF10D DCR2 TRAILR4 Tumor necrosis factor receptor superfamily member 10D (Decoy TRUNDD UNQ251/ PRO288 receptor 2 ) (DcR2 ) ( TNF - related apoptosis - inducing ligand receptor 4 ) ( TRAIL receptor 4 ) ( TRAIL - R4 ) ( TRAIL receptor with a truncated death domain ) (CD antigen CD264 ) TNFRSF10A APO2 DR4 Tumor necrosis factor receptor superfamily member 10A ( Death TRAILR1 receptor 4 ) ( TNF - related apoptosis - inducing ligand receptor 1 ) ( TRAIL receptor 1 ) ( TRAIL - R1 ) ( CD antigen CD261) TNFRSF10B DR5 KILLER Tumor necrosis factor receptor superfamily member 10B (Death TRAILR2 TRICK2 ZTNFR9 receptor 5 ) ( TNF -related apoptosis - inducing ligand receptor 2 ) ( TRAIL UNQ160 / PRO186 receptor 2 ) ( TRAIL -R2 ) ( CD antigen CD262 ) US 2017 /0296623 A1 Oct . 19 , 2017 -continued |Abreviations Designation CD200R1 CD200R CRTR2 Cell surface glycoprotein CD200 receptor 1 (CD200 cell surface MOX2R OX2R glycoprotein receptor ) (Cell surface glycoprotein OX2 receptor 1 ) UNQ2522 /PRO6015 CD " XX " cluster of differentiation RARRES1 Retinoic Acid Receptor Responder ( Tazarotene Induced ) 1 ) CCKBR Cholecystokinin B Receptor GALR1 galanin receptor 1 CUBN Cubilin MUC1 Mucin 1 5T4 Trophoblast glycoprotein , also known as TPBG ROR1 orphan -receptor tyrosine- kinase - like surface Nkp30 Natural cytotoxicity receptors (Synonym CD337 ) NKG2D Killer cell lectin - like receptor subfamily K , member 1 CS1 SLAM family member 7 MART1 Antigen LB39- AA , called also Antigen SK29 - AA WT1 Wilms tumor protein LMP2 latent membrane protein 2 gp100 Glycoprotein 100 or Melanocyte protein PMEL bcr -abl called also BCR /ABL fusion protein isoform X8 hTERT Telomerase transcriptase EphA2 Eph receptor A2 ERG PAX3 Paired box protein Pax - 3 PD - 1 Programmed cell death protein 14 - 1BB OX40 tumor necrosis factor ligand superfamily member 4 PSMA Prostate specific membrane antigen ICOS Inducible T - cell costimulator CTLA - 4 Cytotoxic T - lymphocyte protein 4 LAGU Lymphocyte activation gene 3 protein 2B4 : Natural killer cell receptor 2B4 (CD244 CTLA - 4 Cytotoxic T- lymphocyte protein 4 TIM - 3 Protein timeless TIGIT Protein Tigit SIRPA Tyrosine -protein phosphatase non - receptor type substrate 1 ALK ALK tyrosine kinase Endoglin also called CD105 PD - L1 Programmed cell death 1 ligand 1 PD - L2 Programmed cell death 1 ligand 2 ICAM Intercellular adhesion molecule TCR T cell receptor Cas9 CRISPR associated protein 9 [0020 ] The present invention provides an inhibitory chi [0031 ] said polypeptide sequence comprises at least one meric antigen receptor (N -CAR ) comprising : sequence from a Tumor -necrosis - factor related apoptosis [0021 ] an extracellular domain comprising an antigen inducing ligand ( TRAIL ) receptor or at least one sequence binding domain ; from a CD200 receptor 1 . [ 0022 ] a transmembrane domain ; [0032 ] The present invention provides a N -CAR accord [0023 ] an intracellular domain ; ing to the above , comprising a polypeptide sequence from a [0024 ] wherein said N -CAR comprises a polypeptide Tumor -necrosis - factor related apoptosis inducing ligand sequence involved in inducing an inhibitory transduction ( TRAIL ) receptor. signal, [0033 ] The present invention provides a N - CAR accord [0025 ] said polypeptide sequence comprises at least one ing to the above comprising at least one polypeptide sequence from a Tumor -necrosis -factor related apoptosis sequence from a polypeptide sequence selected from the list inducing ligand ( TRAIL ) receptor or at least one sequence consisting of SEQ ID NO : 33 (human TR10D ), SEQ ID NO : 34 (human TRIOA ) or SEO ID NO : 35 (human TRIOB ) and from a CD200 receptor 1 . a fragment thereof. [0026 ] The present invention provides an inhibitory chi [0034 ] The present invention provides a N -CAR accord meric antigen receptor (N -CAR ) comprising : ing to any one of the above embodiments , wherein said [0027 ] an extracellular domain comprising an antigen polypeptide sequence has more than 80 % , preferably 90 % binding domain ; and more preferably 95 % identity with a sequence from [0028 ] a transmembrane domain ; SEQ ID NO : 33 , SEQ ID NO : 34 or SEQ ID NO : 35 or a [0029 ] an intracellular domain ; fragment thereof. [0030 ] wherein said N -CAR comprises a polypeptide [0035 ] The present invention provides a N - CAR accord sequence involved in inducing an inhibitory transduction ing to any one of the above embodiments , comprising at signal upon binding of said extracellular domain comprising least one of the following polypeptide sequences : amino an antigen binding domain to an antigen and/ or resulting in acids No 181 - 386 from SEQ ID NO : 33 ( human TR10D ) , a decrease in CTL activity , amino acids No 230 -468 from SEQ ID NO : 34 (human US 2017 /0296623 A1 Oct . 19 , 2017 TR10A ) or of amino acids N° 179 -440 from SEQ ID NO : 35 [0053 ] The N -CAR according to any one of above (human TR10B ) , or a fragment thereof. embodiments , wherein said antigen binding domain binds to [0036 ] The present invention provides a N -CAR accord CD19 , CD20 , CD22 , BCMA , PSMA , CD56 , CD205 , CD83 , ing to any one of the above embodiments wherein said CD206 , CD200 or CD36 . antigen binding domain binds to a cell surface antigen N , [0054 ] The N -CAR according to any one of above and N being not expressed on a cancerous cell and N being embodiments , wherein said antigen binding domain binds to expressed on a non -cancerous cell or a healthy cell. troponin C , beta - 1 integrin , CCKBR , GALR1 or CUBN . 10037 ] The present invention provides a N -CAR accord [0055 ] The N -CAR according to any one of above ing to any one of the above embodiments , wherein the embodiments , wherein said antigen binding domain binds to antigen binding domain binds to a cell - surface antigen N , N CD4 , CD20 , CD22 , CD25 or MUC1. being present in normal tissue but not present or present at 10056 ) The N - CAR according to any one of above undetectable level on a tumor as determined by FACS or embodiments , wherein said antigen binding domain binds to western blot analysis or by any appropriate technique allow troponin C , beta - 1 integrin , CCKBR , GALR1 or MUC1. ing proteins to be quantified . [0057 ] The N -CAR according to any one of above [ 0038 ] In the present invention non - cancerous cell or a embodiments wherein the transmembrane domain com healthy cell also expressing a P antigen , said prises the transmembrane region of PD - 1 . [ 0039 ] P antigen being also expressed or over expressed 10058 ] The N -CAR according to any one of above on a cancerous cell. embodiments wherein the N -CAR comprises the transmem [0040 ] The present invention provides a N - CAR accord brane region of PD - 1 , or a fragment thereof. ing to any one of the above embodiments wherein said [0059 ] The N - CAR according to any one of above antigen binding domain binds to at least one cell surface embodiments wherein the transmembrane domain com antigen N selected from CD56 , CD205, CD83 , CD206 , prises the transmembrane region (s ) of CD8 alpha . CD200 , CD36 , troponin C , beta - 1 integrin , CCKBR , [ 0060 ] The N - CAR according to any one of the above GALR1 CUBN , CD4, CD20 , CD22, CD25 , MUC1, CD19, embodiments wherein the transmembrane domain is BCMA , and PSMA . attached to the extracellular domain of the N -CAR via a [ 0041] The present invention also provides hinge. (0042 ] The N -CAR according to the above embodiments comprising at least one polypeptide sequence consisting [0061 ] The N -CAR according to the above embodiments essentially of amino acids No 181- 386 from SEQ ID NO : 33 wherein the hinge is a human immunoglobulin hinge . (human TR10D ) . [0062 ] The N -CAR according to the above embodiments [ 0043] The N -CAR according to any one of the above wherein the hinge is an IgG1 hinge or a CD8 alpha hinge . embodiments , comprising at least one polypeptide sequence [0063 ] The N -CAR according to any one of the above consisting essentially of amino acids No 230 - 468 from SEQ embodiments wherein the transmembrane domain com ID NO : 34 (human TR10A ). prises a transmembrane region ( s ) of the alpha , beta or zeta [0044 ] The N - CAR according to any one of the above chain of the T - cell receptor, PD - 1 , 4 - 1BB , OX40 , ICOS , embodiments , comprising at least one polypeptide sequence CTLA - 4 , LAG3, 2B4 , BTLA4 , TIM - 3 , TIGIT, SIRPA , consisting essentially of amino acids No 179 -440 from SEQ CD28 , CD3 epsilon , CD45 , CD4 , CD5 , CD8, CD9 , CD16 , ID NO : 35 (human TR10B ) . CD22 , CD33 , CD37 , CD64 , CD80 , CD86 , CD134 , CD137 [0045 ] The N -CAR according comprising at least one or CD154 . polypeptide sequence from a CD200 Receptor 1, preferably [0064 ] The present invention provides a vector encoding a comprising a sequence of SEQ ID NO . 36 or a fragment N -CAR according to any one of the above embodiments. thereof. [0065 ] The present invention provides an immune cell, [0046 ] An N -CAR according to any one of the above preferably a primary immune T cell comprising a P -CAR embodiments, wherein said N -CAR is a single chain ( sc ) N . comprising: CAR or a multi -chain (mc ) N -CAR . 100661 an extracellular domain comprising an antigen [0047 ] The N -CAR according to any one of the above binding domain ; embodiments comprising at least one polypeptide sequence [0067 ] a transmembrane domain ; encoded by a sequence selected from the list consisting in [0068 ] an intracellular domain ; preferably a intracellu SEQ ID NO . 102 to SEQ ID NO . 212 . lar domain comprising an activator transducing domain [0048 ] The N - CAR according to any one of the above and an co -stimulatory domain , embodiments wherein the antigen binding domain binds to [0069 ] and a N -CAR according to any one of the above an off - tissue antigen . embodiments . [ 0049 ] An N -CAR according to the above embodiments , [0070 ] In a preferred embodiment activation of the wherein said extracellular domain comprises at least one a N -CAR inhibits the signal transduction activity related to the single chain variable fragment scFv . P -CAR , resulting in particular in a decrease in the CTL [0050 ] The N - CAR according to any one of the above activity or the immune cell bearing a N -CAR and a P -CAR . embodiments, wherein said antigen binding domain com [ 0071 ] In one preferred embodiment the present invention prises a Fv, a Fab , or a ( Fab ' ) 2 . provides a immune cell according to the above embodiment [ 0051] The N -CAR according to any one of the above wherein at least one gene encoding a TCR alpha or a TCR embodiments wherein said antigen binding domain binds to beta subunit is inactivated , preferably by deletion using a a cell surface antigen N , expressed on a non cancerous cell specific endonuclease . or a healthy cell expressing a P antigen . [0072 ] In one embodiment the present invention provides [ 0052 ] In one embodiment N is P , in a preferred embodi an immune cell according to any one of the above embodi ment N is not P. ments wherein at least one gene encoding a TCR and a gene US 2017 /0296623 A1 Oct . 19 , 2017 encoding a deoxicitidine kinase ( dck ) are inactivated , pref- binding domain of the P -CAR binds to is CD123 and the erably by deletion using a endonuclease, preferably a antigen to which the antigen binding domain of the N - CAR TALEN . binds is CD4, CD20 , CD22 , CD25 or MUC1. [0073 ] In one embodiment the present invention provides [0089 ] In one embodiment the present invention provides an immune cell according to any one of the above embodi an immune cell according to any one of the above embodi ments for use as a medicament. ments wherein the cell surface antigen to which the antigen [0074 ] In one embodiment the present invention provides binding domain of the P -CAR binds to is ROR1 and the cell an immune cell according to any one of the above embodi surface antigen to which the antigen binding domain of the ments for use in the prevention or treatment of a haemato N -CAR binds is troponin C , beta - 1 integrin , CCKBR , logical cancer condition , preferably a relapsed refractory GALR1 or MUC1. haematological cancer. [0090 ] In one embodiment the present invention provides 10075 ]. The immune cell according to the above embodi an immune cell for use as a medicament ments, wherein said haematological cancer condition is [0091 ] In one embodiment the present invention provides leukemia or myeloma, preferably relapsed and / or refractory an immune cell for the treatment of a leukemia selected from leukemia or relapsed and / or refractory myeloma . the group consisting of acute myelogenous leukemia [0076 ] In one embodiment the present invention provides (AML ). a method of engineering an immune cell according to any [0092 ] The present invention provides an immune cell one of the above embodiments comprising : according to the above embodiments for use in therapy , [ 0077 ] ( a ) Providing an immune cell ; optionally delet wherein the condition is a pre -malignant or malignant cancer ing a candidate gene , said candidate gene being pref condition characterized by CD123 - expressing cells or by erably TCRA and dCK CLL - 1 expressing cells [0078 ] (b ) Expressing a N -CAR and a P - CAR according [0093 ) The immune cell according to any one of the above to the invention at the cell surface . [0079 ] (c ) optionally deleting a candidate gene, said embodiments , wherein said haematological cancer condition candidate gene being preferably selected from TCRA , is multiple myeloma (MM ) . PD1, CTLA4 and dCK (0094 ] The immune cell according to the above embodi [0080 ] In one embodiment the present invention provides ments for use in therapy , wherein the condition is a pre a method as above wherein said immune cells are provided malignant or malignant cancer condition characterized by from a donor, preferably a healthy donor. CD38 - expressing cells . [0081 ] In one embodiment the present invention provides [0095 ] The immune cell according to the above embodi a vector comprising a sequence selected from the list con ments , wherein said haematological cancer condition is sisting in SEQ ID NO . 102 to SEQ ID NO . 212 . chronic lymphocytic leukemia (CLL ) . [ 0082 ] The immune cell according to the above embodi [0096 ] The immune cell according to the above embodi ments is provided wherein the immune cell is a T -cell , ments for use in therapy , wherein the condition is a pre preferably a CD4 T cell or a CD8 T cell , more preferably a malignant or malignant cancer condition characterized by primary CD8 T cell. CS1 - expressing cells or by ROR1 - expressing cells . [0083 ] The immune cell according to the above embodi [0097 ] The immune cell according to the above embodi ments wherein the T- cell is a human T- cell, preferably a ments for use in therapy , wherein the condition is a solid primary immune T cell . tumor such as breast , colon , lung , or kidney tumor charac [0084 ] The immune cell according to the above embodi terized especially by ROR1 - expressing cells . ments wherein the P CAR binds to at least one of the 10098 ] The immune cell according to the above embodi following antigen CD123 , CD22 , CS1, CD38 , CD19 , CD33 , ments for use in therapy , wherein the condition is a pre CD20 . malignant or malignant cancer condition characterized by [0085 ] The immune cell according to the above embodi CD22 -expressing cells . ments is selected from primary inflammatory T- lympho [0099 ] The immune cell according to any one of the above cytes , primary cytotoxic T -lymphocytes , primary regulatory wherein the reduction of activation of the immune cells T - lymphocytes or primary helper T- lymphocytes. when both the P -CAR and N -CAR bind to their respective [0086 ] In one embodiment the present invention provides antigens is increased , preferably by at least 5 % , 10 % , 15 % , an immune cell according to any one of the above embodi 20 % or 30 % as compared to the same immune cell wherein ments , wherein the cell surface antigen to which the antigen a P -CAR alone binds to its cell surface antigen , and /or as binding domain of the P -CAR binds to is CD38 and the cell compared to an immune cell expressing a full intracellular surface antigen to which the antigen binding domain of the domain of PD - 1 or a full intracellular domain of CTLA - 4 as N -CAR binds to is CD56 , CD205 , CD83 , CD206 , CD200 or an intracellular domain of said N -CAR . CD36 . (0100 ] The immune cell according to any one of the above [0087 ] In one embodiment the present invention provides embodiments wherein the level of activation of the immune an immune cell according to any one of the above embodi cell is determined by measuring cytokine production . ments , wherein the cell surface antigen to which the antigen 10101 ] The immune cell according to claim 34 wherein the binding domain of the P - CAR binds to is CS1 and the cell cytokine is IFNgamma or TNFalpha . surface antigen to which the antigen binding domain of the [0102 ] The immune cell according to any one of the above N -CAR binds to is troponin C , beta - 1 integrin , CCKBR , embodiments wherein the cytokine production is measured GALR1 or CUBN . by ELISA and / or FACS and /or luminex . [0088 ] In one embodiment the present invention provides [0103 ] The immune cell according to any one of the above an immune cell according to any one of the above embodi - embodiments wherein the level of activation of the immune ments , wherein the cell surface antigen to which the antigen cell is determined by the level of degranulation . US 2017 /0296623 A1 Oct . 19 , 2017 [0104 ] The immune cell according to any one of the above surface expression below the level of detection using any embodiments wherein degranulation is measured by mea appropriate technique such as flow cytometry analysis , suring expression of CD107a by FACS . western blot or Elisa test or that said polypeptide is [ 0105 ] The immune cell according to any one of the above expressed at a level of less than 20 % and preferably less than embodiments wherein the level of activation of the immune 10 % and more preferably at undetectable level in a given cell is measured by monitoring the ability of the immune cell cell as compared the expression of said polypeptide mea to kill target cells . sured in a cell known to express said polypeptide used as a [0106 ] The present invention provides a method of engi positive control. neering an immune cell according to any one of the above [0124 ] This can be tested by currently used techniques embodiments comprising : allowing expression of proteins to be measured and quan [0107 ] ( a ) Introducing into said cell at least one poly tified that is using western blot, flow cytometry analysis , nucleotide encoding the N -CAR and at least one poly Elisa test and others . nucleotide encoding the CAR ; [0125 ] According to one embodiment, the intracellular [0108 ] (b ) Expressing said polynucleotides into said domain of the N - CAR comprises a polypeptide sequence cell . involved in transduction signal of a receptor, and said 10109 ] The present invention provides a method for treat receptor is not significantly expressed in T -cells . ing a patient in need thereof comprising : [0126 ] In a preferred embodiment, said polypeptide [0110 ] a ) Providing an immune cell according to any sequence is from a sequence selected from the group con one of the above embodiments, and; sisting of SEQ ID NO : 1 (human KI2L2 ) , SEO ID NO : 2 [0111 ] b ) Administrating said T -cells to said patient. (human KI2L1) , SEQ ID NO : 3 human FCG2B ) , SEQ ID [0112 ] The present invention discloses a method for treat NO : 4 (human KI2L3 ) , SEQ ID NO : 5 (human KI3L2 ) , SEO ing a patient according to any one of the above embodiments ID NO : 6 ( human KI2L4) , SEQ ID NO : 7 (human KI3L1) , wherein said immune cells are recovered from patients . SEQ ID NO : 8 (human KIZLA ) , SEQ ID NO : 9 (human [0113 ] The different objects of the present invention are MILR1) , SEO ID NO : 10 (human LIRB4 ) , SEO ID NO : 11 disclosed in details as follows (human LIRB3 ), SEQ ID NO : 12 (human KI3L3 ), SEQ ID [0114 ] Inhibitory Chimeric Antigen Receptor ( iCAR or NO : 15 (human LIRB5) , SEQ ID NO : 16 ( human LIRB2 ), N - CAR ) SEQ ID NO : 18 (human FCRL4 ) , SEQ ID NO :23 (human [0115 ] The present invention relates to an inhibitory chi FCRL5 ) , SEQ ID NO : 24 (human FCRL2) , SEQ ID NO : 25 meric antigen receptor (iCAR or N -CAR ) comprising : (human FCRL1) , SEQ ID NO : 27 (human FCRL3) , SEO ID [0116 ] an extracellular domain comprising an antigen NO :28 (human MPZL1) , SEQ ID NO : 29 (human PILRA ) , binding domain ; SEO ID NO :30 ( human PVR ) , and a fragment thereof. [0117 ] a transmembrane domain ; [0127 ] In a more preferred embodiment said polypeptide 0118 ] an intracellular domain ; sequences is SEQ ID NO : 1 (human KI2L2 ) or a fragment [ 0119 ] wherein said intracellular domain comprises a thereof. polypeptide sequence involved in transduction signal said [0128 ] In a more preferred embodiment said polypeptide polypeptide sequence is not significantly expressed in T- cell , sequences is SEQ ID NO : 2 (human KI2L1) , or a fragment preferably in primary T cells , and / or said polypeptide thereof. sequence is from a Tumor- necrosis - factor related apoptosis [0129 ] In a more preferred embodiment said polypeptide inducing ligand ( TRAIL ) receptor and / or CD200 receptor 1 , sequences is SEQ ID NO : 3 (human FCG2B ), or a fragment [ 0120 ] provided said polypeptide sequence not expressed thereof. in T cells is not an ITMS, preferably it does not consists in [0130 ] In a more preferred embodiment said polypeptide (or does not comprise ) a sequence chosen in a group sequences is SEQ ID NO : 4 (human KI2L3 ) , or a fragment consisting of SEQ ID NO : 13 ( human SIGL8 ), SEQ ID thereof. NO : 14 (human SIGL7 ) , SEQ ID NO : 17 ( human SIGL5 ) , 10131 ] In a more preferred embodiment said polypeptide SEQ ID NO : 20 (human SIGL9 ) , SEQ ID NO : 21 (human sequences is SEQ ID NO : 5 (human KI3L2 ) , or a fragment SIGL6 ) , SEO ID NO : 22 ( human CD33 ) , SEQ ID NO : 26 thereof. (human SIG12 ), SEQ ID NO :31 (human SIG11 ) , SEQ ID [0132 ] In a more preferred embodiment said polypeptide NO : 32 (human SIG10 ) and SEQ ID NO : 19 (human sequences is SEQ ID NO :6 (human KI2L4 ) , or a fragment PECA1) . thereof. [ 0121 ] Intracellular Domain of the CAR - N According to [0133 ] In a more preferred embodiment said polypeptide the Invention sequences is SEQ ID NO :7 (human KI3L1) , or a fragment [0122 ] According to one embodiment, the intracellular thereof. domain of the N -CAR comprises a polypeptide sequence [0134 ] In a more preferred embodiment said polypeptide from a receptor involved in transduction signal which is not sequences is SEQ ID NO : 8 ( human KIZLA ) , or a fragment significantly expressed in non - engineered T - cells . By “ not thereof. significantly expressed ” is meant that the protein involved in [0135 ]. In a more preferred embodiment said polypeptide the transduction signal, which intracellular domain is from , sequences is SEQ ID NO : 9 ( human MILR1) , or a fragment is not expressed or expressed at a significant lower level in thereof. the same culture or growth conditions in a non - engineered [0136 ] In a more preferred embodiment said polypeptide T -Cell . By " engineered T - cells ” are meant T - cells that have sequences is SEQ ID NO : 10 (human LIRB4) , or a fragment been genetically modified to express or to unable expression thereof of a given genetic sequence . [0137 ] In a more preferred embodiment said polypeptide [ 0123 ] Here , by “ not significantly expressed ” , it is meant sequences is SEQ ID NO : 11 (human LIRB3) , or a fragment that the expression of said polypeptide, preferably cell thereof. US 2017 /0296623 A1 Oct . 19 , 2017 [ 0138 ] In a more preferred embodiment said polypeptide [0147 ] In a more preferred embodiment said polypeptide sequences is SEQ ID NO : 12 ( human KI3L3) , or a fragment sequences is SEQ ID NO : 29 ( human PILRA ) , or a fragment thereof. thereof . [ 0139 ] In a more preferred embodiment said polypeptide [0148 ] In a more preferred embodiment said polypeptide sequences is SEQ ID NO : 15 (human LIRB5 ) , or a fragment sequences is SEQ ID NO : 30 (human PVR ), or a fragment thereof. [0140 ] In a more preferred embodiment said polypeptide thereof . sequences is SEQ ID NO : 16 (human LIRB2) , or a fragment [0149 ] In another embodiment according to the invention , thereof. said polypeptide sequence has more than 80 % , preferably [ 0141 ] In a more preferred embodiment said polypeptide 90 % and more preferably 95 % identity with a sequence from sequences is SEQ ID NO : 18 (human FCRL4 ) , or a fragment a sequence selected from SEQ ID NO : 1 (human KI2L2 ) , thereof. SEQ ID NO : 2 (human KI2L1) , SEQ ID NO : 3 (human [0142 ] In a more preferred embodiment said polypeptide FCG2B ) , SEQ ID NO : 4 (human KI2L3 ) , SEQ ID NO : 5 sequences is SEQ ID NO :23 (human FCRL5 ), or a fragment (human KI3L2 ), SEQ ID NO : 6 (human KI2L4 ) , SEQ ID thereof. NO : 7 (human KI3L1) , SEQ ID NO : 8 ( human KIZLA ) , SEQ [ 0143 ] In a more preferred embodiment said polypeptide ID NO : 9 (human MILR1) , SEQ ID NO : 10 (human LIRB4) , sequences is SEQ ID NO : 24 (human FCRL2) , or a fragment SEQ ID NO : 11 (human LIRB3) , SEQ ID NO : 12 (human thereof. KI3L3 ) , SEO ID NO : 15 (human LIRB5 ) , SEO ID NO : 16 [ 0144 ] In a more preferred embodiment said polypeptide ( human LIRB2) , SEQ ID NO : 18 (human FCRL4 ), SEQ ID sequences is SEQ ID NO : 25 (human FCRL1) , or a fragment NO : 23 (human FCRL5) , SEQ ID NO :24 ( human FCRL2 ), thereof. SEO ID NO : 25 (human FCRL1) , SEQ ID NO : 27 (human [0145 ] In a more preferred embodiment said polypeptide FCRL3 ), SEQ ID NO : 28 ( human MPZL1 ), SEQ ID NO :29 sequences is SEQ ID NO : 27 (human FCRL3 ) , or a fragment (human PILRA ), SEQ ID NO :30 (human PVR ). thereof. [0150 ] The above sequences are presented in the following [0146 ] In a more preferred embodiment said polypeptide Table 1 . sequences is SEQ ID NO : 28 (human MPZL1 ) , or a fragment [ 0151 ] The architecture of some exemplary N -CARs is thereof. presented in the following Table 2 . TABLE 1 Amino -acid sequences of intracellular domain from a receptor involved in transduction signal which is not significantly expressed in non - engineered T - cell ( Sequences SEO NO : 13 (human SIGL8) , SEQ ID NO : 14 (human SIGL7 ) , SEQ ID NO : 17 (human SIGL5 ) , SEQ ID NO : 20 (human SIGL9 ) , SEQ ID NO : 21 (human SIGL6 ) , SEQ ID NO : 22 ( human CD33 ) , SEQ ID NO : 26 ( human SIG12 ) , SEQ ID NO : 31 (human SIG11 ) , SEQ ID NO : 32 ( human SIG10 ) and SEQ ID NO : 19 being not part of the present invention ) UniProt SEQ Name ID ID NO : human KI2L2 P43627 1 MWSHLNRLLFWSIFSSVTCRKAVLDCEAMKTNEFPSPCLDSKTKVVMKGONVSMFCSHKNKSLQITYSLFRRK THLGTODGKGEPAIFNLSITEAHESGPYKCKAOVTSCSKYSRDFSFTIVDPVTSPVLNIMIOTETDRHITLH CLSVNGSLPINYTFFENHVAISPAISKYDREPAEFNLTKKNPGEEEEYRCEAKNRLPNYATYSHPVTMPSTGG DSCPFCLKLLLPGLLLLLVIILILAFWVLPKYKTRKAMRNNVPRDRGDTAMEVGIYANILEKOAKEESVPEV GSRPCVSTAQDEAKHSQELQYATPVFQEVAPREQEACDSYKSGYVYSELNF human KI2L1 P43626 MIPTFTALLCLGLSLGPRTHMQAGPLPKPTLWAEPGSVISWGNSVTIWCQGTLEAREYRLDKEESPAPWDRON PLEPKNKARFSIPSMTEDYAGRYRCYYRSPVGWSOPSDPLELVMTGAYSKPTLSALPSPLVTSGKSVTLLCOS RSPMDTFLLIKERAAHPLLHLRSEHGAQOHOAEFPMSPVTSVHGGTYRCFSSHGFSHYLLSHPSDPLELIVSG SLEDPRPSPTRSVSTAAGPEDOPLMPTGSVPHSGLRRHWEVLIGVLVVSILLLSLLLFLLLOHWROGKHRTLA OROADFORPPGAAEPEPKDGGLORRSSPAADVOGENFCAAVKNTOPEDGVEMDTROSPHDEDPOAVTYAKVKH SRPRREMASPPSPLSGEFLDTKDRQAEEDROMDTEAAASEAPODVTYAQLHSFTLROKATEPPPSQEGASPAE PSVYATLAIH human FCG2B P31994 MTPALTALLCLGLSLGPRTRVOAGPFPKPTLWAEPGSVISWGSPVTIWCOGSOEAQEYRLHKEGSPEPLDRNN PLEPKNKARFSIPSMTEHHAGRYRCHYYSSAGWSEPSDPLEMVMTGAYSKPTLSALPSPVASGGNMTLRCGS QKGYHHFVLMKEGEHOLPRTLDSQQLHSRGFQALFPVGPVTPSHRWRFTCYYYYTNTPWVWSHPSDPLEILPS GVSRKPSLLTLQGPVLAPGQSLTLQCGSDVGYNRFVLYKEGERDFLQRPGOQPQAGLSQANFTLGPVSPSNGG QYRCYGAHNLSSEWSAPSDPLNILMAGQIYDTVSLSAQPGPTVASGENVTLLCQSWWQFDTFLLTKEGAAHPP LRLRSMYGAHKYQAEFPMSPVTSAHAGTYRCYGSYSSNPHLLSHPSEPLELVVSGHSGGSSLPPTGPPSTPGL GRYLEVLIGVSVAFVLLLFLLLFLLLRRORHSKHRTSDORKTDFORPAGAAETEPKDRGLLRRSSPAADVOEE NLYAAVKDTQSEDRVELDSQSPHDEDPQAVTYAPVKHSSPRREMASPPSSLSGEFLDTKDROVEEDROMDTEA AASEASQDVTYAQLHSLTLRRKATEPPPSQEGEPPAEPSIYATLAIH human KI2L3 P43628 MSLMVVSMACVGFFLLEGPWPHVGGODKPFLSAWPGTWVSEGQHVTLQCRSRLGFNEFSLSKEDGMPVPELYN RIFRNSFLMGPVTPAHAGTYRCCSSHPHSPTGWSAPSNPVVIMVTGVHRKPSLLAHPGPLVKSGETVILQCWS DVRFERFLLHREGITEDPLRLVGQLHDAGSOVNYSMGPMTPALAGTYRCFGSVTHLPYELSAPSDPLDIVWG LYGKPSLSAQPGPTVQAGENVTLSCSSRSLFDIYHLSREAEAGELRLTAVLRVNGTFQANFPLGPVTHGGNYR CFGSFRALPHAWSDPSDPLPVSVTGNSRHLHVLIGTSVVIIPFAILLFFLLHRWCANKKNAVVMDQEPAGNRT VNREDSDEQDPQEVTYAQLNHCVFTQRKITRPSQRPKTPPTDTSV US 2017 /0296623 A1 Oct . 19, 2017 TABLE 1 - continued Amino - acid sequences of intracellular domain from a receptor involved in transduction signal which is not significantly expressed in non - engineered T - cell ( Sequences SEQ NO : 13 (human SIGL8 ) , SEQ ID NO : 14 (human SIGL7 ) , SEQ ID NO : 17 (human SIGL5 ) , SEQ ID NO : 20 (human SIGL9 ) , SEQ ID NO : 21 ( human SIGL6 ) , SEQ ID NO : 22 (human CD33 ) , SEQ ID NO : 26 ( human SIG12 ) , SEQ ID NO : 31 (human SIG11) , SEQ ID NO : 32 (human SIG10 ) and SEQ ID NO : 19 being not part of the present invention ) UniProt SEO Name 1IDID ID NO : human KI3L2 P43630 55 MLLLLLLLPLLWGTKGMEGDRQYGDGYLLQVQELVTVQEGLCVHVPCSFSYPQDGWTDSDPVHGYWFRAGDRP YQDAPVATNNPDREVQAETQGRFQLLGDIWSNDCSLSIRDARKRDKGSYFFRLERGSMKWSYKSQLNYKTKQL SVFVTALTHRPDILILGTLESGHSRNLTCSVPWACKOGTPPMISWIGASVSSPGPTTARSSVLTLTPKPODHG TSLTCOVTLPGTGVTTTSTVRLDVSYPPWNLTMTVFOGDATASTALGNGSSLSVLEGOSLRLVCAVNSNPPAR LSWTRGSLTLCPSRSSNPGLLEL PRVHVRDEGEFTCRAQNAQGSQHISLSLSLQNEGTGTSRPVSQVTLAAVG GAGATALAFLSFCIIFIIVRSCRKKSARPAAGVGDTGMEDAKAIRGSASQGPLTESWKDGNPLKKPPPAVAPS SGEEGELHYATLSFHKVKPQDPQGQEATDSEYSEIKIHKRETAETQACLRNHNPSSKEVRG human KI2L4 099706 6 MLLLLLLPLLWGRERVEGQKSNRKDYSLTMQSSVTVQEGMCVHVRCSFSYPVDSQTDSDPVHGYWFRAGNDIS WKAPVATNNPAWAVOEETRDRFHLLGDPOTKNCTLSIRDARMSDAGRYFFRMEKGNIKWNYKYDOLSVNVTAL THRPNILIPGTLESGCFONLTCSVPWACEOGTPPMI SWMGTSVSPLHPSTTRSSVLTLIPOPOHHGTSLTCOV TLPGAGVTTNRTIOLNVSYPPONLTVTVFOGEGTASTALGNSSSLSVLEGOSLRLVCAVDSNPPARLSWTWRS LTLYPSOP SNPLVLELOVHLGDEGEFTCRAONSLGSOHVSLNLSLOOEYTGKMRPVSGVLLGAVGGAGATALV FLSFCVIFIVVRSCRKKSARPAADVGDIGMKDANTIRGSASQGNLTESWADDNPRHHGLAAHSSGEEREIQYA PLSFHKGEPODLSGQEATNNEYSEIKIPK human KI3L1 P43629 MTLTLSVLICLGLSVGPRTCVQAGTLPKPTLWAEPASVIARGKPVTLWCQGPLETEEYRLDKEGLPWARKRON PLEPGAKAKFHIPSTVYDSAGRYRCYYETPAGWSEPSDPLELVATGFYAEPTLLALPSPVVASGGNVTLOCDT LDGLLTFVLVEEEOKLPRTLYSOKLPKGPSQALFPVGPVTPSCRWRFRCYYYYRKNPQVWSNPSDLLEILVPG VSRKPSLLIPOGSVARGGSLTLQCRSDVGYDIFVLYKEGEHDLVQGSGQQPQAGLSQANFTLGPVSRSHGGQ YRCYGAHNLSPRWSAPSDPLDILIAGLIPDIPALSVOPGPKVASGENVTLLCOSWHQIDTFFLTKEGAAHPPL CLKSKYQSYRHQAEFSMSPVTSAQGGTYRCYSAIRSYPYLLSSPSYPQELVVSGPSGDPSLSPTGSTPTPGPE DOPLTPTGLDPQSGLGRHLGVVTGVSVAFVLLLFLLLFLLLRHRHQS KHRTSAHFYRPAGAAGPEPKDQGLOK RASPVADIQEEILNAAVKDTQPKDGVEMDARAAASEAPODVTYAQLHSLTLRREATEPPPSQEREPPAEPSIY APLAIH human KI2LA Q8N109 8 MTPIVTVLICLGLSLGPRTHVQTGTIPKPTLWAEPDSVITOGSPVTLSCOGSLEAQEYRLYREKKSASWITRI RPELVKNGOFHIPSI TWEHTGRYGCOYYSRARWSELSDPLVLVMTGAYPKPTLSAOPSPVVTSGGRVTLOCES QVAFGGFILCKEGEE EHPQCLNSOPHARGSSRAIFSVGPVSPNRRWSHRCYGYDLNSPYVWSSPSDLLELLVP GVSKKPSLSVQPGPVVAPGESLTLQCVSDVGYDRFVLYKEGERDLRQLPGRQPQAGLSQANFTLGPVSRSYGG QYRCYGAHNLSSECSAPSDPLDILI TGOIRGTPFISVOPGPTVASGENVTLLCOSWROFHTFLLTKAGAADAP LRLRSIHEYPKYQAEFPMSPVTSAHAGTYRCYGSLNSDPYLLSHPSEPLELVVSGPSMGSSPPPTGPISTPAG PEDQPLTPTGSDPQSGLGRHLGVVIGILVAWLLLLLLLLLFLILRHRROGKHWTSTORKADFQHPAGAVGPE PTDRGLQWRSSPAADAQEENLYAAVKDTOPEDGVEMDTRAAASEAPODVTYAQLHSLTLRRKATEPPPSQERE PPAEPSIYATLAIH human MILR1 Q7Z6M3 MLPLLLLPLLWGGSLOEKPVYELOVOKSVTVOEGLCVLVPCSFSYPWRSWYSSPPLYVYWFRDGEIPYYAEVV ATNNPDRRVKPETQGRFRLLGDVOKKNCSLSIGDARMEDTGSYFFRVERGRDVKYSYQONKLNLEVTALIEKP DIHFLEPLESGRPTRLSCSLPGSCEAGPPLTFSWTGNALSPLDPETTRSSELTLTPRPEDHGTNLTCOMKROG AOVTTERTVOLNVSYAPOTITIFRNGIALEILONTSYLPVLEGOALRLLCDAPSNPPAHLSWFOGSPALNATP ISNTGILELRRVRSAEEGGFTCRAOHPLGFLOIFLNLSVYSLPOLLGPSCSWEAEGLHCRCSFRARPAPSLCW RLEEKPLEGNSSQGSFKVNSSSAGPWANSSLILHGGLSSDLKVSCKAWNIYGSQSGSVLLLOGRSNLGTGVVP AALGGAGVMALLCICLCLIFFLIVKARRKQAAGRPEKMDDEDPIMGTITSGSRKKPWPDSPGDQASPPGDAPP LEEQKELHYASLSFSEMKSREPKDQEAPSTTEYSEIKTSK human LIRB 4 O8NHJ6 10 MLLWASLLAFAPVCGOSAAAHKPVISVHPPWTTFFKGERVTLTCNGFOFYATEKTTWYHRHYWGEKLTLTPGN TLEVRESGLYRCQARGSPRSNPVRLLFSSDSLILQAPYSVFEGDTLVLRCHRRRKEKLTAVKYTWNGNILSIS NKSWDLLIPQASSNNNGNYRCIGYGDENDVFRSNFKIIKIQELFPHPELKATDSOPTEGNSVNLSCETOLPPE RSDTPLHFNFFRDGEVILSDWSTYPELOLPTVWRENSGSYWCGAETVRGNIHKHSPSLOIHVORIPVSGVLLE TOPSGGQAVEGEMLVLVCSVAEGTGDTTFSWHREDMOESLGRKTORSLRAELELPAIROSHAGGYYCTADNSY GPVQSMVLNVTVRETPGNRDGLVAAGATGGLLSALLLAVALLFHCWRRRKSGVGFLGDETRLPPAPGPGESSH SICPAQVELQSLYVDVHPKKGDLVYSEIQTTOLGEEEEANTSRTLLEDKDVSVVYSEVKTQHPDNSAGKISSK DEES human LIRB3 075022 11 MOPRWAOGATMWLGVLLTLLLCSSLEGOENSFTINSVDMKSLPDWTVONGKNLTLOCFADVSTTSHVKPOHOM LFYKDDVLFYNISSMKSTESYFIPEVRIYDSGTYKCTVIVNNKEKTTAEYQLLVEGVPSPRVTLDKKEAIQGG IVRVNCSVPEEKAPIHFTIEKLELNEKMVKLKREKNSRDONFVILEFPVEEQDRVLSFRCQARIISGIHMQTS ESTKSELVTVTESFSTPKFHISPTGMIMEGAQLHIKCTIQVTHLAQEFPEIIIQKDKAIVAHNRHGNKAVYSV MAMVEHSGNYTCKVESSRISKVSSIVNITELFSKPELESSFTHLDOGERLNLSCSIPGAPPANFTIOKEDTI VSQTQDFTKIASKSDSGTYICTAGIDKVVKKSNTVQIVVCEMLSOPRISYDAQFEVIKGQTIEVRCESISGTL PISYQLLKTSKVLENSTKNSNDPAVFKDNPTEDVEYQCVADNCHSHAKMLSEVLRVKVIAPVDEVQISILSSK VVESGEDIVLOCAVNEGSGPITYKFYREKEGKPFYOMTSNATOAFWTKOKASKEOEGEYYCTAFNRANHASSV PRSKILIVRVILAPWKKGLIAVVIIGVIIALLIIAAKCYFLRKAKAKOMPVEMSRPAVPLLNSNNEKMSDPNM EANSHYGHNDDVRNHAMKPINDNKEPLNSDVQYTEVQVSSAESHKDLGKKDTETVYSEVRKAVPDAVESRYSR TEGSLDGT US 2017 /0296623 A1 Oct . 19, 2017 10 TABLE 1 - continued Amino - acid sequences of intracellular domain from a receptor involved in transduction signal which is not significantly expressed in non - engineered T - cell ( Sequences SEQ NO : 13 (human SIGL8 ) , SEQ ID NO : 14 (human SIGL7 ) , SEQ ID NO : 17 (human SIGL5 ) , SEQ ID NO : 20 (human SIGL9 ) , SEQ ID NO : 21 ( human SIGL6 ) , SEQ ID NO : 22 (human CD33 ) , SEQ ID NO : 26 ( human SIG12 ) , SEQ ID NO : 31 (human SIG11) , SEQ ID NO : 32 (human SIG10 ) and SEQ ID NO : 19 being not part of the present invention ) UniProt SEO Name ID ID NO : human KI3L3Q8N743 12 MLLLLLPLLWGRERAEGOTSKLL TMOSSVTVOEGLCVHVPCSFSYPSHGWIYPGPVVHGYWFREGANTDODAP VATNNPARAVWEETRDRFHLLGDPHTKNCTLSIRDARRSDAGRYFFRMEKGSIKWNYKHHRLSVNVTAL THRP NILIPGTLESGCPONLTCSVPWACEOGTPPMI SWIGTSVSPLDPSTTRSSVLTLIPOPODHGTSLTCOVTFPG ASVTTNKTVHLNVSYPPONLTMTVFOGDGTVSTVLGNGSSLSLPEGOSLRLVCAVDAVDSNPPARLSLSWRGL TLCPSQPSNPGVLELPWVHLRDAAEFTCRAQNPLGSQQVYLNVSLOSKATSGVTQGVVGGAGATALVFLSFCV IFVVVRSCRKKSARPAAGVGDTGIEDANAVRGSASOGPLTEPWAEDSPPDOPPPASARSSVGEGELOYASLSF QMVKPWDSRGQEATDTEYSEIKI HR human SIGL8 O9NYZ4 13 MOGAQEASASEMLPLLLPLLWAGALAQERRFOLEGPESLTVOEGLCVLVPCRLPTTLPASYYGYGYWFLEGAD VPVATNDPDEEVOEETRGRFHLLWDPRRKNCSLSIRDARRRDNAAYFFRLKSKWMKYGYTSSKLSVRVMALTH RPNISIPGTLESGHPSNLTCSVPWVCEOGTPPIFSWMSAAPTSLGPRTTOSSVLTITPRPODHSTNLTCOVTF PGAGVTMERTIOLNVSYAPOKVAISIFOGNSAAFKILONTSSLPVLEGOALRLLCDADGNPPAHLSWFOGFPA LNATPISNTGVLELPOVGSAEEGDFTCRAOHPLGSLOISLSLFVHWKPEGRAGGVLGAVWGASITTLVFLCVC FIFRVKTRRKKAAQPVQNTDDVNPVMVSGSRGHQHQFQTGIVSDHPAEAGPISEDEQELHYAVLHFHKVQPQE PKVTDTEYSEIKIHK human SIGL7 Q9Y286 1414 MPLLLLLPLLWAGALAMDPNFWLOVOESVTVOEGLCVLVPCTFFHPIPYYDKNSPVHGYWFREGAIISRDSPV ATNKLDQEVQEETQGRFRLLGDPSRNNCSLSIVDARRRDNGSYFFRMERGSTKYSYKSPQLSVHVTDLTHRPK ILIPGTLEPGHSKNLTCSVSWACEQGTPPIFSWLSAAPTSLGPRTTHSSVLIITPRPQDHGTNLTCOVKFAGA GVTTERTIQLNVTYVPONPUGIFPGDGSGKGETRAGVVHGAIGGAGVTALLALCLCLIFFIVKTHRRKAARTA VGRNDTHPTTGSASPKHQKKSKLHGPTETSSCSGAAPTVEMDEELHYASLNFHGMNPSKDTSTEYSEVRTO human LIRB5 075023 15 MLLWVILLVLAPVSGQFARTPRPIIFLOPPWTTVFQGERVTLTCKGFRFYSPOKTKWYHRYLGKEILRETPDN ILEVQESGEYRCQAQGSPLSSPVHLDFSSASLILQAPLSVFEGDSVVLRCRAKAEVTLNNTIYKNDNVLAFLN KRTDFHIPHACLKDNGAYRCTGYKESCCPVSSNTVKIOVOEPFTRPVLRASSFOPISGNPVTLTCETOLSLER SDVPLRFRFFRDDOTLGLGWSLSPNFQITAMWSKDSGFYWCKAATMPYSVISDSPRSWIQVQIPASHPVLTLS PEKALNFEGTKVTLHCETOEDSLRTLYRFYHEGVPLRHKSVRCERGASISFSLTTENSGNYYCTADNGLGAKP SKAVSLSVTVPVSHPVLNLSSPEDLIFEGAKVTLHCEAORGSLPILYOFHHEGAALERRSANSAGGVAISFSL TAEHSGNYYCTADNGFGPORSKAVSLSVTVPVSHPVLTLSSAEALTFEGATVTLHCEVORGSPOILYOFYHED MPLWSSSTPSVGRVSFSFSLTEGHSGNYYCTADNGFGPORSEVVSLFVTVPVSRPILTLRVPRAOAVVGDLLE LHCEAPRGSPPILYWFYHEDVTLGSSSAPSGGEASFNLSLTAEHSGNYSCEANNGLVAOHSDTISLSVIVPVS RPILTFRAPRAQAVVGDLLELHCEALRGSSPILYWFYHEDVTLGKISAPSGGGASFNLSLTTEHSGIYSCEAD NGLEAQRSEMVTLKVAVPVSRPVLTLRAPGTHAAVGDLLELHCEALRGSPLILYRFFHEDVTLGNRSSPSGGA SLNLSLTAEHSGNYSCEADNGLGAQRSETVTLYITGLTANRSGPFATGVAGGLLSIAGLAAALLLYCWLSRK AGRKPASDPARSPSDSDSQEPTYHNVPAWEELQPVYTNANPRGENVVYSEVRIIQEKKKHAVASDPRHLRNKG SPIIYSEVKVASTPVSGSLFLAS SAPHR human LIRB2 Q8N423 16 MLLWSLLVIFDAVTEOADSLTLVAPSSVFEGDSIVLKCOGEONWKIOKMAYHKDNKELSVFKKFSDFLIOSAV LSDSGNYFCSTKGQLFLWDKTSNIVKIKVQELFQRPVLTASSFQPIEGGPVSLKCETRLSPORLDVQLQFCFF RENOVLGSGWSSSPELOISAVWS EDTGSYWCKAETVTHRIRKOSLOSOIHVORIPISNVSLEIRAPGGOVTEG OKLILLCSVAGGTGNVTFSWYREATGTSMGKKTORSLSAELEIPAVKESDAGKYYCRADNGHVPIOSKVYNIP VRIPVSRPVLTLRSPGAOAAVGDLLELHCEALRGSPPILYOFYHEDVTLGNSSAPSGGGASFNLSLTAEHSGN YSCEANNGLGAOCSEAVPVSISGPDGYRRDLMTAGVLWGLFGVLGFTGVALLLYALFHKISGESSATNEPRGA SRPNPQEFTYSSPTPDMEELQPVYVNVGSVDVDVVYSQVWSMQQPESSANIRTLLENKDSQVIYSSVKKS human SIGL5 015389 17 MLPRLLLLICAPLCEPAELFLIASPSHPTEGSPVTLTCKMPFLOSSDAQFOFCFFRDTRALGPGWSSSPKLOI AAMWKEDTGSYWCEAOTMASKVLRSRRSOINVHRVPVADVSLETOPPGGOVMEGDRLVLICSVAMGTGDITFL WYKGAVGLNLOSKTORSLTAEYEIPSVRESDAEOYYCVAENGYGPSPSGLVSITVRIPVSRPILMLRAPRAOA AVEDVLELHCEALRGSPPILYWFYHEDITLGSRSAPSGGGASFNLSLTEEHSGNYSCEANNGLGAQRSEAVTL NFTVPTGARSNHLTSGVIEGLLSTLGPATVALLFCYGLKRKIGRRSARDPLRSLPSPLPQEFTYLNSPTPGOL QPIYENVNVVSGDEVYSLAYYNOPEQESVAAETLGTHMEDKVSLDIYSRLRKANITDVDYEDAM human FCRL4 096PJ5 18 MLLLLLLLPPLLCGRVGAKEQKDYLLTMQKSVTVQEGLCVSVLCSFSYPONGWTASDPVHGYWFRAGDHVSRN IPVATNNPARAVOEETRDRFHLLGDPONKDCTLSIRDTRESDAGTYVFCVERGNMKWNYKYDOLSVNVTASOD LLSRYRLEVPESVTVOEGLCVSVPCSVLYPHYNWTASSPVYGSWFKEGADIPWDIPVATNTPSGKVOEDTHGR FLLLGD POTNNCSLSIRDARKGDSGKYYFOVERGSRKWNYIYDKLSVHVTALTHMPTFSIPGTLESGHPRNLT PTITRSSMLSLIPOPODHGTSLTCOVTLPGAGVTMTRAVRLNISYPP QNLTMTVFQGDGTASTTLRNGSALSVLEGQSLHLVCAVDSNPPARLSWTWGSLTLSPSQSSNLGVLELPRVHV KDEGEFTCRAQNPLGSQHISLSLSLQNEYTGKMRPISGVTLGAFGGAGATALVFLYFCIIFVVVRSCRKKSAR PAVGVGDTGMEDANAVRGSASOGPLIESPADDSPPHHAPPALATPSPEEGEIQYASLSFHKARPOYPOEOEAI GYEYSEINIPK human PECA1 P16284 19 MLLWLLLLILTPGREQSGVAPKAVLLLNPPWSTAFKGEKVALICSSISHSLAQGDTYWYHDEKLLKI KHDKIO ITEPGNYOCKTRGSSLSDAVHVEFSPDWLILOALHPVFEGDNVILRCOGKDNKNTHOKVYYKDGKOLPNSYNL EKITVNSVSRDNSKYHCTAYRKFYILDIEVTSKPLNIQVQELFLHPVLRASSSTPIEGSPMTLTCETQLSPOR PDVQLQFSLFRDSQTLGLGWSRS PRLQIPAMWTEDSGSYWCEVETVTHSIKKRSLRSQIRVORVPVSNVNLEI RPTGGQLIEGENMVLICSVAQGSGTVTFSWHKEGRVRSLGRKTORSLLAELHVLTVKESDAGRYYCAADNVHS US 2017 /0296623 A1 Oct . 19, 2017 TABLE 1 - continued Amino - acid sequences of intracellular domain from a receptor involved in transduction signal which is not significantly expressed in non - engineered T - cell ( Sequences SEQ NO : 13 (human SIGL8 ) , SEQ ID NO : 14 (human SIGL7 ) , SEQ ID NO : 17 (human SIGL5 ) , SEQ ID NO : 20 (human SIGL9 ) , SEQ ID NO : 21 ( human SIGL6 ) , SEQ ID NO : 22 (human CD33 ) , SEQ ID NO : 26 ( human SIG12 ) , SEQ ID NO : 31 (human SIG11) , SEQ ID NO : 32 (human SIG10 ) and SEQ ID NO : 19 being not part of the present invention ) UniProt SEO Name IDID ID NO : PILSTWIRVTVRIPVSHPVLTFRAPRAHTVVGDLLELHCESLRGSPPILYRFYHEDVTLGNSSAPSGGGASFN LSLTAEHSGNYSCDADNGLGAOHSHGVSLRVTVPVSRPVLTLRAPGAOAVVGDLLELHCESLRGSFPILYWFY HEDDTLGNISAHSGGGASFNLSLTTEHSGNYSCEADNGLGAQHSKVVTLNVTGTSRNRTGLTAAGITGLVLSI LVLAAAAALLHYARARRKPGGLSATGTSSHSPSECOEPSSSRPSRIDPOEP THSKPLAPMELEPMYSNVNPGD SNPIYSQIWSIQHTKENSANCPMMHQEHEELTVLYSELKKTHPDDSAGEASSRGRAHEEDDEENYENVPRVLL ASDH human SIGL9 Q9Y336 20 MAASAGAGAVIAAPDSRRWLWSVLAAALGLLTAGVSALEVYTPKEIFVANGTOGKLTCKFKSTSTTGGLTSVS WSFQPEGADTTVSFFHYSQGQVYLGNYPPFKDRI SWAGDLDKKDASINIENMQFIHNGTYICDVKNPPDIVVO PGHIRLYVVEKENLPVFPVWVWVGIVTAVLGLTLLISMILAVLYRRKNSKRDYTGCSTSESLSPVKQAPRKS PSDTEGLVKSLPSGSHQGPVIYAQLDHSGGHHSDKINKSESVVYADIRKN human SIGL6 043699 21 MGRPLLLPLLPLLLPPAFLOPSGSTGSGPSYLYGVTOP KHLSASMGGSVEIPFSFYYPWELATAPDVRISWRR GHFHROSFYSTRPPSIHKDYVNRLFLNWTEGOKSGFLRISNLOKODOSVYFCRVELDTRSSGROOWOSIEGTK LSITOAVTTTTORPSSMTTTWRLSSTTTTTGLRVTOGKRRSDSWHISLETAVGVAVAVTVLGIMILGLICLLR WRRRKGQQRTKATTPAREPFONTEEPYENIRNEGONTDPKLNPKDDGIVYASLALSSSTSPRAPPSHRPLKSP ONETLYSVLKA human CD33 P20138 22 MARAMAAAWPLLLVALLVLSWPPPGTGDVVVOAPTOVPGFLGDSVTLPCYLOVPNMEVTHVSOL TWARHGESG SMAVFHQTQGPSYSESKRLEFVAARLGAELRNASLRMFGLRVEDEGNYTCLFVTFPCGSRSVDIWLRVLAKPQ NTAEVQKVOLTGEPVPMARCVSTGGRPPAQITWHSDLGGMPNTSQVPGFLSGTVTVTSLWILVPSSQVDGKNV TCKVEHESFEKPQLLTVNLTVYYPPEVSISGYDNNWYLGQNEATLTCDARSNPEPTGYNWSTTMGPLPPFAVA QGAQLLIRPVDKPINTTLICNVTNALGARQAELTVOVKEGPPSEHSGISRNAIIFLVLGILVFLILLGIGIYF YWSKCSREVLWHCHLCPSSTEHASASANGHVSYSAVSRENSSSQDPQTEGTR human FCRL5 096RD9 23 MVPGQAQPQS PEMLLLPLLLPVLGAGSLNKDPSYSLQVQRQVPVPEGLCVIVSCNLSYPRDGWDESTAAYGYW FKGRTSPKTGAPVATNNOSREVEMSTRDRFOLTGDPGKGSCSLVIRDAOREDEAWYFFRVERGSRVRHSFLSN AFFLKVTALTKKPDVYIPETLEPGOPVTVICVFNWAFKKCPAPSFSWTGAALSPRRTRPSTSHFSVLSFTPSP ODHDTDLTCHVDFSRKGVSAORTVRLRVAYAPKDLIISISHDNTSALELOGNVIYLEVOKGOFLRLLCAADSO PPATLSWVLQDRVLSSSHPWGPRTLGLELRGVRAGDSGRYTCRAENRLGSQQQALDLSVQYPPENLRVMVSQA NRTVLENLGNGTSLPVLEGOSLRLVCVTHSSPPARLSWTRWGOTVGPSOPSDPGVLELPPIOMEHEGEFTCHA OHPLGSOHVSLSLSVHYPPOLLGPSCSWEAEGLHCSCSSOASPAPSLRWWLGEELEGNSSOGSFEVTPSSAGP WANSSLSLHGGLSSGLRLRCKAWNVHGAQSGSVFQLLPGKLEHGGGLGLGAALGAGVAALLAFCSCLWFRVK ICRKEARKRAAAEQDVPSTLGPISQGHQHECSAGSSQDHPPPGAATYTPGKGEEQELHYASLSFOGLRLWEPA DQEAPSTTEYSEIKIHTGOPLRGPGFGLQLEREMSGMVPK human FCRL2 096LA5 24 MLLPLLLSSLLGGSQAMDGRFWIRVQESVMVPEGLCISVPCSFSYPRODWTGSTPAYGYWFKAVTETTKGAPV ATNHOSREVEMS TRGRFOLTGDPAKGNCSLVIRDAQMODESOYFFRVERGSYVRYNFMNDGFFLKVTAL TOKP DVYIPETLEPGOPVTVICVFNWAFEECPPPSFSWTGAALSSOGTKPTTSHFSVLSFTPRPODHNTDLTCHVDF SRKGVSAQRTVRLRVAYAPRDLVISISRDNTPALEPOPOGNVPYLEAOKGOFLRLLCAADSOPPATLSWVLON RVLSSSHPWGPRPLGLELPGVKAGDSGRYTCRAENRLGSOORALDLSVOYPPENLRVMVSOANRTVLENLGNG TSLPVLEGOSLCLVCVTHSSPPARLSWTORGOVLSPSOPSDPGVLELPRVOVEHEGEFTCHARHPLGSOHVSL SLSVHYSPKLLGPSCSWEAEGLHCSCSSOASPAPSLRWWLGEELLEGNSSODSFEVTPSSAGPWANSSLSLHG GLSSGLRLRCEAWNVHGAQSGSILQLPDKKGLISTAFSNGAFLGIGITALLFLCLALIIMKILPKRRTOTETP RPRFSRHSTILDYINVPTAGPLAQKRNOKATPNSPRTPLPPGAPSPESKKNQKKQYQLPSFPEPKSSTQAPE SQESQEELHYATLNFPGVRPRPEARMPKGTQADYAEVKFO human FCRL1 096LA6 25 MWSHLNRLLFWSIFSSVTCRKAVLDCEAMKTNEFPSPCLDSKTKVVMKGONVSMFCSHKNKSLOITYSLFRRK THLGTQDGKGEPAIFNLSITEAHESGPYKCKAQVTSCSKYSRDFSFTIVDPVTSPVLNIMVIQTETDRHITLH CLSVNGSLPINYTFFENHVAISPAISKYDREPAEFNLTKKNPGEEEEYRCEAKNRLPNYATYSHPVTMPSTGG DSCPFCLKLLLPGLLLLLVVIILILAFWVLPKYKTRKAMRNNVPRDRGDTAMEVGIYANILEKQAKEESVPEV GSRPCVSTAQDEAKHSQELQYATPVFQEVAPREQEACDSYKSGYVYSELNF human SIG12 096P01 26 MIPTFTALLCLGLSLGPRTHMOAGPLPKPTLWAEPGSVISWGNSVTIWCOGTLEAREYRLDKEESPAPWDRON PLEPKNKARFSIPSMTEDYAGRYRCYYRSPVGWSOPSDPLELVMTGAYSKPTLSALPSPLVTSGKSVTLLCOS RSPMDTFLLIKERAAHPLLHLRSEHGAQOHOAEFPMSPVTSVHGGTYRCFSSHGFSHYLLSHPSDPLELIVSG SLEDPRPSPTRSVSTAAGPEDOPLMPTGSVPHSGLRRHWEVLIGVLVVSILLLSLLLFLLLOHWROGKHRTLA QRQADFQRPPGAAEPEPKDGGLQRRSSPAADVQGENFCAAVENTOPEDGVEMDTROSPHDEDPQAVTYAKVKH SRPRREMASPPSPLSGEFLDTKDROAEEDROMDTEAAASEAPODVTYAOLHSFTLROKATEPPPSOEGASPAE PSVYATLAIH human FCRL3 096P31 27 MTPALTALLCLGLSLGPRTRVQAGPFPKPTLWAEPGSVISWGSPVTIWCOGSQEAQEYRLHKEGSPEPLDRNN PLEPKNKARFSIPSMTEHHAGRYRCHYYSSAGWSEPSDPLEMVMTGAYSKPTLSALPSPVVASGGNMTLRCGS OKGYHHFVLMKEGEHOLPRTLDSOOLHSRGFOALFPVGPVTPSHRWRFTCYYYYTNTPWVWSHPSDPLEILPS GVSRKPSLLTLQGPVLAPGQSLTLQCGSDVGYNRFVLYKEGERDFLORPGQQPQAGLSQANFTLGPVSPSNGG QYRCYGAHNLSSEWSAPSDPLNILMAGQIYDTVSLSAQPGPTVASGENVTLLCQSWWQFDTFLLTKEGAAHPP LRLRSMYGAHKYQAEFPMSPVTSAHAGTYRCYGSYSSNPHLLSHPSEPLELVVSGHSGGSSLPPTGPPSTPGL US 2017 /0296623 A1 Oct . 19, 2017 12 TABLE 1 - continued Amino - acid sequences of intracellular domain from a receptor involved in transduction signal which is not significantly expressed in non - engineered T - cell ( Sequences SEQ NO : 13 (human SIGL8 ) , SEQ ID NO : 14 (human SIGL7 ) , SEQ ID NO : 17 (human SIGL5 ) , SEQ ID NO : 20 (human SIGL9 ) , SEQ ID NO : 21 ( human SIGL6 ) , SEQ ID NO : 22 (human CD33 ) , SEQ ID NO : 26 ( human SIG12 ) , SEQ ID NO : 31 (human SIG11) , SEQ ID NO : 32 (human SIG10 ) and SEQ ID NO : 19 being not part of the present invention ) UniProt SEO Name IDID ID NO : GRYLEVLIGVSVAFVLLLFLLLFLLLRRORHSKHRTSDORKTDFORPAGAAETEPKDRGLLRRSSPAADVOEE NLYAAVKDTQSEDRVELDSQSPHDEDPQAVTYAPVKHSSPRREMASPPSSLSGEFLDTKDROVEEDROMDTEA AASEASQDVTYAQLHSLTLRRKATEPPPSQEGEPPAEPSIYATLAIH human MP ZL1 095297 28 MSLMVVSMACVGFFLLEGPWPHVGGQDKPFLSAWPGTVVSEGQHVTLQCRSRLGFNEFSLSKEDGMPVPELYN RIFRNSFLMGPVTPAHAGTYRCCSSHPHSPTGWSAPSNPVVIMVTGVHRKPSLLAHPGPLVKSGETVILOCWS DVRFERFLLHREGITEDPLRLVGQLHDAGSQVNYSMGPMTPALAGTYRCFGSVTHLPYELSAPSDPLDIVVVG LYGKPSLSAQPGPTVQAGENVTLSCSSRSLFDIYHLSREAEAGELRL TAVLRVNGTFQANFPLGPVTHGGNYR CFGSFRALPHAWSDPSDPLPVSVTGNSRHLHVLIGTSVVIIPFAILLFFLLHRWCANKKNAVVMDOEPAGNRT VNREDSDEQDPQEVTYAQLNHCVFTORKITRPSQRPKTPPTDTSV humanhuman PILRAPILRA O9UKJ1Q9UKJ1 29 MLLLLLLLPLLWGTKGMEGDRQYGDGYLLQVQELVTVQEGLCVHVPCSFSYPODGWTDSDPVHGYWFRAGDRPMLLLLLLLPLLWGTKGM YODAPVATNNPDREVOAETOGRFOLLGDIWSNDCSLSIRDARKRDKGSYFFRLERGSMKWSYKSOLNYKTKOL SVFVTALTHRPDILILGTLESGHSRNLTCSVPWACKOGTPPMI SWIGASVSSPGPTTARSSVLTLTPKPODHG TSLTCQVTLPGTGVTTTSTVRLDVSYPPWNLTMTVFQGDATASTALGNGSSLSVLEGQSLRLVCAVNSNPPAR LSWTRGSLTLCPSRSSNPGLLEL PRVHVRDEGEFTCRAONAOGSOHISLSLSLONEGTGTSRPVSOVTLAAVG GAGATALAFLSFCIIFIIVRSCRKKSARPAAGVGDTGMEDAKAIRGSASOGPLTESWKDGNPLKKPP PAVAPS SGEEGELHYATLSFHKVKPQDPQGQEATDSEYSEIKIHKRETAETQACLRNHNPSSKEVRG human PVR P15151 30 MLLLLLLPLLWGRERVEGOKSNRKDYSLTMOSSVTVQEGMCVHVRCSFSYPVDSQTDSDPVHGYWFRAGNDIS WKAPVATNNPAWAVQEETRDRFHLLGDPQTKNCTLSIRDARMSDAGRYFFRMEKGNIKWNYKYDOLSVNVTAL THRPNILIPGTLESGCFQNLTCSVPWACEQGTPPMI SWMGTSVSPLHPSTTRSSVLTLIPQPQHHGTSLTCOV TLPGAGVTTNRTIOLNVSYPPONLTVTVFOGEGTASTALGNSSSLSVLEGOSLRLVCAVDSNPPARLSWTWRS LTLYPSQPSNPLVLELQVHLGDEGEFTCRAONSLGSQHVSLNLSLQQEYTGKMRPVSGVLLGAVGGAGATALV FLSFCVIFIVVRSCRKKSARPAADVGDIGMKDANTIRGSASQGNLTESWADDNPRHHGLAAHSSGEEREIQYA PLSFHKGEPODLSGOEATNNEYSEIKIPK human SIG11 096RL6 31 MTLTLSVLICLGLSVGPRTCVOAGTLPKPTLWAEPASVIARGKPVTLWCOGPLETEEYRLDKEGLPWARKRON PLEPGAKAKFHIPSTVYDSAGRYRCYYETPAGWSEPSDPLELVATGFYAEPTLLALPSPVVASGGNVTLOCDT LDGLLTFVLVEEEOKLPRTLYSOKLPKGPSOALFPVGPVTPSCRWRFRCYYYYRKNPOVWSNPSDLLEILVPG VSRKPSLLIPOGSVVARGGSLTLOCRSDVGYDIFVLYKEGEHDLVOGSGOOPOAGLSOANFTLGPVSRSHGGO YRCYGAHNLSPRWSAPSDPLDILIAGLIPDIPALSVQPGPKVASGENVTLLCOSWHQIDTFFLTKEGAAHPPL CLKSKYQSYRHQAEFSMSPVTSAQGGTYRCYSAIRSYPYLLSSPSYPQELVVSGPSGDPSLSPTGSTPTPGPE DOPLTPTGLDPOSGLGRHLGVVTGVSVAFVLLLFLLLFLLLRHRHQS KHRTSAHFYRPAGAAGPEPKDQGLOK RASPVADIQEEILNAAVKDTQPKDGVEMDARAAASEAPODVTYAQLHSLTLRREATEPPPSQEREPPAEPSIY APLAIH human SIG10 096LC7 32 MTPIVTVLICLGLSLGPRTHVQTGTIPKPTLWAEPDSVITQGSPVTLSCQGSLEAQEYRLYREKKSASWITRI RPELVKNGQFHIPSI TWEHTGRYGCQYYSRARWSELSDPLVLVMTGAYPKPTLSAQPSPVVTSGGRVTLQCES QVAFGGFILCKEGEEEHPQCLNS QPHARGSSRAIFSVGPVSPNRRWSHRCYGYDLNSPYVWSSPSDLLELLVP GVSKKPSLSVQPGPVVAPGESLTLQCVSDVGYDRFVLYKEGERDLRQLPGRQPQAGLSQANFTLGPVSRSYGG QYRCYGAHNLSSECSAPSDPLDILI TGQIRGTPFISVQPGPTVASGENVTLLCQSWRQFHTFLLTKAGAADAP LRLRSIHEYPKYQAEFPMS PVTSAHAGTYRCYGSLNSDPYLLSHPSEPLELVVSGPSMGSSPPPTGPISTPAG PEDOPLTPTGSDPOSGLGRHLGVVIGILVAVVLLLLLLLLLFLILRHRROGKHWTSTORKADFOHPAGAVGPE PTDRGLQWRSSPAADAQEENLYAAVKDTQPEDGVEMDTRAAASEAPODVTYAQLHSLTLRRKATEPPPSQERE PPAEPSIYATLAIH TABLE 2 Example of N - CARs according to the invention Plasmid encoding Amino acids of the N - CAR signal peptide GS linker 1 VH chain GS linker VL chain GS linker 2 inhibitory polypeptide used pCLS27446 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 229 - 325 of SEQ ID NO . 36 ( SEQ ID NO . 100 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27447 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 230 -468 of SEQ ID NO . 34 ( SEQ ID NO . 101) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27448 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 179 -440 of SEQ ID NO . 35 ( SEQ ID NO . 102 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27449 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 181 - 386 of SEQ ID NO . 33 (SEQ ID NO . 103) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27450 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 229 - 364 of SEQ ID NO . 22 (SEQ ID NO . 104 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 US 2017 /0296623 A1 Oct . 19, 2017 TABLE 2 -continued Example of N - CARs according to the invention Plasmid encoding Amino acids of the N - CAR signal peptide GS linker 1 VH chain GS linker VL chain GS linker 2 inhibitory polypeptide used PCLS27451 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 214 - 310 of SEQ ID NO . 3 ( SEQ ID NO . 105) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 pCLS27452 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 292 -429 of SEQ ID NO . 25 ( SEQ ID NO . 106 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 pCLS27453 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 388- 508 of SEQ ID NO . 24 (SEQ ID NO . 107) NO . 37 NO . 39 NO . 41 NO . 38 NO . 4 NO . 40 PCLS27454 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 564 -734 of SEQ ID NO . 27 ( SEQ ID NO . 108 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27455 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 375- 515 of SEQ ID NO . 18 ( SEQ ID NO . 109 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27456 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 753 -977 of SEQ ID NO . 23 (SEQ ID NO . 110 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27457 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 420 -598 of SEQ ID NO . 16 ( SEQ ID NO . 111) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27445 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 420 -631 of SEQ ID NO . 11 (SEQ ID NO . 112 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27458 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 219 -448 of SEQ ID NO . 10 ( SEQ ID NO . 113 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27459 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 419 -590 of SEQ ID NO . 15 ( SEQ ID NO . 114 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27460 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 214 - 343 of SEQ ID NO . 9 ( SEQ ID NO . 115) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27461 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 147- 269 of SEQ ID NO . 28 ( SEQ ID NO . 116 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27462 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 151- 303 of SEQ ID NO . 29 ( SEQ ID NO . 117 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 pCLS27464 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 329- 417 of SEQ ID NO . 30 (SEQ ID NO . 118 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27465 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 442 -697 of SEQ ID NO . 32 ( SEQ ID NO . 119 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27466 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 453- 698 of SEQ ID NO . 31 ( SEQ ID NO . 120 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27467 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 463 -595 of SEQ ID NO . 26 ( SEQ ID NO . 121) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27468 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 331 -551 of SEQ ID NO . 17 ( SEQ ID NO . 122 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27469 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 334 -453 of SEQ ID NO . 21 ( SEQ ID NO . 123 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27470 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 337 - 467 of SEQ ID NO . 14 (SEQ ID NO . 124 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27471 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 345- 499 of SEQ ID NO . 13 (SEQ ID NO . 125 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27472 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 237- 463 of SEQ ID NO . 20 ( SEQ ID NO . 126 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27473 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 206 - 348 of SEQ ID NO . 2 (SEQ ID NO . 127) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27474 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 206 -348 of SEQ ID NO . 1 ( SEQ ID NO . 128 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27475 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 206 - 341 of SEQ ID NO . 4 (SEQ ID NO . 129 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27476 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 203- 377 of SEQ ID NO . 6 (SEQ ID NO . 130 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27477 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 201 - 375 of SEQ ID NO . 8 ( SEQ ID NO . 131) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 pCLS27478 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 301 - 444 of SEQ ID NO . 7 ( SEQ ID NO . 132 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27479 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 301- 455 of SEQ ID NO . 5 (SEQ ID NO . 133 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 pCLS27480 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 296 -410 of SEQ ID NO . 12 (SEQ ID NO . 134 ) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27481 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 592 -738 of SEQ ID NO . 19 (SEQ ID NO . 135) NO . 37 NO . 39 NO . 41 NO . 38 NO . 42 NO . 40 PCLS27482 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 229- 325 of SEQ ID NO . 36 ( SEQ ID NO . 138 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27483 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 230 -468 of SEQ ID NO . 34 (SEQ ID NO . 139 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27484 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 179 - 440 of SEQ ID NO . 35 ( SEQ ID NO . 140 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 pCLS27485 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 181 - 386 of SEQ ID NO . 33 ( SEQ ID NO . 141) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27486 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 229 - 364 of SEQ ID NO . 22 (SEQ ID NO . 142 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 US 2017 /0296623 A1 Oct . 19, 2017 14 TABLE 2 -continued Example of N - CARs according to the invention Plasmid encoding Amino acids of the N -CAR signal peptide GS linker 1 VH chain GS linker VL chain GS linker 2 inhibitory polypeptide used pCLS27487 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 214 - 310 of SEQ ID NO . 3 ( SEQ ID NO . 143 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 pCLS27488 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 292 -429 of SEQ ID NO . 25 ( SEQ ID NO . 144 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 pCLS27489 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 388- 508 of SEQ ID NO . 24 (SEQ ID NO . 145 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27490 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 564 -734 of SEQ ID NO . 27 ( SEQ ID NO . 146 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27491 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 375- 515 of SEQ ID NO . 18 ( SEQ ID NO . 147) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27492 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 753 -977 of SEQ ID NO . 23 (SEQ ID NO . 148 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27493 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 420 -598 of SEQ ID NO . 16 (SEQ ID NO . 149 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27494 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 420 -631 of SEQ ID NO . 11 (SEQ ID NO . 150 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27495 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 219 -448 of SEQ ID NO . 10 (SEQ ID NO . 151) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27496 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 419 -590 of SEQ ID NO . 15 (SEQ ID NO . 152 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27497 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 214 - 343 of SEQ ID NO . 9 (SEQ ID NO . 153) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27498 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 147- 269 of SEQ ID NO . 28 ( SEQ ID NO . 154 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 pCLS27499 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 151- 303 of SEQ ID NO . 29 ( SEQ ID NO . 155 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 pCLS27501 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 329- 417 of SEQ ID NO . 30 ( SEQ ID NO . 156 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27502 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 442 -697 of SEQ ID NO . 32 ( SEQ ID NO . 157 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27503 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 453 -698 of SEQ ID NO . 31 ( SEQ ID NO . 158 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27504 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 463 -595 of SEQ ID NO . 26 (SEQ ID NO . 159 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 pCLS27505 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 331 -551 of SEQ ID NO . 17 ( SEQ ID NO . 160 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27506 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 334 -453 of SEQ ID NO . 21 ( SEQ ID NO . 161) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27507 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 337 - 467 of SEQ ID NO . 14 (SEQ ID NO . 162) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27508 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 345- 499 of SEQ ID NO . 13 (SEQ ID NO . 163) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27509 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 237- 463 of SEQ ID NO . 20 ( SEQ ID NO . 164 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 pCLS27510 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 206 - 348 of SEQ ID NO . 2 (SEQ ID NO . 165) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27511 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 206 -348 of SEQ ID NO . 1 ( SEQ ID NO . 166 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 pCLS27512 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 206 - 341 of SEQ ID NO . 4 (SEQ ID NO . 167 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 pCLS27513 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 203- 377 of SEQ ID NO . 6 ( SEQ ID NO . 168 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27514 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 201 - 375 of SEQ ID NO . 8 (SEQ ID NO . 169) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 pCLS27515 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 301 - 444 of SEQ ID NO . 7 ( SEQ ID NO . 170 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27516 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 301- 455 of SEQ ID NO . 5 ( SEQ ID NO . 171 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27517 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 296 -410 of SEQ ID NO . 12 ( SEQ ID NO . 172 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27518 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 592 -738 of SEQ ID NO . 19 ( SEQ ID NO . 173 ) NO . 37 NO . 39 NO . 43 NO . 38 NO . 44 NO . 40 PCLS27698 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 229- 325 of SEQ ID NO . 36 ( SEQ ID NO . 175 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 PCLS27699 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 230 -468 of SEQ ID NO . 34 (SEQ ID NO . 176 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 PCLS27700 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 179 - 440 of SEQ ID NO . 35 ( SEQ ID NO . 177 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 PCLS27701 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 181 - 386 of SEQ ID NO . 33 (SEQ ID NO . 178 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 PCLS27702 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 229 - 364 of SEQ ID NO . 22 (SEQ ID NO . 179) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 US 2017 /0296623 A1 Oct . 19, 2017 TABLE 2 -continued Example of N - CARs according to the invention Plasmid encoding Amino acids of the N -CAR signal peptide GS linker 1 VH chain GS linker VL chain GS linker 2 inhibitory polypeptide used pCLS27703 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 214 - 310 of SEQ ID NO . 3 ( SEQ ID NO . 180 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 PCLS27704 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 292 -429 of SEQ ID NO . 25 (SEQ ID NO . 181 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 pCLS27705 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 388 -508 of SEQ ID NO . 24 (SEQ ID NO . 182 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 PCLS27706 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 564 -734 of SEQ ID NO . 27 ( SEQ ID NO . 183 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 PCLS27707 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 375- 515 of SEQ ID NO . 18 ( SEQ ID NO . 184 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 PCLS27708 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 753 -977 of SEQ ID NO . 23 ( SEQ ID NO . 185 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 PCLS27709 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 420 -598 of SEQ ID NO . 16 ( SEQ ID NO . 186 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 PCLS27710 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 420 -631 of SEQ ID NO . 11 (SEQ ID NO . 187) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 pCLS27711 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 219 -448 of SEQ ID NO . 10 ( SEQ ID NO . 188 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 PCLS27712 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 419 - 590 of SEQ ID NO . 15 (SEQ ID NO . 189 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 PCLS27713 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 214 - 343 of SEQ ID NO . 9 (SEQ ID NO . 190 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 PCLS27714 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 147- 269 of SEQ ID NO . 28 ( SEQ ID NO . 191) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 PCLS27715 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 151- 303 of SEQ ID NO . 29 ( SEQ ID NO . 192 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 pCLS27716 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 329- 417 of SEQ ID NO . 30 ( SEQ ID NO . 193 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 PCLS27717 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 442 -697 of SEQ ID NO . 32 ( SEQ ID NO . 194 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 PCLS27718 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 453 -698 of SEQ ID NO . 31 ( SEQ ID NO . 195 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 pCLS27719 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 463 -595 of SEQ ID NO . 26 ( SEQ ID NO . 196 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 pCLS27720 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 331 -551 of SEQ ID NO . 17 ( SEQ ID NO . 197 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 pCLS27721 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 334 -453 of SEQ ID NO . 21 ( SEQ ID NO . 198 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 PCLS27722 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 337 - 467 of SEQ ID NO . 14 ( SEQ ID NO . 199) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 PCLS27723 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 345- 499 of SEQ ID NO . 13 ( SEQ ID NO . 200 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 PCLS27724 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 237- 463 of SEQ ID NO . 20 (SEQ ID NO . 201) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 pCLS27725 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 206 - 348 of SEQ ID NO . 2 ( SEQ ID NO . 202 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 pCLS27726 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 206 -348 of SEQ ID NO . 1 ( SEQ ID NO . 203) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 pCLS27727 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 206 - 341 of SEQ ID NO . 4 ( SEQ ID NO . 204 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 PCLS27728 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 203- 377 of SEQ ID NO . 6 (SEQ ID NO . 205 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 pCLS27729 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 201 - 375 of SEQ ID NO . 8 ( SEQ ID NO . 206 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 PCLS27730 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 301 -444 of SEQ ID NO . 7 ( SEQ ID NO . 207 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 PCLS27731 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 301- 455 of SEQ ID NO . 5 (SEQ ID NO . 208 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 pCLS27732 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 296 -410 of SEQ ID NO . 12 ( SEQ ID NO . 209 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 pCLS27733 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID 592 -738 of SEQ ID NO . 19 (SEQ ID NO . 210 ) NO . 37 NO . 39 NO . 45 NO . 38 NO . 46 NO . 40 [ 0152 ] According to one embodiment, the N -CAR of the a receptor involved in transduction signal described in the invention comprises at least one of any one of the polypep column “ Amino acids of the inhibitory polypeptide used in tide sequences described in the column “ Amino acids of the table 1 . inhibitory polypeptide used ” in table 1 . [0154 ] According to one embodiment, the intracellular [ 0153 ] According to one embodiment, the inhibitory sig domain of the N -CAR comprises a polypeptide sequence naling transduction domain of the N -CAR of the invention from Tumor -necrosis - factor related apoptosis inducing consists in one of any one of the polypeptide sequences from ligand (TRAIL ) receptor. US 2017 /0296623 A1 Oct . 19 , 2017 16 [ 0155 ] Endogenous TRAIL is expressed as a 281 - amino from a TRAIL receptor, said TRAIL receptor is involved in acid type II trans- membrane protein , which is anchored to in caspase - 8 -mediated apoptosis through proteolytic activa the plasma membrane and presented on the cell surface . TRAIL was independently identified ( Wiley SR , Schooley tion and further NF -kappa - B activation . K Smolak P J , Din W S , Huang CP, Nicholl J K , et al . [0161 ] In one embodiment, the present invention provides “ Identification and characterization of a new member of the an iCAR (or N CAR ) comprising an intracellular signaling TN F family that induces apoptosis . Immunity 1995 ; 3 :673 domain comprising at least one sequence selected from SEQ 82 ; and Pitti R M , Marsters S A , Ruppert S , Donahue O , ID NO : 33 ( human TR10D ), SEQ ID NO : 34 ( human Moore A , Ashkenazi A , 1996 .” Induction of apoptosis by TR10A ) and SEQ ID NO : 35 (human TR10B ) ; According to Apo - 2 ligand , a new member of the tumor necrosis factor one embodiment, the intracellular domain of the N -CAR cytokine family ” . J Biol Chem ; 271: 12687 - 90 . ) In the first consists in one of any one of the polypeptide sequences publication , sequence alignments indicated its close relation selected from SEO ID NO : 33 (human TR10D ) , SEO ID to other death ligands , with highest sequence similarities reported for Fas ligand (FasL ) . NO : 34 (human TR10A ) and SEQ ID NO : 35 ( human [0156 ] TRAIL is expressed by natural killer cells , which , TR10B ); following the establishment of cell - cell contacts , can induce [0162 ] In another embodiment the present invention pro TRAIL - dependent apoptosis in target cells (Smyth M J , vides an iCAR ( or N CAR ) comprising an intracellular Cretney E , Takeda K , Wiltrout R H , Sedger L M , Kayagaki signaling domain comprising at least one sequence having N , et al. , 2001, “ Tumor necrosis factor- related apoptosis more than 80 % , preferably 90 % and more preferably 95 % inducing ligand (TRAIL ) contributes to interferon gamma identity with SEQ ID NO : 33 (human TR10D ) , SEQ ID NO : dependent natural killer cell protection from tumor metas 34 (human TR10A ) and SEQ ID NO : 35 (human TR10B ). tasis . J Exp Med ; 193 :661 - 70 ) . [0163 ] As preferred ones, said polypeptide sequences of [ 0157] Physiologically , the TRAIL - signaling system was receptor have more than 80 % , preferably 90 % and more shown to be essential for immune surveillance , for shaping preferably 95 % identity with SEQ ID NO : 33 (human the immune system through regulating T- helper cell 1 versus TR10D ) , SEQ ID NO : 34 (human TR10A ) and SEQ ID NO : T -helper cell 2 as well as “ helpless” CD8 + T- cell numbers , 35 ( human TR10B ) . Human TRIOD is also called and for the suppression of spontaneous tumor formation TNFRSF10D , DCR2, TRAILR4 or TRUNDD and has as ( Janssen E M , Droin N M , Lemmens E E , Pinkoski M J , ORF names UNQ251/ PRO288 . Human TR10A is also Bensinger Si, Ehst B D , et al. 2005 , “ CD4 + T -cell help called TNFRSF10A , APO2 , DR4 or TRAILR1. Human controls CD8 + T -cell memory via TRAIL -mediated activa TR10B is also called TNFRSF10B , DR5 KILLER , tion - induced cell death ” . Nature ; 434 : 88 - 93 . ) . TRAILR2 , TRICK2 or ZTNFR9 and has as ORF names [ 0158 ] As reviewed in the publication (Hellwig C T , and UNQ160 / PRO186 . Rehm M , 2012 “ TRAIL Signaling and Synergy Mechanisms [0164 ] According to another embodiment, the intracellular Used in TRAIL - Based Combination Therapies” , Mol Can domain of the N -CAR comprises a polypeptide sequence cer Ther. 11 ( 1 ): 3 - 13 ) , TRAIL and agonistic antibodies raised from the CD200 receptor 1 , more preferably a sequence against TRAIL death receptors are highly promising new comprising a sequence of SEQ ID NO : 36 . anticancer agents . In this review is described the recent [0165 ] In another embodiment the present invention pro advances in the molecular understanding of TRAIL signal vides an iCAR (or N CAR ) comprising an intracellular ing and the progress made in using TRAIL or agonistic signaling domain comprising at least one sequence having antibodies clinically in mono - and combination therapies . more than 80 % , preferably 90 % and more preferably 95 % Human agonistic monoclonal antibodies targeting TRAIL identity with SEQ ID NO : 36 . The cell surface glycoprotein R1 (mapatumumab ) or TRAIL -R2 ( lexatumumab ) were CD200 receptor 1 (Uniprot ref: Q8TD46 ) represents another used to treat everal metastatic , triple ( estrogen receptor , example of intracellular domain part of the iCAR (or N progesterone receptor, and HER2 ) - negative cancer cell lines CAR ) of the present invention . This inhibitory receptor for (Malin D , Chen F, Schiller C , Koblinski J, Cryns V L . the CD200 /OX2 cell surface glycoprotein limits inflamma 2011 " Enhanced metastasis suppression by targeting TRAIL tion by inhibiting the expression of proinflammatory mol receptor 2 in a murine model of triple -negative breast ecules including TNF - alpha , interferons , and inducible nitric cancer. ” Clin Cancer Res . 17 ( 15 ) : 5005 - 15 ) . oxide synthase (iNOS ) in response to selected stimuli [0159 ] These publications, a disclose an chimeric antigen (Wright G J , Cherwinski H , Foster -Cuevas M , Brooke G , receptor comprising an extracellular domain specific for a Puklavec M J, Bigler M , Song Y , Jenmalm M , Gorman D , TRAIL receptor Tthe present invention relates to an iCAR McClanahan T , Liu M R , Brown M H , Sedgwick J D , ( or N CAR ) comprising an intracellular signaling domain Phillips J H , Barclay A N . 2003 " Characterization of the derived from a TRAIL receptor, and in particular from a CD200 receptor family in mice and humans and their TRAIL receptor selected from TRIOD ( other names: interactions with CD200 . J Immunol. > > 171 (6 ): 3034 -46 ) . TNFRSF10D :DCR2 , TRAILR4 , TRUNDD ) , TR10A ( TNF [0166 ] As preferred one, said polypeptide sequences of rereceptor superfamily member 10 , TRAIL -R1 or CD261) , or receptor have more than 80 % , preferably 90 % and more TR10B (TNF receptor superfamily member 10B , TRAIL preferably 95 % identity with SEQ ID NO : 36 (human cell R2 or CD262 ). surface glycoprotein CD200 receptor 1 ) . 0160 ] The present invention relates to an iCAR ( orN f 0167 ] The above sequences are listed in the following CAR ) comprising an intracellular signaling domain derived table 2 . US 2017 /0296623 A1 Oct . 19, 2017 17 TABLE 2 Amino - acid sequences of intracellular domain from a Tumor - necrosis - factor related apoptosis inducing ligand ( TRAIL ) receptor and cell surface glycoprotein CD200 receptor 1 UniProt SEQ Name entry ID NO : Amino - acid sequence TRIOD HUMAN 09UBN6 33 MGLWGQSVPTASSARAGRYPGARTASGTRPWLLDPKILKFVVFIVAVLLPVRVDSATIPRODEVPQ OTVAPOOORRSLKEEECPAGSHRSEYTGACNPCTEGVDYTIASNNLPSCLLCTVCKSGOTNKSSCT TTRDTVCOCEKGSFODKNSPEMCRTCRTGCPRGMVKVSNCTPRSDIKCKNESAASSTGKTPAAEET VTTILGMLASPYHYLIIIVVLVIILAVVVVGFSCRKKFISYLKGICSGGGGGPERVHRVLFRRRSC PSRVPGAEDNARNETLSNRYLOPTOVSEQEIOGOELAELTGVTVESPEEPORLLEOAEAE GCORRR ASATLEEGHAKETIQDQLVGSEKLFYEEDEAGSATSCLLLVPVNDADSADISTLLD TR10A HUMAN 000220 MAPPPARVHLGAFLAVTPNPGSAASGTEAAAATPSKVWGSSAGRIEPRGGGR GALPTSMGOHGPSA RARAGRAPGPRPAREASPRLRVHKTFKFVVVGVLLQVVPSSAATIKLHDOSIGTQQWEHSPLGELC PPGSHRSEHPGACNRCTEGVGYTNASNNLFACLPCTACKSDEEERSPCTTTRNTACOCKPGTFRND NSAEMCRKCSRGCPRGMVKVKDCTPWSDIECVHKESGNGHNIWVILVVTLVPLLLVAV LIVCCCI GSGCGGDPKCMDRVCFWRLGLLRGPGAEDNAHNEILSNADSLSTFVSEQQMESQEPADLTGVTVOS PGEAQCLLGPAEAEGSQRRRLLVPANGADPTETLMLFFDKFANIVPFDSWDQLMRQLDLT KNEIDV VRAGTAGPGDALYAMLMKWVNKTGRNASIHTLLDALERMEERHAREKIODLLVDSGKFI YLEDGTG SAVSLE TR1OB HUMAN 014763 35 MEQRGQNAPAASGARKRHGPGPREARGARPGPRVPKTLVLVVAAVLLLVSAESALITQQD LAPOOR AAPQOKRSSPSEGLCPPGHHISEDGRDCISCKYGODYSTHWNDLLFCLRCTRCDSGEVELSPCTTT RNTVCOCEEGTFREEDSPEMCRKCRTGCPRGMVKVGDCTPWSDIECVHKESGTKHSGEV PAVEETV TSSPGTPASPCSLSGIIIGVTVAAVVLIVAVFVCKSLLWKKVLPYLKGICSGGGGDPER VDRSSOR PGAEDNVLNEIVSILQPTQVPEQEMEVQEPAEPTGVNMLSPGESEHLLEPAEAERSQRRRLLVPAN EGDPTETLROCFDDFADLVPFDSWEPLMRKLGLMDNEIKVAKAEAAGHRDTLYTMLIKWVNKTGRD ASVHTLLDALETLGERLAKQKIEDHLLSSGKFMYLEGNADSAMS Cell surface Q8TD46 36 MLCPWRTANLGLLLILTIFLVAASSSLCMDEKQITQNYSKVLAEVNTSWPVKMATNAVLC CPPIAL glycoprotein RNLIIITWEIILRGQPSCTKAYRKETNETKETNCTDERITWVSRPDONSDLQIRPVAI THDGYYRC CD200 receptor IMVTPDGNFHRGYHLQVLVTPEVTLFONRNRTAVCKAVAGKPAAQISWIPEGDCATKQEYWSNGTV 1 human TVKSTCHWEVHNVSIVTCHVSHLTGNKSLYIELLPVPGAKKSAKLYIPYIILTIIILT IVGFIWLL KVNGCRKYKLNKTESTPVVEEDEMOPYASYTEKNNPLYDTTNKVKASEALOSEVDTDLHTL [0168 ] Extracellular Binding Domain or the N CAR that the one belonging to the P - CAR recognizes on - target According to the Invention cells ( i. e . tumoral cells ) and the one belonging to the N -CAR [0169 ] The inhibitory chimeric antigen receptor ( iCAR or recognizes off - target cells (healthy cells ) . Thus, when the N -CAR ) and the positive chimeric antigen receptor ( P - CAR ) engineered immune cell encounters a cancerous cell, only according to the present invention comprise an extracellular the P - CAR is able to bind to it and not the N -CAR , and ligand -binding domain . consequently the P -CAR can be activated and the cancerous [0170 ] The term “ extracellular ligand -binding domain ” as cell killed . In the other issue , when the engineered immune used herein is defined as an oligo - or polypeptide that is cell encounters a normal cell , both P -CAR and N -CAR can capable of binding a ligand . Preferably , the domain will be bind to it, and consequently the N -CAR can inactivate the capable of interacting with a cell surface molecule . For P -CAR : the normal cell will be preserved . example , the extracellular ligand -binding domain may be [0173 ] The antigen binding domain of the N - CAR can be chosen to recognize a ligand that acts as a cell surface any domain that binds to the off - tissue antigen including but marker on target cells associated with a particular disease not limited to a monoclonal antibody , a recombinant anti state . The combination of at least the two input signals body, a human antibody, a humanized antibody, and a corresponding to the recognition of different ligands by each functional fragment thereof. extracellular domains of said N -CAR and P -CAR allows the [0174 ] A humanized antibody can be produced using a inhibition of the P -CAR via the inhibitory transduction variety of techniques known in the art , including but not domain contained in the N -CAR . limited to , CDR - grafting ( see , e . g . , European Patent No . EP [0171 ] The system of the invention aims to avoid the “ off 239 , 400 ; International Publication No. WO 91/ 09967 ; and target” events , wherein the engineered immune cells target U . S . Pat . Nos. 5 , 225 ,539 , 5 ,530 , 101, and 5 , 585 ,089 , each of not only tumoral cells due in particularly to lack of speci which is incorporated herein in its entirety by reference ) , ficity of the antigen ( the latter being present on the cancerous veneering or resurfacing ( see , e . g ., European Patent Nos . EP cells but can also be present on normal cells ) . 592 , 106 and EP 519 ,596 ; Padlan , 1991 , Molecular Immu [0172 ] Therefore , the extracellular binding domains nology, 28 (4 /5 ): 489 -498 ; Studnicka et al. , 1994 , Protein within the scope of the invention are chosen in such a way Engineering, 7 (6 ): 805 -814 ; and Roguska et al. , 1994 , US 2017 /0296623 A1 Oct . 19, 2017 PNAS , 91 :969 - 973 , each of which is incorporated herein by Natl. Acad . Sci . USA 85 :5879 -5883 ). ScFv molecules can be its entirety by reference ), chain shuffling (see , e . g ., U . S . Pat. produced by linking VH and VL regions together using No . 5 , 565 ,332 , which is incorporated herein in its entirety by flexible polypeptide linkers . The scFv molecules comprise a reference ) , and techniques disclosed in , e . g ., U . S . Patent linker ( e . g . , a SerGly linker ) with an optimized length and / or Application Publication No . US2005 /0042664 , U . S . Patent amino acid composition . The linker length can greatly affect Application Publication No . US2005 /0048617 , U . S . Pat. how the variable regions of a scFv fold and interact . In fact , No . 6 ,407 ,213 , U .S . Pat . No . 5 ,766 ,886 , International Pub if a short polypeptide linker is employed ( e . g ., between 5 - 10 lication No. WO 9317105 , Tan et al . , J . Immunol. , 169: amino acids) intrachain folding is prevented . Interchain 1119 - 25 (2002 ) , Caldas et al . , Protein Eng ., 13 ( 5 ) :353 -60 folding is also required to bring the two variable regions ( 2000 ) , Morea et al . , Methods, 20 ( 3 ) : 267 -79 ( 2000 ), Baca et together to form a functional epitope binding site . For al. , J . Biol. Chem . , 272 ( 16 ) : 10678 - 84 ( 1997 ) , Roguska et examples of linker orientation and size see , e . g ., Hollinger et al. , Protein Eng ., 9 ( 10 ) : 895 - 904 ( 1996 ) , Couto et al. , Cancer al. 1993 Proc Natl Acad . Sci. U . S . A . 90 :6444 -6448 , U . S . Res. , 55 (23 Supp ) : 5973s - 5977s ( 1995 ) , Couto et al . , Cancer Patent Application Publication Nos. 2005 / 0100543 , 2005/ Res. , 55 (8 ): 1717 - 22 ( 1995 ) , Sandhu JS , Gene , 150 ( 2 ): 409 0175606 , 2007 /0014794 , and PCT publication Nos. 10 ( 1994 ) , and Pedersen et al. , J. Mol. Biol. , 235 ( 3 ) : 959 -73 WO2006 /020258 and WO2007 /024715 , is incorporated ( 1994 ) , each of which is incorporated herein in its entirety herein by reference . by reference . Often , framework residues in the framework [0179 ] An scFv can comprise a linker of at least 1, 2 , 3 , 4 , regions will be substituted with the corresponding residue 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 25 , from the CDR donor antibody to alter, for example improve , 30 , 35 , 40 , 45 , 50 , or more amino acid residues between its antigen binding . These framework substitutions are identi VL and VH regions . The linker sequence may comprise any fied by methods well -known in the art, e . g . , by modeling of naturally occurring amino acid . The linker sequence may the interactions of the CDR and framework residues to comprise amino acids glycine and serine. The linker identify framework residues important for antigen binding sequence may comprise sets of glycine and serine repeats and sequence comparison to identify unusual framework such as (Gly4Ser ) n , where n is a positive integer equal to or residues at particular positions. ( See , e .g . , Queen et al. , U . S . greater than 1 . The linker can be (Gly4Ser )4 or (Gly4Ser ) 3 . Pat. No . 5 ,585 , 089 ; and Riechmann et al ., 1988 , Nature , Variation in the linker length may retain or enhance activity , 332 : 323 , which are incorporated herein by reference in their giving rise to superior efficacy in activity studies . entireties. ) . [0180 ] In a preferred embodiment, the antigen binding [0175 ] In a preferred embodiment, said extracellular domain of the N -CAR comprises an scFv . The off - tissue ligand -binding domain is a single chain antibody fragment antigen recognized by the antigen binding domain of the ( scFv ) . The latter comprises usually the light (VD ) and the N -CAR is preferably an antigen that is not present or present heavy (Vy variable fragment of a target antigen specific at low level on the tumour cells targeted by the P -CAR . monoclonal antibody joined by a flexible linker . Other [0181 ] In a preferred embodiment, the antigen binding binding domain than scFv can also be used for predefined domain of the N - CAR comprises a scFv . The off- tissue targeting of lymphocytes , such as camelid single - domain antigen recognized by the antigen binding domain of the antibody fragments , receptor ligands like a vascular N -CAR is preferably an antigen that is not present or present endothelial growth factor polypeptide, an integrin - binding at low level on the tumour cells targeted by the P -CAR and peptide , heregulin or an IL - 13 mutein , antibody binding is expressed in normal tissue ( non precancerous or non domains , antibody hypervariable loops or CDRs as non cancerous ) . limiting examples . [0182 ] By “ cancerous or tumor cell” , it is meant cells [ 0176 ] In some embodiments , the antibody binding differing from normal cells in many ways that allow them to domain is a Fv , a Fab , a ( Fab ' ) 2 , or a bi- functional ( e. g . grow out of control and become invasive . Cancer cells are bi- specific ) hybrid antibody ( e . g ., Lanzavecchia et al. , Eur. less specialized than normal cells and continue to divide J. Immunol. 17 , 105 ( 1987 ) ) . In some embodiments , the without stopping . They are able to ignore signals that antigen binding domain of the N -CAR of the invention binds normally tell cells to stop dividing or that begin a process an off - tissue antigen with wild - type or enhanced affinity . known as programmed cell death , or apoptosis , which the [0177 ] By " affinity ” is meant a measure of binding body uses to get rid of unneeded cells . strength . Without being bound to theory , affinity depends on [0183 ] Present at low level on the tumour cells targeted by the closeness of stereochemical fit between antibody com the P -CAR means that the expression of said off - tissue bining sites and antigen determinants, on the size of the area antigen is undetectable in tumor cells using any known of contact between them , and on the distribution of charged technique of antigen detection ( eg flow cytometry , himmu and hydrophobic groups . Affinity also includes the term nohisto chemistry , western blot) or represents less than 10 % “ avidity , " which refers to the strength of the antigen -anti expression as compared to expression in a cell or a tissue body bond after formation of reversible complexes. Methods used as a positive control. for calculating the affinity of an antibody for an antigen are [0184 ] In one embodiment said the antigen binding known in the art , including use of binding experiments to domain of the N -CAR comprises at least a scFv specific for calculate affinity . Antibody activity in functional assays any one of the following antigen CD56 , CD205 , CD83 , ( e . g . , flow cytometry assay ) is also reflective of antibody CD206 , CD200 , CD36 , RARRES1, Troponin C , Beta - 1 affinity . Antibodies and affinities can be phenotypically integrin , CCKBR , GALR1, CD4 , CD20 , CD22 , CD25 , characterized and compared using functional assays (e . g , MUC1 antigen CD20 . flow cytometry assay ). [0185 ] In one embodiment said the antigen binding [0178 ] In some instances, scFvs can be prepared according domain of the N -CAR comprises at least a scFv specific for to method known in the art ( see , for example , Bird et al. , any one of the following antigen CD56 , CD205 , CD83 , ( 1988 ) Science 242 :423 -426 and Huston et al. , ( 1988 ) Proc . CD206 , CD200 , CD36 , or RARRES1 . US 2017 /0296623 A1 Oct . 19 , 2017 19 [0186 ] In one embodiment said the antigen binding [0190 ] In the present invention , the antigen binding domain of the N -CAR comprises at least a scFv specific for domain of the N -CAR binds to a cell -surface protein present any one of the following antigen Troponin C , Beta - 1 inte in normal tissue but not present or present at lower level on grin , CCKBR , or GALR1. a tumor as compared to a the same cell in normal tissue said [0187 ] In one embodiment said the antigen binding binding domain binds to an off -tissue antigen . domain of the N -CAR comprises a scFv specific for any one [0191 ] N -CAR antigens could also include antigens that of the following antigen CD4, CD20 , CD22 , CD25 , MUC1 are independent of the antigen that the P - CAR is targeting antigen . In one embodiment said the antigen binding domain and that are down- regulated in tumor of interest , but present of the N -CAR comprises at least a scFv specific for any one in all normal tissues of concern . Examples of such antigens of the following antigen CD20 , PSMA , BCMA , CD19The for pancreatic ductal adenocarcinoma are TMPRSS11B , below table 3 provides examples of combinations of N -CAR CYP17A1 and ATP4B and examples of such antigens for and P -CAR antigens . Combinations of a P -CAR directed to kidney clear cell carcinoma are GP2 , MUC21, CLCA4 and anti - D33 , FLT3 , MUC16 , and to anti- MUC17 CAR with SLC27A6 . their N -CARs counterparts , are not part of the present [0192 ] In certain embodiments , the subject has metastatic invention . breast cancer, hematological malignancy, or a solid tumor, P - CAR Antigen N -CAR Antigen CD38 CD56 antigen : expression on the surface of neurons , glia , skeletal muscle and natural killer cells CD205 antigen : expression on cortical thymic epithelial cells and by dendritic cell (DC ) subsets CD83 antigen : expression on activated lymphocytes , Langerhans cells and interdigitating reticulum cells CD206 antigen : expression on the surface of macrophages and dendritic cells , on the surface of skin cells such as human dermal fibroblasts and keratinocytes CD200 antigen : expression on cells originating from the hematopoietic cells , activated T cells , endothelial neuronal cells and cells of the reproductive organs (ovaries and placental trophoblasts ) CD36 antigen : expression in adipocytes endothelial cells and monocytes RARRES1 antigen : expression of this gene upregulated by tazarotene as well as by retinoic acid receptors csi Troponin C antigen : expression in heart Beta - 1 integrin antigen : expression in endothelial cells and fibroblasts ( at protein level ). Expression in intestine , colon , testis , ovary , thymus, spleen and prostate CCKBR antigen : expression in stomach , pancreas, brain and gallbladder GALR1 antigen : expression in adrenal gland CUBN antigen : expression in kidney and small intestine CD123 CD4 antigen : expression in appendix , bone marrow , lymph node, tonsil and spleen CD20 antigen : expression mainly in spleen appendix and lymph node CD22 antigen : expression in particular in appendix , lymph node , tonsil and spleen CD25 antigen : expression mainly in bladder and lymph node MUC1 antigen : expression in kidney ROR1 Troponin C antigen : expression in heart Beta - 1 integrin antigen : expression in endothelial cells and fibroblasts (at protein level) . Expression in intestine, colon , testis , ovary , thymus, spleen and prostate CCKBR antigen : expression in stomach , pancreas , brain and gallbladder GALR1 antigen : expression in adrenal gland MUC1 antigen : expression in kidney CD33 Antigens specifically expressed in dendritic cells and /or haematopoetic stem cells such as ITGAX , CD1E , CD34 , CD1C , CD123 , CD141 FLT3 Antigens specifically expressed in haematopoetic stem cells such as CD34 or specifically expressed in Brain cerebellum such as ZP2 , GABRA , CRTAM , GRM4, MDGA1 MSLN Antigens specifically expressed in lung such as SFTPC , ROS1 , SLC6A4, AGTR2 MUC16 Antigens specifically expressed in salivary gland such as LRRC26 , HTR3A , TMEM211, MRGPRX3 MUC17 Antigens specifically expressed in colon & small intestine such as MEP1B , TMIGD1 , CEACAM20 , ALPI CD20 CD20 [0188 ] Extracellular- Binding Domain of N -CAR Accord and the human leukocyte antigen (HLA ) is HLA - I . In certain ing to the Invention embodiments , the subject has a tumor that has undergone [ 0189 ] In the various embodiments of the aspects delin epithelium to mesenchymal transition ( EMT) , and the anti eated herein , the binding of an antigen to the NCAR gen is one or more of an Epithelial- mesenchymal transition activates the intracellular signaling domain resulting in a (EMT ) antigen , E - cadherin , and cytokeratin . In various decrease in an immune response, preferably in the CTL embodiments , the binding of the inhibitory chimeric antigen activity . receptor and the antigen , decreases cell death in a cell US 2017 /0296623 A1 Oct . 19, 2017 20 comprising the antigen . The method can reduce graft versus 1917[0199 ] The N - CAR of the present invention is a trans host disease (GVHD ) in the subject , or a symptom thereof. membrane polypeptide containing at least: [0193 ] Extracellular -Binding Domain of P -CAR [0200 ] an extracellular binding domain ; [0194 ] The extracellular ligand - binding domain of a P [ 0201] a transmembrane domain and , CAR according to the present invention can also comprise a [ 0202 ] an intracellular domain comprising at least one peptide binding an antigen of the target, a peptide or a polypeptide sequence involved in transduction signal, protein binding an antibody that binds an antigen of the preferably an inhibitory transduction signal said poly target, a peptide or a protein ligand such as a growth factor, peptide sequence is not significantly expressed in a cytokine or a hormone as non -limiting examples binding T -cell , and / or said polypeptide sequence is from a a receptor on the target , or a domain derived from a receptor ( TRAIL ) receptor and / or from a CD200 receptor 1 , such as a growth factor receptor, a cytokine receptor or a provided that said polypeptide sequence is not a hormone receptor as non -limiting examples , binding a pep sequence selected from group consisting of SEQ ID tide or a protein ligand on the target. Preferably the target is NO : 13 (human SIGL8 ) , SEQ ID NO : 14 (human a cell. SIGL7 ), SEQ ID NO : 17 (human SIGL5) , SEQ ID [0195 ] As non - limiting example , the ligand of the target NO : 20 (human SIGLI ) , SEQ ID NO : 21 ( human can be a tumor- associated surface antigen , such as ErbB2 SIGL6 ), SEQ ID NO : 22 (human CD33 ), SEQ ID (HER2 /neu ) , carcinoembryonic antigen (CEA ), epithelial NO : 26 (human SIG12 ), SEQ ID NO :31 (human cell adhesion molecule ( EpCAM ) , epidermal growth factor SIG11 ) , SEQ ID NO : 32 ( human SIG10 ) and SEQ ID receptor (EGFR ) , CD19 , CD20 , CD30 , CD40 , disialogan NO : 19 (human PECA1) . glioside GD2 , GD3, C - type lectin - likemolecule - 1 ( CLL - 1 ) , [0203 ] In a preferred embodiment, said intracellular ductal -epithelial mucine , gp36 , TAG - 72 , glycosphingolip domain comprises a polypeptide sequence also called inhibi ids , glioma - associated antigen , B -human chorionic gonado tory transduction domain . By " inhibitory transduction tropin , alphafetoprotein ( AFP ) , lectin - reactive AFP , thyro domain ” , it is meant here a transmembrane polypeptide globulin , RAGE - 1, MN -CA IX , human telomerase reverse which contains a region encoding for an inhibitory trans transcriptase , RUI, RU2 (AS ) , intestinal carboxyl esterase , duction signal . mut hsp70 - 2 , M -CSF , prostase , prostase specific antigen [ 0204 ] In a preferred embodiment said inhibitory trans (PSA ), PAP, NY - ESO - 1, LAGA - la , p53 , prostein , PSMA, duction signal attenuates the activity of the immune cells , in surviving and telomerase , prostate - carcinoma tumor anti particular of the CTL activity , preferably a CTL activity gen - 1 (PCTA - 1 ), MAGE , ELF2M , neutrophil elastase , eph induced upon binding of a P - CAR of the invention . rin B2 , CD22 , insulin growth factor ( IGF1 ) - I, IGF - II , IGFI [ 0205 ] According to a preferred embodiment, the N -CAR receptor, mesothelin , a major histocompatibility complex comprises at least: (MHC ) molecule presenting a tumor- specific peptide epitope , 5T4 , ROR1, Nkp30 , NKG2D , tumor stromal anti [ 0206 ] an extracellular binding domain ; gens, the extra domain A (EDA ) and extra domain B (EDB ) [0207 ] a transmembrane domain and , of fibronectin and the A1 domain of tenascin - C ( TnC A1) [ 0208 ] an intracellular domain comprising at least one and fibroblast associated protein ( fap ) , LRP6 , melamona polypeptide sequence involved in transduction signal, associated Chondroitin Sulfate Proteoglycan (MCSP ) , preferably an inhibitory transduction signal said poly CD38 / CS1, MART1, WT1, MUC1, LMP2 , Idiotype, NY peptide sequence is not significantly expressed in ESO - 1 , Ras mutant, gp100 , proteinase 3 , bcr -abl , tyrosinase , T -cell , and /or said polypeptide sequence is from a hTERT, EphA2, ML - TAP, ERG , NA17 , PAX3 , ALK , ( TRAIL ) receptor and / or from a CD200 receptor 1 , Androgen receptor; a lineage - specific or tissue specific provided that said polypeptide sequence is not a antigen such as CD3, CD4 , CD8, CD24 , CD25 , CD34 , sequence selected from group consisting of SEQ ID CD79 , CD116 , CD117 , CD135 , CD123 , CD133 , CD138 , NO : 13 (human SIGL8) , SEQ ID NO : 14 (human CTLA -4 , B7 - 1 (CD80 ), B7 - 2 ( CD86 ), endoglin , a major SIGL7 ) , SEQ ID NO : 17 (human SIGL5 ) , SEQ ID histocompatibility complex (MHC ) molecule , BCMA NO : 20 (human SIGLI) , SEQ ID NO : 21 (human (CD269 , TNFRSF 17 ) , or a virus- specific surface antigen SIGL6 ), SEQ ID NO :22 (human CD33 ), SEQ ID such as an HIV - specific antigen ( such as HIV gp120 ) ; an NO : 26 (human SIG12 ) , SEQ ID NO :31 (human EBV - specific antigen , a CMV- specific antigen , a HPV - spe SIG11 ), SEQ ID NO : 32 (human SIG10 ) and SEQ ID cific antigen , a Lasse Virus -specific antigen , an Influenza NO : 19 ( human PECA1) Virus -specific antigen as well as any derivate or variant of [ 0209 ] wherein said inhibitory transmembrane polypep these surface markers . In specific cases , the ligand that the tide comprises a sequence with more than 80 % , pref chimeric antigen receptor recognizes is present on the sur erably 90 % and more preferably 95 % identity with face of a target cell , particularly cancer cell or viral cell. In SEQ ID NO : 1 (human KI2L2 ), SEQ ID NO : 2 (human some embodiments , the ligand that the chimeric antigen KI2L1 ) , SEQ ID NO :3 (human FCG2B ), SEQ ID NO : 4 receptor recognizes is present in a tumor microenvironment. (human KI2L3 ), SEQ ID NO : 5 (human KI3L2 ), SEQ In some aspects of the invention , the ligand that the chimeric ID NO : 6 (human KI2L4) , SEQ ID NO : 7 (human antigen receptor recognizes is a growth factor. KI3L1) , SEO ID NO : 8 ( human KIZLA ) , SEO ID NO : 9 (human MILR1) , SEQ ID NO : 10 (human LIRB4) , [0196 ] In a preferred embodiment, CD33 , BCMA and SEO ID NO : 11 ( human LIRB3 ) , SEO ID NO : 12 ( hu EGFRVIII do not belong to the present invention . man KI3L3 ) , SEQ ID NO : 15 (human LIRB5) , SEQ ID [0197 ] N - CAR Architecture & its Other Components NO : 16 (human LIRB2 ) , SEQ ID NO : 18 (human [0198 ] The N - CAR of the invention may have the single FCRL4 ) , SEQ ID NO :23 (human FCRL5 ) , SEQ ID chain or the multi- chain architecture . The multi- chain con NO : 24 (human FCRL2) , SEQ ID NO : 25 (human formation is disclosed in WO2014039523 . FCRL1) , SEQ ID NO : 27 (human FCRL3 ), SEQ ID US 2017 /0296623 A1 Oct . 19 , 2017 NO : 28 (human MPZL1) , SEQ ID NO : 29 ( human [0237 ] According to one more preferred embodiment, the PILRA ) or SEQ ID NO :30 (human PVR ) . N -CAR comprises at least: [ 0210 ] According to a more preferred embodiment, the [ 0238 ] an extracellular binding domain ; N - CAR comprises at least: [0239 ] a transmembrane domain and , [0211 ] an extracellular binding domain ; 10240 ] an intracellular domain comprising a sequence [ 0212 ] a transmembrane domain and , of SEQ ID NO : 33 (human TR10D ) . 02131 an intracellular domain comprising [0241 ] According to one more preferred embodiment, the [ 0214 ] an inhibitory transmembrane polypeptide com N -CAR comprises at least: prising a sequence selected from the list consisting of [ 0242 ] an extracellular binding domain ; SEQ ID NO : 1 (human KI2L2 ) , SEQ ID NO : 2 (human 10243 ] a transmembrane domain and , KI2L1) , SEQ ID NO : 3 (human FCG2B ) , SEQ ID NO : 4 10244 ] an intracellular domain comprising a sequence (human KI2L3 ) , SEQ ID NO : 5 (human KI3L2 ) , SEQ with more than 80 % , preferably 90 % and more pref ID NO : 6 (human KI2L4 ), SEQ ID NO : 7 (human erably 95 % identity with SEQ ID NO : 34 (human KI3L1) , SEQ ID NO : 8 (human KIZLA ), SEQ ID NO : 9 TR10A ). (human MILR1) , SEQ ID NO : 10 (human LIRB4) , [0245 ] According to one more preferred embodiment, the SEQ ID NO : 11 (human LIRB3 ) , SEQ ID NO : 12 (hu N -CAR comprises at least: man KI3L3 ) , SEQ ID NO : 15 (human LIRB5 ) , SEQ ID (0246 ] an extracellular binding domain ; NO : 16 (human LIRB2) , SEQ ID NO : 18 (human 102471 a transmembrane domain and , FCRL4 ) , SEQ ID NO : 23 (human FCRL5 ) , SEQ ID [0248 ] an intracellular domain comprising a sequence NO : 24 (human FCRL2 ) , SEQ ID NO : 25 (human of SEQ ID NO : 34 (human TR10A ) . FCRL1) , SEQ ID NO : 27 (human FCRL3) , SEQ ID [0249 ] According to one more preferred embodiment , the NO : 28 (human MPZL1) , SEQ ID NO :29 (human N -CAR comprises at least: PILRA ) or SEQ ID NO :30 (human PVR ) . [0250 ] an extracellular binding domain ; [0215 ] According to an embodiment, the N -CAR com [ 0251] a transmembrane domain and , prises at least : an extracellular binding domain ; [0252 ] an intracellular domain comprising a sequence [0216 ] a transmembrane domain and , with more than 80 % , preferably 90 % and more pref [ 0217 ] an intracellular domain comprising erably 95 % identity with SEQ ID NO : 35 (human [ 0218 ] an inhibitory transmembrane polypeptide com TR10B ). prising a sequence with more than 80 % , preferably [0253 ] According to one more preferred embodiment, the 90 % and more preferably 95 % identity with SEQ ID N -CAR comprises at least: [ 0254 ] an extracellular binding domain ; NO :36 (CD200 receptor 1 ) . [0255 ] a transmembrane domain and , [ 0219] According to a more preferred embodiment, the [0256 ] an intracellular domain comprising a sequence N -CAR comprises at least: of SEQ ID NO : 35 (human TR10B ). [ 0220 ] an extracellular binding domain ; [0257 ] According to one preferred embodiment, the [0221 ] a transmembrane domain and , N -CAR of the present comprising : [0222 ] an intracellular domain comprising [0258 ] an extracellular domain comprising an antigen [0223 ] a sequence of SEQ ID NO :36 (CD200 receptor binding domain ; 1 ) . [ 0259 ) a transmembrane domain ; [0224 ] According to a preferred embodiment, the N -CAR [0260 ] an intracellular domain ; comprises at least: [0261 ] wherein said intracellular domain comprises a [0225 ] an extracellular binding domain ; polypeptide sequence consisting essentially of aminoacids [0226 ] a transmembrane domain and, No 201 - 375 from SEQ ID NO : 8 (human KIZLA ) . [ 0227 ] an intracellular domain comprising a sequence [0262 ] By “ polypeptide sequence consisting essentially with more than 80 % , preferably 90 % and more pref of” , it is meant that the polypeptide is the one identical to the erably 95 % identity with SEQ ID NO : 33 (human part of the inhibitory molecule which is used in the N -CARs TRIOD ) , SEO ID NO : 34 ( human TR10A ) or SEO ID presented here . However, at least one to a few amino acid NO : 35 (human TR10B ) . substitution (s ) is (are ) contemplated within the present [0228 ] According to an even more preferred embodiment, invention in order to bring a modulation of its inhibitory the N -CAR comprises at least : function in case of need . [0229 ] an extracellular binding domain ; [0263 ] According to one preferred embodiment, the [0230 ] a transmembrane domain and , N -CAR of the present comprising : [0231 ] an intracellular domain comprising a sequence [0264 ] an extracellular domain comprising an antigen selected from the list consisting of binding domain ; [0232 ] SEQ ID NO : 33 (human TR10D ), SEQ ID NO : 34 0265 ] a transmembrane domain ; (human TR10A ) and SEQ ID NO : 35 (human TR10B ) . [0266 ] an intracellular domain ; [ 0233] According to one more preferred embodiment , the [0267 ] wherein said intracellular domain comprises a N - CAR comprises at least: polypeptide sequence consisting essentially of amino [0234 ] an extracellular binding domain ; acids No 206 - 348 from SEQ ID NO : 2 (human [0235 ] a transmembrane domain and , KIR2DL1) . [0236 ] an intracellular domain comprising a sequence [0268 ] According to one preferred embodiment, the with more than 80 % , preferably 90 % and more pref N -CAR of the present comprising : erably 95 % identity with SEQ ID NO : 33 (human [ 0269 ] an extracellular domain comprising an antigen TRIOD ) . binding domain ; US 2017 /0296623 A1 Oct . 19 , 2017 [0270 ] a transmembrane domain ; [ 0303 ] According to one preferred embodiment, the [0271 ] an intracellular domain ; N -CAR of the present comprising : [0272 ] wherein said intracellular domain comprises a [0304 ] an extracellular domain comprising an antigen polypeptide sequence consisting essentially of amino binding domain ; acids No 206 - 348 from SEQ ID NO : 1 (human [0305 ] a transmembrane domain ; KIR2DL2 ) . 10306 ] an intracellular domain ; [ 0273] According to one preferred embodiment, the [0307 ] wherein said intracellular domain comprises a N -CAR of the present comprising : polypeptide sequence consisting essentially of amino [0274 ] an extracellular domain comprising an antigen acids No 214 -343 from SEQ ID NO : 9 (human MILR1) . binding domain ; [0308 ] According to one preferred embodiment, the [0275 ] a transmembrane domain ; N -CAR of the present comprising : 0276 ] an intracellular domain ; [ 0309 ] an extracellular domain comprising an antigen 0277 ] wherein said intracellular domain comprises a binding domain ; polypeptide sequence consisting essentially of amino [0310 ) a transmembrane domain ; 10311 an intracellular domain ; acids No 206 - 341 from SEQ ID NO : 4 (human [ 0312 ] wherein said polypeptide sequence of the recep KIR2DL3 ). tor of amino acids No 216 - 448 from SEQ ID NO : 10 [0278 ] According to one preferred embodiment, the (human LIRB4) . N -CAR of the present comprising : [0313 ] According to one preferred embodiment, the [0279 ] an extracellular domain comprising an antigen N -CAR of the present comprising: binding domain ; [0314 ] an extracellular domain comprising an antigen [0280 ] a transmembrane domain ; binding domain ; [0281 ] an intracellular domain ; (0315 ] a transmembrane domain ; [0282 ] wherein said intracellular domain comprises a [0316 ] an intracellular domain ; polypeptide sequence consisting essentially of amino [03171 wherein said intracellular domain comprises a acids No 206 - 348 from SEQ ID NO : 2 (human polypeptide sequence consisting essentially of amino KIR2DL1) . acids No 420 -631 from SEQ ID NO : 11 (human [ 0283 ] According to one preferred embodiment, the LIRB3) . N - CAR of the present comprising: [0318 ] According to one preferred embodiment, the 10284 ] an extracellular domain comprising an antigen N -CAR of the present comprising : binding domain ; [0319 ] an extracellular domain comprising an antigen [0285 ] a transmembrane domain ; binding domain ; [0286 ] an intracellular domain ; [ 0320 ] a transmembrane domain ; [0287 ] wherein said intracellular domain comprises a [0321 ] an intracellular domain ; polypeptide sequence consisting essentially of amino [0322 ] wherein said intracellular domain comprises a acids No 203 -377 from SEQ ID NO :6 (human polypeptide sequence consisting essentially of amino acids KIR2DL4 ) . No 296 -410 from SEQ ID NO : 12 (human KI3L3) . [ 0288 ] According to one preferred embodiment, the [0323 ]. According to one preferred embodiment, the N -CAR from the present comprising : N -CAR of the present comprising: [ 0289 ] an extracellular domain comprising an antigen [ 0324 ] an extracellular domain comprising an antigen binding domain ; binding domain ; [0290 ] a transmembrane domain ; [0325 ] a transmembrane domain ; [0291 ] an intracellular domain ; 0326 ] an intracellular domain ; [ 02921 wherein said intracellular domain comprises a [0327 ] wherein said intracellular domain comprises a polypeptide sequence consisting essentially of amino acids polypeptide sequence consisting essentially of amino acids No 301 -444 from SEQ ID NO : 7 ( human KIR3DL1) . No 419 - 590 from SEQ ID NO : 15 (human LIRB5 ) . 0293] According to one preferred embodiment, the [0328 ] According to one preferred embodiment, the N -CAR of the present comprising : N -CAR of the present comprising: [ 0294 ] an extracellular domain comprising an antigen [0329 ] an extracellular domain comprising an antigen binding domain ; binding domain ; [0295 ] a transmembrane domain ; [0330 ] a transmembrane domain ; [0296 ] an intracellular domain ; [ 0331 ] an intracellular domain ; [0297 ] wherein said intracellular domain comprises a [0332 ] wherein said intracellular domain comprises a polypeptide sequence consisting essentially of amino acids polypeptide sequence consisting essentially of amino acids No 301 - 455 from SEQ ID NO : 5 ( human KIR3DL2 ) . No 420 -598 from SEQ ID NO : 16 (human LIRB2 ). [0298 ] According to one preferred embodiment, the [ 0333 ] According to one preferred embodiment, the N -CAR of the present comprising : N -CAR of the present comprising: [0299 ] an extracellular domain comprising an antigen [0334 ] an extracellular domain comprising an antigen binding domain ; binding domain ; [0300 ] a transmembrane domain ; [0335 ] a transmembrane domain ; [0301 ] an intracellular domain ; 10336 an intracellular domain ; [0302 ] wherein said intracellular domain comprises a [0337 ] wherein said intracellular domain comprises a polypeptide sequence consisting essentially of amino acids polypeptide sequence consisting essentially of amino acids No 204 - 310 from SEQ ID NO :24 (human FRGR2B ) . No 375 -515 from SEQ ID NO : 18 ( human FCRL4 ). US 2017 /0296623 A1 Oct . 19 , 2017 22 [0338 ] According to one preferred embodiment, the [0373 ] According to one preferred embodiment, the N -CAR of the present comprising : N -CAR of the present comprising : [0339 ] an extracellular domain comprising an antigen [0374 ] an extracellular domain comprising an antigen binding domain ; binding domain ; [0340 ) a transmembrane domain ; [0375 ] a transmembrane domain ; [0341 ] an intracellular domain ; 10376 ] an intracellular domain ; [0342 ] wherein said intracellular domain comprises a f0377 ] wherein said intracellular domain comprises a polypeptide sequence consisting essentially of amino acids polypeptide sequence consisting essentially of amino acids No 753 - 977 from SEQ ID NO :23 (human FCRL5 ) . No 229 - 325 from SEQ ID NO : 36 ( human CD200 recep [ 0343] According to one preferred embodiment, the tor1torl ) .. N -CAR of the present comprising : [0378 ] According to a preferred embodiment, the N - CAR [0344 ] an extracellular domain comprising an antigen of the present comprising : binding domain ; [ 0379 ] an extracellular domain comprising an antigen [0345 ) a transmembrane domain ; binding domain ; [0346 ] an intracellular domain ; [0380 ] a transmembrane domain ; [0347 ] wherein said intracellular domain comprises a [ 0381 ] an intracellular domain ; polypeptide sequence consisting essentially of amino acids [0382 ] wherein said intracellular domain comprises a No 388 - 508 from SEQ ID NO :24 (human FCRL2) . polypeptide sequence consisting essentially of amino acids [0348 ] According to one preferred embodiment, the N -CAR of the present comprising: No 181 - 386 from SEQ ID NO : 33 (human TR10D ). [ 0349 ] an extracellular domain comprising an antigen [ 0383 ] According to a preferred embodiment, the N -CAR binding domain ; of the present comprising: 10350 a transmembrane domain ; [0384 ] an extracellular domain comprising an antigen [0351 ] an intracellular domain ; binding domain ; [0352 ] wherein said intracellular domain comprises a [0385 ] a transmembrane domain ; polypeptide sequence consisting essentially of amino acids 10386 ] an intracellular domain ; No 292 - 429 from SEQ ID NO : 25 (human FCRL1 ) . [0387 ] wherein said intracellular domain comprises a [ 0353] According to one preferred embodiment, the polypeptide sequence consisting essentially of amino acids N - CAR of the present comprising : No 230 - 468 from SEQ ID NO : 34 (human TR10A ) [0354 ] an extracellular domain comprising an antigen [0388 ] According to a preferred embodiment, the N - CAR binding domain ; of the present comprising : [0355 ) a transmembrane domain ; [0389 ] an extracellular domain comprising an antigen [0356 ] an intracellular domain ; binding domain ; [ 0357 ] wherein said intracellular domain comprises a [ 0390 ] a transmembrane domain ; polypeptide sequence consisting essentially of amino acids 0391 ] an intracellular domain ; No 564 -734 from SEQ ID NO : 27 (human FCRL3 ). [0392 ] wherein said intracellular domain comprises a [0358 ] According to one preferred embodiment, the polypeptide sequence consisting essentially of amino acids N -CAR of the present comprising: No 179 - 440 from SEQ ID NO : 35 ( human TR10B ). [0359 ] an extracellular domain comprising an antigen [0393 ] According to another preferred embodiment, the binding domain ; N -CAR comprises at least : 10360 ) a transmembrane domain , [ 0394 ) an extracellular binding domain ; [0361 ] an intracellular domain ; [0395 ] an transmembrane domain , and ; [ 0362 ] wherein said intracellular domain comprises a 03961 a linker between the extracellular binding polypeptide sequence consisting essentially of amino acids domain and the transmembrane domain , No 147 -269 from SEQ ID NO : 28 (human MPZL1) . [0397 ] said linker can be any one known by the skilled [ 0363] According to one preferred embodiment, the man in the art. N - CAR of the present comprising : 10398 ] Preferably, this linker is a GS linker 1 or a GS [0364 ] an extracellular domain comprising an antigen linker 2 comprising a sequences of SEQ ID NO :39 and SEQ binding domain ; ID NO : 40 , more preferably this linker is a GS linker 1 or a [ 0365 ] a transmembrane domain ; GS linker 2 consisting in a sequences of SEQ ID NO : 39 and [0366 ] an intracellular domain ; SEQ ID NO :40 . 10367 ] wherein said intracellular domain comprises a [0399 ] Therefore , the extracellular part of the N -CAR may polypeptide sequence consisting essentially of amino comprises: acids No 151- 303 from SEQ ID NO :29 (human [0400 ] an extracellular- binding domain comprising at PILRA ) . least one scFvs from a monoclonal antibody for binding [ 0368 ] According to one preferred embodiment, the to “ off - target ” antigen expressed on healthy cells ; and N - CAR of the present comprising : preferably said off - target” antigen is not expressed on [0369 ] an extracellular domain comprising an antigen cells targeted by the P -CAR . a transmembrane domain binding domain ; and ; [0370 ] a transmembrane domain ; [ 0401 ] a linker binding together the two previous com [0371 ] an intracellular domain ; ponents . [0372 ] wherein said intracellular domain comprises a [0402 ] Said above scFvs of a monoclonal antibody binds polypeptide sequence consisting essentially of amino acids preferably to " off - target ” antigens expressed in healthy No 329 - 417 from SEQ ID NO :30 (human PVR ) . tissues or healthy cells . US 2017 /0296623 A1 Oct . 19, 2017 24 [0403 ] For instance , for the treatment of acute myeloid [ 0417 ] In another embodiment, the N -CAR of the inven leukemia ( AML ) , when the antigen targeted by the P -CAR tion comprises at least : is CD123 , said extracellular- binding domain of the N -CAR [0418 ] an extracellular domain comprising at least one binds to at least one “ off - target ” antigen expressed on scFv from an antibody binding specifically to an anti healthy cells or healthy immune cells that may be chosen gen selected from CD4 antigen , CD20 antigen , CD22 amongst an antigen selected from CD4 antigen ( expressed in antigen , CD25 antigen and /or MUC1 antigen ; appendix , bone marrow , lymph node, tonsil and spleen ) , [0419 ] a linker with a SEQ ID NO :38 , SEQ ID NO :39 CD20 antigen ( expressed mainly in spleen appendix and or SEQ ID NO : 40 ; and , lymph node) , CD22 antigen ( expressed in particular in (0420 ] a transmembrane domain , appendix , lymph node, tonsil and spleen ), CD25 antigen [0421 ] an intracellular domain comprising a sequence ( expressed mainly in bladder and lymph node ) and MUC1 selected from the list consisting of SEQ ID NO : 1 antigen (expressed in kidney ). (human KI2L2 ), SEQ ID NO : 2 (human KI2L1) , SEQ [ 0404 ] Therefore , in one embodiment, the N - CAR of the ID NO : 3 (human FCG2B ) , SEQ ID NO : 4 (human invention comprises at least: KI2L3 ) , SEQ ID NO : 5 (human KI3L2 ) , SEQ ID NO :6 [0405 ] an extracellular domain comprising at least one (human KI2L4 ), SEQ ID NO : 7 (human KI3L1) , SEQ scFv from and an antibody binding specifically to an ID NO : 8 (human KIZLA ) , SEQ ID NO : 9 (human antigen selected from CD4 antigen , CD20 antigen , MILR1) , SEQ ID NO : 10 (human LIRB4 ) , SEQ ID CD22 antigen , CD25 antigen and /or MUC1 antigen ; or NO :11 (human LIRB3 ) , SEQ ID NO : 12 (human a combination thereof. KI3L3 ) , SEQ ID NO : 15 (human LIRB5 ) , SEQ ID [ 0406 ] a linker with a SEQ ID NO : 38 , SEQ ID NO : 39 NO : 16 (human LIRB2 ) , SEQ ID NO : 18 (human or SEQ ID NO : 40 ; and , FCRL4 ), SEQ ID NO :23 (human FCRL5 ) , SEQ ID [0407 ] a transmembrane domain NO : 24 ( human FCRL2 ) , SEQ ID NO : 25 (human [0408 ] an intracellular domain comprising a sequence FCRL1) , SEQ ID NO :27 (human FCRL3 ) , SEQ ID selected from the list consisting of SEQ ID NO : 33 NO : 28 (human MPZL1) , SEQ ID NO : 29 (human (human TR10D ), SEQ ID NO : 34 (human TR10A ) and PILRA ) or SEQ ID NO : 30 (human PVR ) . SEQ ID NO : 35 (human TR10B ) . [0422 ] In another embodiment, the N -CAR of the inven [ 0409 ] In another embodiment, the N -CAR of the inven tion comprises at least : tion comprises at least : 10423 ] an extracellular domain comprising at least one [0410 ] an extracellular domain comprising an at least scFv from an antibody binding specifically to an anti one scFv from an antibody binding specifically to an gen selected from CD4 antigen , CD20 antigen , CD22 antigen selected from CD4 antigen , CD20 antigen , antigen , CD25 antigen and / or MUC1 antigen ; CD22 antigen , CD25 antigen and / or MUC1 antigen ; [0424 ] a linker with a SEQ ID NO : 38 , SEQ ID NO :39 [0411 ] a linker with a SEQ ID NO : 38 , SEQ ID NO : 39 or SEQ ID NO : 40 ; and , or SEQ ID NO :40 ; and, [0425 ] an intracellular domain comprising a sequence [0412 ] an intracellular domain comprising a sequence of SEQ ID NO : 36 (CD200 receptor 1 ). with more than 80 % , preferably 90 % and more pref [0426 ] In another embodiment, N -CAR of the invention erably 95 % identity with SEQ ID NO : 33 (human comprises at least : TR10D ) , SEQ ID NO : 34 (human TR10A ) and SEQ ID [0427 ] an extracellular domain comprising at least one NO : 35 (human TR10B ) . scFv from an antibody binding specifically to an anti 10413 ] In another embodiment, the N -CAR of the inven gen selected from CD4 antigen , CD20 antigen , CD22 tion comprises at least : antigen , CD25 antigen and /or MUC1 antigen - a linker [ 0414 ] an extracellular domain comprising at least one with a SEQ ID NO : 38 , SEQ ID NO : 39 or SEQ ID scFv from an antibody binding specifically to an anti NO :40 ; and , gen selected from CD4 antigen , CD20 antigen , CD22 [0428 ] an intracellular domain comprising a sequence antigen , CD25 antigen and / or MUC1 antigen ; with more than more than 80 % , preferably 90 % and [0415 ] a linker with a SEQ ID NO : 38 , SEQ ID NO :39 more preferably 95 % identity with SEQ ID NO : 36 or SEQ ID NO : 40 ; and , (CD200 receptor 1 ) . [0416 ] an intracellular domain comprising a sequence (0429 ] For instance for the treatment of multiple myeloma with more than more than 80 % , preferably 90 % and (MM ) , when the antigen targeted by the P -CAR is CD38 , more preferably 95 % identity with SEQ ID NO : 1 said extra - binding domain of the N -CAR binds to at least (human KI2L2) , SEQ ID NO : 2 (human KI2L1) , SEQ one “ off- target" antigen expressed on healthy cells may be ID NO : 3 (human FCG2B ) , SEQ ID NO : 4 ( human chosen amongst an antigen selected from CD56 antigen KI2L3) , SEQ ID NO : 5 (human KI3L2 ) , SEQ ID NO : 6 (expressed on the surface of neurons, glia , skeletal muscle (human KI2L4 ), SEQ ID NO : 7 (human KI3L1) , SEQ and natural killer cells ) , CD205 antigen ( expressed on ID NO :8 (human KI2LA ) SEQ ID NO : 9 (human cortical thymic epithelial cells and by dendritic cell ( DC ) MILR1 ) , SEQ ID NO : 10 (human LIRB4 ) , SEO ID subsets ) , CD83 antigen ( expressed on activated lympho NO : 11 (human LIRB3 ) , SEQ ID NO : 12 (human cytes , Langerhans cells and interdigitating reticulum cells ) , KIZL3 ), SEQ ID NO : 15 (human LIRB5) , SEQ ID CD206 antigen ( expressed on the surface of macrophages NO : 16 (human LIRB2 ) , SEQ ID NO : 18 (human and dendritic cells , on the surface of skin cells such as FCRL4 ) , SEQ ID NO : 23 (human FCRL5 ) , SEQ ID human dermal fibroblasts and keratinocytes ); CD200 anti NO :24 (human FCRL2) , SEQ ID NO : 25 (human gen (expression on cells originating from the hematopoietic FCRL1) , SEQ ID NO : 27 (human FCRL3) , SEQ ID cells , activated T cells , endothelial neuronal cells and cells NO : 28 (human MPZL1) , SEQ ID NO :29 (human of the reproductive organs - ovaries and placental tropho PILRA ) or SEQ ID NO : 30 (human PVR ) . blasts - ); CD36 antigen ( expressed in adipocytes endothelial US 2017 /0296623 A1 Oct . 19 , 2017 25 cells and monocytes) ; RARRES1 antigen ( expressed of this KI2L3 ), SEQ ID NO : 5 (human KI3L2 ), SEQ ID NO : 6 gene upregulated by tazarotene as well as by retinoic acid (human KI2L4 ) , SEQ ID NO : 7 (human KI3L1) , SEQ receptors ) ID NO : 8 (human KIZLA ), SEQ ID NO : 9 (human [0430 ] For instance for the treatment ofmultiple myeloma MILR1 ) , SEQ ID NO : 10 (human LIRB4) , SEQ ID (MM ) , when the antigen targeted by the P -CAR is CD22 , NO : 11 (human LIRB3 ) , SEO ID NO : 12 (human said extra -binding domain of the N -CAR binds to at least KI3L3) , SEQ ID NO : 15 (human LIRB5) , SEQ ID one “ off - target” antigen expressed on healthy cells may be NO : 16 ( human LIRB2 ), SEQ ID NO : 18 (human chosen amongst an antigen selected from CD56 antigen FCRL4 ) , SEQ ID NO : 23 (human FCRL5 ) , SEQ ID ( expressed on the surface of neurons , glia , skeletal muscle NO :24 (human FCRL2 ) , SEQ ID NO : 25 (human and natural killer cells ) , CD205 antigen (expressed on FCRL1 ) , SEQ ID NO : 27 (human FCRL3) , SEQ ID cortical thymic epithelial cells and by dendritic cell (DC ) NO : 28 (human MPZL1) , SEQ ID NO :29 (human subsets ) , CD83 antigen ( expressed on activated lympho PILRA ) or SEQ ID NO : 30 (human PVR ). cytes , Langerhans cells and interdigitating reticulum cells ) , [ 0443 ] In another embodiment, the N -CAR of the inven CD206 antigen ( expressed on the surface of macrophages tion comprises at least : and dendritic cells , on the surface of skin cells such as [ 0444 ] an extracellular domain comprising at least one human dermal fibroblasts and keratinocytes ); CD200 anti scFv from an antibody binding specifically to an anti gen ( expression on cells originating from the hematopoietic gen selected from CD56 antigen , CD205 antigen , cells , activated T cells , endothelial neuronal cells and cells CD83 antigen , CD206 antigen ; CD200 antigen ; CD36 of the reproductive organs - ovaries and placental tropho antigen and /or RARRES1 antigen ; — a linker with a blasts - ) ; CD36 antigen ( expressed in adipocytes endothelial SEQ ID NO : 38 , SEQ ID NO :39 or SEQ ID NO : 40 ; cells and monocytes) ; RARRES1 antigen ( expressed of this and , gene upregulated by tazarotene as well as by retinoic acid [0445 ] an intracellular domain comprising a sequence receptors ) selected from the list consisting of SEQ ID NO : 1 [ 0431] Therefore , in one embodiment, the N - CARs of the (human KI2L2 ), SEQ ID NO : 2 (human KI2L1 ), SEQ invention comprises at least: ID NO : 3 (human FCG2B ) , SEQ ID NO : 4 (human [0432 ] an extracellular domain comprising at least one KI2L3 ) , SEQ ID NO : 5 (human KI3L2 ) , SEQ ID NO : 6 scFv from an antibody binding specifically to an anti (human KI2L4 ) , SEQ ID NO : 7 (human KI3L1) , SEQ gen selected from CD56 antigen , CD205 antigen , ID NO : 8 (human KIZLA ) , SEQ ID NO : 9 (human CD83 antigen , CD206 antigen ; CD200 antigen ; CD36 MILR1 ) , SEO ID NO : 10 (human LIRB4 ) , SEO ID antigen and / or RARRES1 antigen , NO :11 (human LIRB3 ), SEQ ID NO :12 (human [ 0433 ] a linker with a SEQ ID NO :38 , SEQ ID NO : 39 KI3L3) , SEQ ID NO : 15 (human LIRB5) , SEQ ID or SEQ ID NO :40 ; and , NO : 16 (human LIRB2) , SEQ ID NO : 18 (human [0434 ] an intracellular domain comprising a sequence FCRL4 ) , SEQ ID NO :23 (human FCRL5 ) , SEQ ID with more than 80 % , preferably 90 % and more pref NO : 24 (human FCRL2 ) , SEQ ID NO : 25 (human erably 95 % identity with SEQ ID NO : 33 (human FCRL1 ), SEQ ID NO :27 (human FCRL3 ), SEQ ID TR10D ) , SEQ ID NO : 34 (human TR10A ) and SEQ ID NO : 28 (human MPZL1) , SEQ ID NO : 29 (human NO : 35 (human TR10B ) . PILRA ) or SEQ ID NO : 30 (human PVR ). [0435 ] In one embodiment, the N -CAR of the invention [ 0446 ] In another embodiment, the N -CAR of the inven comprises at least: tion comprises at least : [0436 ] an extracellular domain comprising at least one [0447 ] an extracellular domain comprising at least one scFv from an antibody binding specifically to an anti scFv from an antibody binding specifically to an anti gen selected from CD56 antigen , CD205 antigen , gen selected from CD56 antigen , CD205 antigen , CD83 antigen , CD206 antigen ; CD200 antigen ; CD36 CD83 antigen , CD206 antigen ; CD200 antigen ; CD36 antigen and /or RARRES1 antigen ; antigen and / or RARRES1 antigen ; [ 0437 ] a linker with a SEQ ID NO : 38 , SEQ ID NO : 39 [ 0448 ] a linker with a SEQ ID NO : 38 , SEQ ID NO : 39 or SEQ ID NO : 40 ; and , or SEQ ID NO :40 ; and , [ 0438 ] an intracellular domain comprising a sequence 104491 an intracellular domain comprising a sequence selected from the list consisting of SEQ ID NO : 33 with more than more than 80 % , preferably 90 % and (human TR10D ) , SEQ ID NO : 34 (human TR10A ) or more preferably 95 % identity with SEQ ID NO : 36 SEQ ID NO : 35 (human TR10B ) , (CD200 receptor 1 ) . [ 0439 ] In another embodiment, the N - CAR of the inven [ 0450 ] In another embodiment, the N -CAR of the inven tion comprises at least: tion comprises at least : [ 0440 ] an extracellular domain comprising at least one [ 0451] an extracellular domain comprising at least one scFv from an antibody binding specifically to an anti scFv from antibodies binding specifically to an antigen gen selected from CD56 antigen , CD205 antigen , selected from CD56 antigen , CD205 antigen , CD83 CD83 antigen , CD206 antigen ; CD200 antigen ; CD36 antigen , CD206 antigen ; CD200 antigen ; CD36 antigen antigen and /or RARRES1 antigen ; and / or RARRES1 antigen ; [0441 ] a linker with a SEQ ID NO :38 , SEQ ID NO :39 [0452 ] a linker with a SEQ ID NO :38 , SEQ ID NO :39 or SEQ ID NO :40 ; and , or SEQ ID NO :40 ; and , [0442 ] an intracellular domain comprising a sequence [0453 ] an intracellular domain comprising a sequence with more than more than 80 % , preferably 90 % and of SEQ ID NO : 36 (CD200 receptor 1) . more preferably 95 % identity with SEQ ID NO : 1 [0454 ] For instance for the treatment of chronic lympho (human KI2L2) , SEQ ID NO : 2 human KI2L1) , SEQ cytic leukemia (CLL ) , when the antigen targeted by the ID NO :3 (human FCG2B ) , SEQ ID NO :4 (human P -CAR is CS1, said extra -binding domain of the N -CAR US 2017 /0296623 A1 Oct . 19 , 2017 binds to " off -target ” antigens expressed on healthy cells that gen selected from troponin C antigen , beta - 1 integrin may be chosen amongst troponin C antigen (expressed in antigen ; CCKBR antigen ; GALR1 antigen and /or heart ) ; beta - 1 integrin antigen ( expressed in endothelial cells CUBN antigen ; and fibroblasts , intestine , colon , testis , ovary , thymus, spleen [ 0469 ] a linker with a SEQ ID NO : 38 , SEQ ID NO : 39 and prostate ) ; CCKBR antigen ( expression in stomach , or SEQ ID NO :40 ; and , pancreas , brain and gallbladder ) ; GALR1 antigen ( expressed [ 0470 ] an intracellular domain comprising a sequence in adrenal gland ) ; and CUBN antigen ( expressed in kidney selected from the list consisting of SEQ ID NO : 1 and small intestine ) . (human KI2L2 ), SEQ ID NO : 2 (human KI2L1) , SEQ [0455 ] Therefore, in one embodiment, the N -CAR of the ID NO : 3 (human FCG2B ), SEQ ID NO : 4 (human invention may comprise at least : KI2L3 ) , SEQ ID NO : 5 (human KI3L2 ) , SEQ ID NO : 6 [0456 ] an extracellular domain comprising at least one (human KI2L4 ), SEQ ID NO : 7 (human KI3L1) , SEQ scFv from an antibody binding specifically to an anti ID NO : 8 (human KIZLA ), SEQ ID NO : 9 (human gen selected from troponin C antigen ; beta - 1 integrin MILR1) , SEQ ID NO : 10 (human LIRB4) , SEQ ID antigen ; CCKBR antigen ; GALR1 antigen and CUBN NO : 11 (human LIRB3 ), SEQ ID NO : 12 (human antigen ; KI3L3 ) , SEQ ID NO : 15 (human LIRB5) , SEQ ID [0457 ] a linker with a SEQ ID NO : 38, SEQ ID NO :39 NO : 16 (human LIRB2) , SEQ ID NO : 18 (human or SEQ ID NO : 40 ; and , FCRL4) , SEQ ID NO :23 ( human FCRL5) , SEQ ID [0458 ] an intracellular domain comprising a sequence NO : 24 (human FCRL2 ) , SEQ ID NO : 25 (human with more than 80 % , preferably 90 % and more pref erably 95 % identity with SEQ ID NO : 33 (human FCRL1) , SEQ ID NO : 27 (human FCRL3 ) , SEQ ID TR10D ) , SEQ ID NO : 34 (human TR10A ) or SEQ ID NO : 28 (human MPZL1) , SEQ ID NO : 29 (human NO : 35 ( human TR10B ) . PILRA ) or SEQ ID NO : 30 (human PVR ) . [0459 ] In one embodiment , the N -CAR of the invention [0471 ] In another embodiment, the N -CAR of the inven comprises at least: tion comprises at least : [0460 ] an extracellular domain comprising at least one [0472 ] an extracellular domain comprising at least one scFv from an antibody binding specifically to an anti scFv from an antibody binding specifically to an anti gen selected from troponin C antigen beta - 1 integrin gen selected from troponin C antigen , beta - 1 integrin antigen ; CCKBR antigen ; GALR1 antigen and / or antigen ; CCKBR antigen ; GALR1 antigen and /or CUBN antigen ; CUBN antigen ; [0461 ] a linker with a SEQ ID NO : 38, SEQ ID NO :39 [ 0473 ] a linker with a SEQ ID NO : 38 , SEQ ID NO : 39 or SEQ ID NO : 40 ; and , or SEQ ID NO :40 ; and , [0462 ] an intracellular domain comprising a sequence [ 04741 an intracellular domain comprising a sequence selected from the list consisting of SER ID NO : 33 with more than 80 % , preferably 90 % and more pref (human TR10D ) , SEQ ID NO : 34 (human TR10A ) or erably 95 % identity with SEQ ID NO : 36 (CD200 SEQ ID NO : 35 (human TR10B ) . receptor 1 ) . [0463 ] In another embodiment, the N -CAR of the inven [0475 ] In another embodiment, the N -CAR of the inven tion comprises at least: tion comprises at least : 10464 ) an extracellular domain comprising at least one [0476 ] an extracellular domain comprising at least one scFv from an antibody binding specifically to an anti scFv from an antibody binding specifically to an anti gen selected from troponin C antigen beta - 1 integrin gen selected from troponin C antigen (; beta - 1 integrin antigen ; CCKBR antigen ; GALR1 antigen and /or antigen ; CCKBR antigen ; GALR1 antigen and /or CUBN antigen ; CUBN antigen ; [ 0465 ] a linker with a SEQ ID NO : 38 , SEQ ID NO : 39 or SEQ ID NO :40 ; and, [ 0477 ] a linker with a SEQ ID NO : 38 , SEQ ID NO : 39 [ 0466 ] an intracellular domain comprising a sequence or SEQ ID NO :40 ; and , with more than more than 80 % , preferably 90 % and [0478 ] an intracellular domain comprising a sequence more preferably 95 % identity with SEQ ID NO : 1 of SEQ ID NO :36 ( CD200 receptor 1) . ( human KI2L2 ) , SEQ ID NO : 2 human KI2L1) , SEO [ 0479 ] For instance for the treatment of chronic lympho ID NO : 3 (human FCG2B ) , SEQ ID NO : 4 (human cytic leukemia (CLL ) or a solid tumor, when the antigen KI2L3) , SEQ ID NO : 5 ( human KI3L2) , SEQ ID NO :6 targeted by the P -CAR is ROR1, said extra -binding domain ( human KI2L4) , SEQ ID NO : 7 (human KI3L1) , SEQ of the N -CAR binds to at least one “ off- target " antigen ID NO : 8 ( human KIZLA ) , SEQ ID NO : 9 (human expressed on healthy cells selected from the list consisting MILR1) , SEQ ID NO : 10 (human LIRB4 ), SEQ ID of troponin C antigen ( expressed in heart ) ; beta - 1 integrin NO : 11 (human LIRB3) , SEQ ID NO : 12 (human antigen (expressed in endothelial cells and fibroblasts , intes KI3L3 ) , SEO ID NO : 15 (human LIRB5 ) , SEO ID tine , colon , testis , ovary, thymus, spleen and prostate) ; NO : 16 (human LIRB2 ) , SEQ ID NO : 18 (human CCKBR antigen (expression in stomach , pancreas, brain and FCRL4 ) , SEQ ID NO : 23 (human FCRL5 ) , SEQ ID gallbladder ) ; GALR1 antigen ( expressed in adrenal gland ) NO :24 (human FCRL2) , SEQ ID NO : 25 (human or MUC1 antigen (expressed in kidney ). FCRL1) , SEQ ID NO :27 (human FCRL3 ) , SEQ ID [0480 ] Therefore , in one embodiment, the N -CAR of the NO : 28 (human MPZL1) , SEQ ID NO :29 (human invention comprises at least: PILRA ) or SEQ ID NO : 30 (human PVR ) . [ 0481 ] an extracellular domain comprising at least one [0467 ] In another embodiment, the N -CAR of the inven scFv from an antibody binding specifically to an anti tion comprises at least : gen selected from troponin C antigen (; beta - 1 integrin [0468 ] an extracellular domain comprising at least one antigen ; CCKBR antigen ; GALR1 antigen and /or scFv from an antibody binding specifically to an anti MUC antigen ; US 2017 /0296623 A1 Oct . 19 , 2017 27 [ 0482 ] a linker with a SEQ ID NO : 38 , SEQ ID NO : 39 KI3L3 ) , SEQ ID NO : 15 (human LIRB5 ) , SEQ ID or SEQ ID NO :40 ; and , NO : 16 (human LIRB2) , SEQ ID NO : 18 (human [0483 ] an intracellular domain comprising a sequence FCRL4 ) , SEQ ID NO : 23 (human FCRL5 ) , SEQ ID with more than 80 % , preferably 90 % and more pref NO : 24 (human FCRL2) , SEQ ID NO : 25 (human erably 95 % identity with SEQ ID NO : 33 (human FCRL1) , SEQ ID NO : 27 ( human FCRL3 ) , SEQ ID TR10D ) , SEQ ID NO : 34 (human TR10A ) and SEQ ID NO : 28 (human MPZL1) , SEQ ID NO :29 (human NO : 35 (human TR10B ) . PILRA ) or SEQ ID NO : 30 (human PVR ) . 104841 In one embodiment, the N -CAR of the invention [0496 ] In another embodiment, the N -CAR of the inven comprises at least : tion comprises at least : [0485 ] an extracellular domain comprising at least one scFv from an antibody binding specifically to an anti [ 0497 ] an extracellular domain comprising at least one gen selected from troponin C antigen (; beta - 1 integrin scFv from an antibody binding specifically to an anti antigen ; CCKBR antigen ; GALR1 antigen and /or gen selected from troponin C antigen ( ; beta - 1 integrin MUC antigen ; antigen ; CCKBR antigen ; GALR1 antigen and /or [ 0486 ] a linker with a SEQ ID NO : 38 , SEQ ID NO : 39 MUC antigen . or SEQ ID NO : 40 ; and , [ 0498 ] In other embodiments , the N - CAR of the present [0487 ] an intracellular domain comprising a sequence invention is a transmembrane polypeptide containing at selected from the list consisting of SEQ ID NO : 33 least: (human TR10D ) , SEQ ID NO : 34 (human TR10A ) or [0499 ] an extracellular binding domain ; SEQ ID NO : 35 (human TR10B ) [ 0500 ] an intracellular domain comprising an inhibitory [0488 ] In another embodiment, N -CARs of the invention transduction domain , may comprise at least : [0489 ] an extracellular domain comprising at least one [ 0501 ] wherein said inhibitory transduction domain of scFv from an antibody binding specifically to an anti intracellular domain is used alone, fused to a separately gen selected from troponin C antigen ( ; beta - 1 integrin chosen transmembrane domain , optionally , the latter being Tiinantigen ; CCKBR antigen ; GALR1 antigen and / or fused to the extracellular binding domain by a hinge . MUC antigen ; [ 0502 ] Transmembrane Domain [0490 ] a linker with a SEQ ID NO : 38, SEQ ID NO :39 [0503 ] In one embodiment, the transmembrane domain or SEQ ID NO : 40 ; and , comprises the transmembrane region ( s ) of the alpha , beta or [0491 ] an intracellular domain comprising a sequence zeta chain of the T -cell receptor, PD - 1 , 4 - 16B , OX40 , ICOS, with more than more than 80 % , preferably 90 % and CTLA - 4 , LAG3 , 2B4 , BTLA4 , TIM - 3 , TIGIT , SIRPA , more preferably 95 % identity with SEO ID NO : 1 CD28 , CD3 epsilon , CD45, CD4 , CD5 , CD , CD9, CD16 , (human KI2L2 ) , SEQ ID NO : 2 human KI2L1) , SEQ CD22 , CD33 , CD37 , CD64 , CD80 , CD86 , CD134 , CD137 ID NO : 3 (human FCG2B ) , SEQ ID NO : 4 (human or CD154 . KI2L3 ) , SEQ ID NO : 5 (human KI3L2 ) , SEQ ID NO : 6 [0504 ] The distinguishing features of appropriate trans (human KI2L4 ), SEQ ID NO : 7 (human KI3L1) , SEQ membrane domains comprise the ability to be expressed at ID NO : 8 ( human KIZLA ) , SEQ ID NO : 9 (human the surface of a cell , preferably in the present invention an MILR1 ) , SEQ ID NO : 10 (human LIRB4 ), SEQ ID immune cell, in particular lymphocyte cells or Natural killer NO : 11 ( human LIRB3 ) , SEQ ID NO : 12 (human (NK ) cells , and to interact together for directing cellular KI3L3 ), SEQ ID NO : 15 (human LIRB5 ) , SEQ ID response of immune cell against a predefined target cell. The NO : 16 (human LIRB2 ) , SEO ID NO : 18 (human transmembrane domain can be derived either from a natural FCRL4 ) , SEQ ID NO : 23 ( human FCRL5 ) , SEQ ID or from a synthetic source . The transmembrane domain can NO : 24 (human FCRL2 ) , SEQ ID NO : 25 (human be derived from any membrane -bound or transmembrane FCRL1) , SEQ ID NO :27 (human FCRL3) , SEQ ID protein . As non - limiting examples , the transmembrane poly NO : 28 ( human MPZL1) , SEQ ID NO :29 (human peptide can be a subunit of the T cell receptor such as a B , PILRA ) or SEQ ID NO : 30 (human PVR ) . ? or ? , polypeptide constituting CD3 complex , IL2 recep [0492 ] In another embodiment, the N -CAR of the inven tor p55 ( a chain ) , p75 ( ß chain ) or ? chain , subunit chain tion comprises at least : of Fc receptors , in particular Fc ? receptor III or CD pro [0493 ] an extracellular domain comprising at least one teins . Alternatively the transmembrane domain can be syn scFv from an antibody binding specifically to an anti thetic and can comprise predominantly hydrophobic resi gen selected from troponin C antigen ( ; beta - 1 integrin dues such as leucine and valine . In a preferred embodiment antigen ; CCKBR antigen ; GALR1 antigen and/ or said transmembrane domain is derived from the human CD8 MUC antigen ; alpha chain ( e . g . NP _ 001139345 . 1 ) . Said transmembrane [ 0494 ] a linker with a SEQ ID NO : 38 , SEQ ID NO : 39 domain can also be a CD8 transmembrane domain ( alpha or SEQ ID NO : 40 ; and , and beta chains) . Said Transmembrane domain can be engi [04951 an intracellular domain comprising a sequence neered to create obligated hetero or homodimers . In particu selected from the list consisting of SER ID NO : 1 lar embodiment said CARs can comprise transmembrane (human KI2L2) , SEQ ID NO : 2 (human KI2L1) , SEQ domains or intracellular domains which can only dimerize ID NO : 3 ( human FCG2B ) , SEO ID NO : 4 (human after ligand recognition . Another example of transmembrane KI2L3) , SEQ ID NO : 5 ( human KI3L2 ), SEQ ID NO : 6 domain can be NKG2- D receptor. NKG2D ( natural killer (human KI2L4 ), SEQ ID NO : 7 (human KI3L1) , SEQ cell group 2D ) is a C - type lectin - like receptor expressed on ID NO : 8 ( human KIZLA ) , SEQ ID NO : 9 (human NK cells, yo - TcR + T cells , and CD8 + aß - TcR + T cells (Bauer , MILR1) , SEQ ID NO : 10 (human LIRB4 ) , SEQ ID Groh et al ., 1999, Science 285 (5428 ): 727 -9 . NKG2D is NO : 11 (human LIRB3 ) , SEQ ID NO : 12 (human associated with the transmembrane adapter protein DAP10 US 2017 /0296623 A1 Oct . 19 , 2017 (Wu , Song et al. 1999 , Science 285 (5428 ) :730 - 2 ), whose [0511 ] In a more preferred embodiment, the hinge of the cytoplasmic domain binds to the p 85 subunit of the PI- 3 N -CAR is an IgG1 hinge or a CD8 alpha hinge . kinase . [0512 ] The term “ stalk region ” (also named hinge region ) [0505 ] Another example of transmembrane domain can be used herein generally means any oligo - or polypeptide that a receptor tyrosine kinase . Receptor tyrosine kinase are cell functions to link the transmembrane domain to the extra surface receptors involved in different critical cellular regu cellular ligand -binding domain . In particular, stalk region latory process including cell proliferation , cell differentia are used to provide more flexibility and accessibility for the tion , cell survival, cell migration , as well as cell cycle extracellular ligand - binding domain . A stalk region may control. Receptor tyrosine kinase comprises an extracellular comprise up to 300 amino acids, preferably 10 to 100 amino domain , a single transmembrane helix and an intracellular acids and most preferably 25 to 50 amino acids. Stalk region domain comprising tyrosine kinase function that is most of may be derived from all or part of naturally occurring time autoregulated by additional carboxy - terminal and jux molecules , such as from all or part of the extracellular region tamembrane domains . Activation of receptor tyrosine kinase of CD , CD4 , CD28 or RTK , or from all or part of an is generally elicited by ligand -mediated dimerization . antibody constant region . Alternatively the stalk region may Thanks to their bivalence , growth hormone ligand has the capacity to simultaneously interact with two receptor mono be a synthetic sequence that corresponds to a naturally mers and promotes dimerization . Such dimerization induces occurring stalk sequence , or may be an entirely synthetic the activation of intracellular kinase domains through con stalk sequence . formational changes followed by trans -phosphorylation of (0513 ]. The present invention encompasses a recombinant different tyrosines located within their intracellular domain . DNA construct comprising sequences encoding an N -CAR The different phosphotyrosines generated eventually serve as defined above, wherein the N -CAR comprises an extra as docking site for the recruitment of downstream signaling cellular domain such as an antibody fragment that binds partners that activate the cellular regulatory pathways . Said specifically to an off - tumor antigen , and wherein the CAR can comprise the extracellular domain , transmem sequence of the extracellular domain is contiguous with and brane, and/ or the intracellular domain of a receptor tyrosine in the same reading frame as a nucleic acid sequence kinase , preferably selected from the group consisting of encoding a transmembrane domain and an intracellular TrkA , C -Kit , FGFR and EGFR / Erb . Said tyrosine kinase domain . An exemplary N -CAR construct may comprise an transmembrane domain and / or intracellular domain can be optional leader sequence , an extracellular off - tissue antigen linked to an extracellular ligand binding domain and intra binding domain , a hinge , a transmembrane domain , and an cellular domain according to the present invention . Said intracellular inhibitory signaling domain . engineered cells may comprise different N - and P - CAR [0514 ] According to one preferred embodiment, a hinge comprising different transmembrane domains. according to the invention comprises the a sequence from [0506 ] Said transmembrane domain can also be an integ IgG1 or from CD8 alpha , preferably of SEQ ID NO . 51 and rin . Integrins are heterodimeric integral membrane proteins composed of a 2 and ? ? chains which combined together 50 . form the LFA - 1 ( integrin lymphocyte function - associated [0515 ] Engineered Immune Cells antigen - 1 ) which is expressed on all leukocytes. LFA - 1 [0516 ] In one aspect the present invention provides an plays a central role in leukocyte intercellular adhesion immune cell comprising at least one N -CAR according to through interactions with its ligand , ICAMs 1 - 3 ( intercellu the invention ( as described above ) . lar adhesion molecules 1 through 3 ) , and also it has an [ 0517 ] In one aspect the present invention provides an important role in lymphocyte co - stimulatory signaling immune cell comprising at least one N - CAR according to (Chen and Flies 2013 , Nat Rev Immunol 13 ( 4 ) : 227 -42 ) . The the invention (as described above ) and at least one P -CAR , molecular details of the binding of LAF - 1 to its immuno according to the invention . globulin ICAM - 1 are quite known allowing a careful engi [0518 ] In one aspect of the invention , an isolated immune neering ofLAF - 1 binding site . The affinity of ? , domain for ICAM - 1 is regulated by the displacement of its C - terminal cell comprises a P - CAR comprising: helix which is conformational linked to alterations of spe [ 0519 ] an extracellular domain comprising an antigen cific loops in LAF - 1 . The active and low conformations binding domain ; differ of 500 and 10 ,000 folds. It is also interesting to note [ 0520 ] a transmembrane domain ; that two types of antagonists are known for LFA - 1 and their [ 0521 ] an intracellular domain ; mechanism of action is known . Integrin cell surface adhe [ 0522 ] and an N - CAR as described previously . sion receptors can transmit a signal from the outside to [ 0523] The present invention encompasses an immune cell inside but also vice - versa . There are cytoskeletal proteins as comprising a single chain (sc ) or a multi chain (mc ) N -CAR Talin which binds to the integrin tail LFA - 1 to transfer a and a sc P - CAR . message from inside to outside . [0524 ] The present invention encompasses an immune cell [0507 ] According to one embodiment, the transmembrane comprising a single chain ( sc ) or a multi chain (mc ) N - CAR domain comprises the transmembrane region of PD - 1 or the and a mc P -CAR . transmembrane region ( s ) of CD8 alpha . [0525 ]. The present invention encompasses an immune cell [0508 ] According to one preferred embodiment, the trans comprising a single chain ( sc ) N -CAR and a sc P -CAR . membrane domain comprises the transmembrane region of 10526 ] The present invention encompasses an immune cell CD8 alpha . In one aspect of the invention , the transmem comprising a multi chain (mc ) N -CAR and a sc P -CAR . brane domain is attached to the extracellular domain of the [ 0527 ] The present invention encompasses an immune cell N -CAR via a hinge . comprising a multi chain (mc ) N -CAR and a mc P -CAR . [0509 ] Hinge The present invention also relates to isolated cells or cell [0510 ] In a preferred embodiment, in the hinge of the lines susceptible to be obtained by said method to engineer N -CAR is a human immunoglobulin hinge . cells . US 2017 /0296623 A1 Oct . 19 , 2017 29 [0528 ] The present invention also relates to isolated cells [ 0539 ] According to one embodiment , the antigen to or cell lines susceptible to be obtained by a method to which the antigen binding domain of the P - CAR binds is engineer cells according to the present invention . CD20 and the antigen to which the antigen binding domain [0529 ] Said immune cell refers to a cell of hematopoietic of the N -CAR binds is an antigen selected from the list origin functionally involved in the initiation and / or execu consisting of CD56 , CD205 , CD83 , CD206 , CD200 and tion of innate and/ or adaptive immune response . Said CD36 . immune cell according to the present invention can be 10540 ] According to one embodiment , the antigen to derived from a stem cell . The stem cells can be adult stem which the antigen binding domain of the P -CAR binds is cells , non - human embryonic stem cells , more particularly PCMA and the antigen to which the antigen binding domain non - human stem cells , cord blood stem cells , progenitor of the N -CAR binds is an antigen selected from the list cells , bone marrow stem cells , induced pluripotent stem consisting of CD56 , CD205, CD83, CD206 , CD200 and cells , totipotent stem cells or hematopoietic stem cells . CD36 . Representative human cells are CD34 + cells . Said isolated 0541 ] According to another embodiment, the antigen cell can also be a dendritic cell, killer dendritic cell , a mast binding domain of the P -CAR binds is CS1 and the antigen cell , a NK - cell, a B - cell or a T cell. Said isolated cell may to which the antigen binding domain of the N -CAR binds is comprise a population of N - CARs and CARs each one an antigen selected from the list consisting of troponin C , comprising different extracellular ligand binding domains . beta - 1 integrin , CCKBR , GALR1 or CUBN . In particular, said isolated cell comprises exogenous poly 10542 ]. According to another embodiment, the antigen to nucleotide sequence encoding N -CAR and P -CAR . which the antigen binding domain of the P -CAR binds is [ 0530 ] In one preferred embodiment, said isolated cell CD123 and the antigen to which the antigen binding domain comprising at least one N -CAR and one CAR as described of the N -CAR binds is an antigen selected from the list above is a T - cell . consisting of CD4 , CD20 , CD22 , CD25 or MUC1. [0531 ] In a more preferred embodiment, said isolated cell [0543 ] According to another embodiment, the antigen to comprising at least one N -CAR and one CAR as described which the antigen binding domain of the P -CAR binds is above is a human T - cell . ROR1 and the antigen to which the antigen binding domain [0532 ] In a preferred embodiment, isolated immune cell is of the N -CAR binds is an antigen selected from the list selected from the group consisting of inflammatory T - lym consisting of troponin C , beta - 1 integrin , CCKBR , GALR1 phocytes , cytotoxic T- lymphocytes , regulatory T- lympho or MUC1. cytes or helper T -lymphocytes . [0544 ] Positive Chimeric Antigen Receptor (P -CAR ) [0533 ] Said cell may be derived from the group consisting [0545 ] The present invention relates to “ logical NOT” of CD4 + T - lymphocytes and CD8 + T - lymphocytes. Prior to gates that involve, beside the above described N -CAR , at expansion and genetic modification of the cells of the least one P - CAR which enable the engineered immune cell invention , a source of cells can be obtained from a subject to trigger the destruction of tumoral targeted cells . through a variety of non - limiting methods . Cells can be obtained from a number of non - limiting sources , including [0546 ] The P -CAR used within the scope of the invention peripheral blood mononuclear cells, bone marrow , lymph can be a single - chain or a multi - chain CAR . node tissue , cord blood , thymus tissue , tissue from a site of [0547 ] In one embodiment, the P -CAR is a single CAR ; it infection , ascites , pleural effusion , spleen tissue , and tumors . comprises one transmembrane polypeptide comprising at In certain embodiments of the present invention , any num least one extracellular ligand - binding domain and one extra ber of T cell lines available and known to those skilled in the cellular domain comprising a signal - transducing domain . art, may be used . [0548 ] In some embodiments , the immune cell comprises [ 0534 ] In one embodiment, said isolated immune cells are a multi - chain P -CAR as defined in WO2014 / 039523 which recovered from a healthy donor. is incorporated herein by reference in its entirety [0535 ] In another embodiment, said isolated immune cells [0549 ] By multi -chain CAR is meant a CAR structure that are recovered from a patient diagnosed with cancer or from comprises different polypeptides such as at least ( 1 ) a a patient diagnosed with an infection . transmembrane polypeptide which comprises at least one [0536 ] Said cells may part of a mixed population of cells extracellular ligand binding domain ; and ( 2 ) a transmem which present different phenotypic characteristics . In the brane polypeptide comprising at least one transduction scope of the present invention is also encompassed a cell line domain such that said at least two polypeptides assemble obtained from a transformed T - cell according to the method together to form a functional multi- chain Chimeric Antigen previously described . Receptor (WO2014039523 ) . [ 0537 ) According to one embodiment, the antigen to [0550 ] In another embodiment, this P -CAR is a multichain which the antigen binding domain of the P -CAR binds is CAR such as described in WO2014039523 , it comprises at CD38 and the antigen to which the antigen binding domain least : of the N -CAR binds is an antigen selected from the list [ 0551] one transmembrane polypeptide comprising at consisting of CD56 , CD205 , CD83 , CD206 , CD200 and least one extracellular ligand -binding domain and ; CD36 . [0552 ] one transmembrane polypeptide comprising at [0538 ] According to one embodiment, the antigen to least one signal- transducing domain . which the antigen binding domain of the P -CAR binds is [0553 ] For instance , said multi -chain CAR can comprise CD19 and the antigen to which the antigen binding domain at least two of the following components : of the N -CAR binds is an antigen selected from the list [0554 ] a ) one polypeptide comprising the transmembrem consisting of CD56 , CD205 , CD83 , CD206 , CD200 and brane domain of FcsRI alpha chain fused to an extracellular CD36 . ligand -binding domain , US 2017 /0296623 A1 Oct . 19 , 2017 30 [ 0555 ] b ) one polypeptide comprising a part of N - and vation of at least one of the normal functions of the engi C -terminal cytoplasmic tail fused to the transmembrane neered immune cell . For example , the function of a T cell domain of a FcRI beta chain , and/ or can be a cytolytic activity or helper activity including the [0556 ] c ) two additional polypeptides comprising each secretion of cytokines . Thus , the term " signaling protein ” one part of an intracytoplasmic tail and /or the transmem refers to a protein which transduces the transmitter domain brane domain of FcRI gamma chain , function signal and directs the cell to perform a specialized [ 0557] whereby these different polypeptides multimerize function . In a particular embodiment, said signaling domain together spontaneously to form dimeric , trimeric or tetra can be a signaling protein . Transmission of the signals can meric CARs. result from : protein /protein interactions, protein /DNA inter [0558 ] In a preferred embodiment said chain are not action , protein /RNA interaction , protein / small molecule covalently linked . interaction , post translational protein modification , confor [0559 ] Example of a tetrameric P - CARs are illustrated in mational change, subcellular relocalization . FIG . 3 of WO2013176915 and different versions of multi 10566 ] The signaling protein can activate a gene in the chain P -CARs are represented in FIG . 4 of WO2013176915 . nucleus . Examples of signaling protein can be members of Such P -CAR can be expressed in a T - cell obtained using the NFAT transcription factor family which are inducible factor above disclosed method together with a N -CAR according that could bind the intereukin - 2 promoter in activated T to the present disclosure to obtain a T - cell according to the cells . The regulation of NFAT proteins involves metabolites invention . and proteins such as calcium , calcineurin and Homer scaf [ 0560 ] In some embodiment the invention relates to an folding proteins . Said signaling protein can be an activated immune cell comprising a N - CAR as defined herein and a engineered form of NFAT avoiding regulation by calcineurin P -CAR as defined in any of U .S . Pat. No . 7 ,446 , 190 , and Homer proteins . Said signaling protein can be a NF -KB WO2008 / 121420 , U . S . Pat. No . 8 ,252 , 592 , US20140024809 , WO2012 /079000 , WO2014153270 , engineered to avoid sequestration in the cytoplasm by Il2b WO2012 /099973 , WO2014 /011988 , WO2014 /011987 , allowing activation of T cells . Said signaling protein can WO2013 /067492 , WO2013 /070468 , WO2013 /040557 , also be the expression of the three IKK subunits (IKKA , WO2013 / 126712 , WO2013 / 126729 , WO 2013 / 126726 , IKKB , IKKy) . Reconstituted IKK complex activated NF WO2013/ 126733 , U . S . Pat. No . 8 , 399, 645 , BB pathway, by triggering the ubiquitination of the IKB . US20130266551 , US20140023674 , WO2014039523 , U . S . Also the activation of the JNK signaling could be triggered Pat. No. 7 ,514 ,537 , U . S . Pat . No . 8 ,324 , 353 , WO2010 / through the direct expression of signaling protein AP - 1 025177 , U . S . Pat. No . 7 , 446 , 179 , WO2010 / 025177 , ( transcription factor ) . Said signaling protein can be an WO2012 /031744 , WO2012 / 136231A1, WO2012 / engineered transcription activator like effector ( TALE) bind 050374A2 , WO2013074916 , WO / 2009 /091826A3 , ing domain that will specifically target and activate tran WO2013 /176915 or WO /2013 /059593 . scription of the same gene as for the NFAT and NF- kb . [0561 ] The transmembrane domain of the P -CAR [ 0567 ] According to the invention , said signaling protein responds to similar criteria that the one explained previously can inhibit a signaling pathway through protein -protein for the N -CAR . Idem for the extracellular ligand -binding interaction or can activate a gene in the nucleus to inhibit a domain of P - CAR , excepted the difference of specificity signaling pathway . Said signaling protein can be vaccinia H1 towards its antigen target as presented above. related proteins (VHR ) a member of the mitogen -activated [ 0562 ] A preferred TM is from CD8 alpha, more prefer protein kinase phosphatases (MKPs ) family which dephos ably of SEQ ID NO . 50 phorylates and inactivates an extracellular signal regulated [0563 ] Example of a tetrameric P -CARs are illustrated in kinases (ERK ) signaling proteins. FIG . 3 of W02013176915 and different versions of multi chain P -CARs are represented in FIG . 4 of WO2013176915 . [0568 ] According to the invention , a signal transducing Such P -CAR can be expressed in a T - cell obtained using the domain for use in a P -CAR can be the cytoplasmic above disclosed method together with a N -CAR according sequences of the T cell receptor and co -receptors that act in to the present disclosure to obtain a T -cell according to the concert to initiate signal transduction following antigen invention . receptor engagement, as well as any derivate or variant of [ 0564 ] In some embodiment the invention relates to an these sequences and any synthetic sequence that has the immune cell comprising a N -CAR as defined herein and a same functional capability . Signal transduction domain may P -CAR as defined in any of U . S . Pat. No . 7 ,446 , 190 , comprise two distinct classes of cytoplasmic signaling WO2008 / 121420 , U . S . Pat. No . 8 ,252 , 592 , sequence, those that initiate antigen - dependent primary acti US20140024809 , WO2012 /079000 , WO2014153270 , vation , and those that act in an antigen - independent manner WO2012 /099973 , WO2014 /011988 , WO2014 /011987 , to provide a secondary or co - stimulatory signal . WO2013 / 067492 , WO2013 /070468 , WO2013 / 040557 , [0569 ] Primary cytoplasmic signaling sequence can com WO2013 / 126712 , WO2013 / 126729 , WO 2013 / 126726 , prise signaling motifs which are known as immunoreceptor WO2013 / 126733 , U . S . Pat. No . 8 , 399 ,645 , tyrosine -based activation motifs of ITAMs. ITAMs are well US20130266551 , US20140023674 , WO2014039523 , U . S . defined signalingmotifs found in the intracytoplasmic tail of Pat. No . 7 ,514 ,537 , U . S . Pat. No. 8 , 324 ,353 , WO2010 / a variety of receptors that serve as binding sites for syk / 025177 , U . S . Pat. No . 7 , 446 , 179 , WO2010 / 025177 , zap70 class tyrosine kinases . Examples of ITAM used in the WO2012 / 031744 , WO2012 / 136231A1 , WO2012 / invention can include as non limiting examples those 050374A2 , WO2013074916 , WO /2009 /091826A3 , derived from TCRzeta , FcRgamma , FcRbeta , FcRepsilon , WO2013/ 176915 or WO2013/ 059593 . CD3gamma, CD3delta , CD3epsilon , CD5, CD22 , CD79a , [ 0565 ] The signaling domain of the p -CAR or “ signaling CD79b and CD66d . In a preferred embodiment, the signal protein ” according to the invention is involved in the acti ing transducing domain of a multi -chain CAR according to US 2017 /0296623 A1 Oct . 19, 2017 31 the invention can comprise a CD3zeta signaling domain , or cellular ligand binding -domain comprising VH and VL from the intracytoplasmic domain of the FcRI beta or gamma a monoclonal anti- CD123 antibody , a hinge, a transmem chains . brane domain and a cytoplasmic domain including a CD3 [ 0570 ] In particular embodiment the signal transduction zeta signaling domain and a co - stimulatory domain from domain of the P -CAR of the present invention comprises a 4 - 1BB . co -stimulatory signal molecule . A co -stimulatory molecule [0576 ] As a preferred variant, VH and VL from a mono is a cell surface molecule other than an antigen receptor or clonal anti - CD123 antibody which can be used are derived their ligands that is required for an efficient immune from Klon -43 ( respectively SEQ ID NO :47 - 48 ) . response . “ Co -stimulatory ligand ” refers to a molecule on an [0577 ) As possible options, the following respective short , antigen presenting cell that specifically binds a cognate medium or long hinges from FcyRIIIa , CD8a , IgG1 (SEQ co - stimulatory molecule on a T cell , thereby providing a ID NO : 49 , SEQ ID NO : 50 , SEQ ID NO : 51 ) can be used . signal which , in addition to the primary signal provided by, [ 0578 ] As preferred transmembrane domain , 4 - 1BB or for instance , binding of a TCR / CD3 complex with an MHC CD8a (SEQ ID NO :53 , SEQ ID NO :52 ) can be preferred , molecule loaded with peptide , mediates a T cell response , and more preferably CD8a . including , but not limited to , proliferation activation , differ [ 0579 ] In a preferred embodiment, the P -CAR which is entiation and the like. A “ co -stimulatory molecule ” refers to expressed in the engineered immune cell in combination the cognate binding partner on a T cell that specifically binds with the N -CAR is CD123 specific chimeric antigen recep with a co - stimulatory ligand , thereby mediating a co - stimu tor having one of the polypeptide structure selected from V1, latory response by the cell , such as , but not limited to V3 and V5, as illustrated in FIG . 1, said structure comprising proliferation . Co - stimulatory molecules include, but are not an extra cellular ligand binding -domain comprising VH and limited to an MHC class I molecule, BTLA and Toll ligand VL from a monoclonal anti- CD123 antibody, a hinge , a receptor . transmembrane domain , a cytoplasmic domain including a [0571 ] A co - stimulatory ligand according to the present CD3 zeta signaling domain and a co - stimulatory domain invention can include but is not limited to CD7 , B7 - 1 from 4 - 1BB , said 123 CAR having at least 80 % sequence (CD80 ) , B7 - 2 (CD86 ) , PD -L1 , PD - L2 , 4 - 1BBL , OX40L , identity with either SEQ ID NO . 53, SEQ ID NO . 58 or SEQ inducible costimulatory igand ( ICOS - L ) , intercellular adhe ID NO . 60 . sion molecule ( ICAM , CD3OL , CD40 , CD70 , CD83 , HLA 10580 ) CS1 P -CAR G , MICA , M 1CB , HVEM , lymphotoxin beta receptor, [0581 ] According to another embodiment, the P - CAR 3 / TR6 , ILT3 , ILT4 , an agonist or antibody that binds Toll which is expressed in the engineered immune cell in com ligand receptor and a ligand that specifically binds with bination with the N -CAR is a CS1 specific chimeric antigen B7 -H3 . A co - stimulatory ligand also encompasses, inter alia , receptor (CAR ) having one of the polypeptide structure an antibody that specifically binds with a co - stimulatory selected from V1 to V6 , preferably V1, V3 and V5 as molecule present on a T cell, such as but not limited to , illustrated in FIG . 1 , said structure comprising an extra CD27 , CD28 , 4 - IBB , OX40 , CD30 , CD40 , PD - 1 , ICOS , cellular ligand binding -domain comprising VH and VL from lymphocyte function - associated antigen - 1 (LFA - 1 ) , CD2 , a monoclonal anti -CS1 antibody, a hinge , a transmembrane CD7, LTGHT, NKG2C , B7 -H3 , a ligand that specifically domain and a cytoplasmic domain including a CD3 zeta binds with CD83 . signaling domain and a co -stimulatory domain from 4 - 1BB . [ 0572 ] As preferred and exemplary P -CARs which can be 10582 ] Concerning the VH chain and the VL from a used in combination with the N -CARs such as presented monoclonal anti -CS1 antibody , they can derived from the above , are those with an extracellular -binding domain rec murine scFv Luc63 , Luc90 , Luc34 , LucX1 and LucX2 ognizing the CD19, CD123 , CD38 , CS1, ROR1, CLL - 1 or antibodies (SEQ ID NO : 38 to 47 in WO 2015121454 A1) , CD22 cell surface marker antigen . and optionally humanized from these . [0573 ] Others P -CARs which can be used in combination [ 0583] CD38 P -CAR with a N - CAR according to the present invention are con [ 0584 ] According to another embodiment , the P -CAR templated within the present invention , such as anti - CD28 which is expressed in the engineered immune cell in com CAR , anti -CD30 CAR , anti -CD138 CAR , anti -CD171 bination with the N -CAR is a CD38 specific chimeric CAR , anti- CD19 CAR , anti -CEA CAR ( CEA being the antigen receptor (CAR ) having one of the polypeptide carcinoembryonic antigen ) , anti -ERB B CAR ( ligand of structure selected from V1 to V6 , preferably V1, V3 and V5 HER - 2/ neu ), anti -FAP CAR (Fibroblast activation protein ) , as illustrated in FIG . 1, said structure comprising an extra anti -GD2 CAR , anti -GPC3 CAR (glypican - 3 antigen ), anti cellular ligand binding - domain comprising VH and VL from Lewis - Y CAR (carbohydrate antigen ), anti -NKG2D ligand a monoclonal anti -CD38 antibody, a hinge , a transmembrane CAR , anti -MSLN CAR (mesothelin antigen ), anti- NY domain and a cytoplasmic domain including a CD3 zeta ESO - 1 CAR ( cancer- testis antigen ) , anti -PSCA CAR (Pros signaling domain and a co - stimulatory domain from 4 - 166 . tate stem cell antigen ), anti -GPC3 ( glypican 3 antigen ) 105851 Preferably , the anti -CD38 CAR as P -CAR com CAR , anti -CD20 CAR , anti - HER1 CAR ( EGFR /HER - 1 prises a polypeptide sequence displaying at least 90 % , at oncoantigen ) or anti - CD47 CAR . These P -CARs may have least 95 % , at least 98 % or at least 99 % identity with a a single -chain or a multi - chain architecture . sequence selected from the group consisting of SEQ ID NO . [ 0574 ] CD123 P - CAR 64 -66 (based on 25A10 mAb ) , SEQ ID NO . 67 -69 (based on [0575 ] According to one embodiment, the P -CAR which 28F5 mAb ) , SEQ ID NO . 70 -72 ( based 1665 ) . is expressed in the engineered immune cell in combination [ 0586 ] For these CD38 scCAR and CS1 SCCAR , the with the N -CAR is a CD123 specific chimeric antigen choice of preferred hinge or transmembrane domains receptor (CAR ) having one of the polypeptide structure remains the same than for the CD123 CAR . selected from V1 to V6 , preferably V1, V3 and V5 as [ 0587 ] Although , cells expressing CD38 , as well as many illustrated in FIG . 1, said structure comprising an extra other tumor antigen markers CS1 could be regarded as US 2017 /0296623 A1 Oct . 19, 2017 32 atattractive targets for CARs, the fact that such antigen 70 % , 75 % , 80 % , 85 % when the N - CAR and P -CAR antigen markers are also expressed at the surface ofmost T - cells, has binding domains, both , bind to their respective antigens as hampered significantly the selection of these markers to compared to when the P - CAR antigen binding domain , perform immunotherapy. Thus , according to a preferred alone , binds to its antigen . embodiment, the anti -CD38 positive CAR or the anti -CS1 [0599 ) In some embodiments , the level of activation of the positive CAR is expressed in combination with a N -CAR in immune cell is measured by determining cytokine produc immune cells which are further engineered to inactivate such tion . CD38 or CS1 expressed on the surface of said immune cell. [0600 ] In some embodiments , the level of activation of the This method is described in WO2015 / 121454 . This gene immune cell is measured by monitoring IFNgamma produc inactivation may be performed by the use of specific endo tion by ELISA and /or FACS and / or luminex assay. nuclease such as a TALE - nuclease . 10601 ] In some embodiments, the level of activation of the [0588 ] CLL - 1 P -CAR immune cell is measured by monitoring TNFalpha produc [0589 ] According to another embodiment, the P -CAR tion by ELISA and /or luminex assay . according to the invention which is expressed in the engi [0602 ] In some embodiments , the level of activation of the neered immune cell in combination with the N -CAR is a immune cell is measured by monitoring degranulation , for CLL - 1 specific chimeric antigen receptor (CAR ) having one example by measuring CD107a levels by FACS . of the polypeptide structure selected from V1 to V6 , pref [0603 ] In some embodiments , the level of activation of the erably V1, V3 and V5 as illustrated in FIG . 1 , said structure immune cell is measured by monitoring the ability of the comprising an extra cellular ligand binding - domain com immune cell to kill target cells . prising VH and VL from a monoclonal anti -CLL - 1 antibody, [ 0604 ] In some embodiments , the negative signal of the a hinge , a transmembrane domain and a cytoplasmic domain N -CAR is short- termed and reversible to ensure that the including a CD3 zeta signaling domain and a co -stimulatory immune cells comprising a P -CAR and an N -CAR accord domain from 4 - 1BB . ing to the invention may be activated when it encounters [ 0590 ] Said V2 and Vh from a monoclonal anti -CLL - 1 only P -CAR antigen , despite prior inactivation in a off -tissue antibody are preferably selected from the antibodies referred setting that has both P -CAR and N -CAR antigens . to in the literature as SCO02 - 357 , SCO2- 378 and SCO2- 161 [0605 ] mAb -Specific Epitope Mimotope/ in WO2005 /00894 (Applicant : Crucell Holland BV ) ; M26 , [0606 ] According to another embodiment, the present M31 , G4, M22 , M29, M2, M5, G12 in WO2013 / 169625 invention relates to improved inhibitory chimeric antigen ( Applicant: Cellerant Therapeutics ) ; and 21. 26 , 1075 . 7 in receptors ( ICAR ) , wherein the extracellular binding domain WO2009/ 051974 ( Applicant: Nuvelo Inc ). ( scFv ) has been modified by insertion of at least one [0591 ] The choice of preferred hinge or transmembrane mAb - specific epitope . domains remains the same than for the previous scCARs . [06071 Such insertion is designed to allow both sorting [0592 ] CD22 P -CAR and /or depletion of the immune cells endowed with said [ 0593 ] According to another embodiment, the P -CAR N -CARs . which is expressed in the engineered immune cell in com 10608 ] According to another embodiment, the immune bination with the N -CAR is a CD22 specific chimeric cell has been further engineered to express a P -CAR in antigen receptor (CAR ) having one of the polypeptide which of such at least one mAb - specific epitope is inserted . structure selected from V1 to V6 , preferably V1, V3 and V5 [ 0609] Preferably , twomAb - specific epitopes are inserted . as illustrated in FIG . 1 , said structure comprising an extra [0610 ] Such epitope ( s ) is ( are ) inserted anywhere in the cellular ligand binding - domain comprising VH and VL from extracellular part of N -CAR or P -CAR , either in the N a monoclonal anti - CD22 antibody , a hinge , a transmembrane terminal part , between the VH and VL chains of the scFvs domain and a cytoplasmic domain including a CD3 zeta or between the hinge or linker and the scFvs . Preferably , signaling domain and a co - stimulatory domain from 4 - 1BB . when more than one mAb - specific epitope are used , they are [0594 ] P - CAR Level of Inactivation not in tandem ( side by side ) . [ 0595 ] In some embodiments , the immune cell of the [0611 ] In a preferred embodiment, the epitope introduced invention is activated when the P -CAR antigen binding within the chimeric scFv is the CD20 antigen and the infused domain binds to its antigen . In some embodiments , such mAb which is being used to target it for sorting and /or activation is reduced when the N -CAR antigen binding depletion purpose ( s ) is rixutimab . Such epitope target domain binds to its antigen . sequence has over 80 % identity , preferably over 90 % , and 10596 ] In some embodiments such reduction of activation more preferably over 95 % identity , more preferably 100 % is increased , preferably by at least 5 % , 10 % , 15 % , 20 % or identity with the CD20 antigen of SEQ ID NO . 82 . Such 30 % in an immune cell comprising an N -CAR according to preferred suicide gene system employs a recombinant anti the invention as compared to the same immune cell com genic polypeptide comprising antigenic motif recognized by prising an N -CAR comprising the full intracellular domain the anti - CD20 mAb Rituximab , especially QBen10 , such as of PD - 1 . in the so - called RQR8 polypeptide described in [0597 ] In some embodiments such reduction of activation WO2013153391 . Rituximab , an authorized antibody drug , is increased , preferably by at least 5 % , 10 % , 15 % , 20 % or can then be used for cell depletion when needed . 30 % in an immune cell comprising an N -CAR according to [ 0612 ] According to another embodiment, the epitope is a the invention as compared to the same immune cell com mimotope . As a macromolecule , often a peptide, which prising an N -CAR comprising the full intracellular domain mimics the structure of an epitope , the mimotope has the of CTLA -4 . advantage to be smaller than conventional epitope, and [0598 ] In some embodiments , the activation due to P -CAR therefore may be beneficial for a non - conformational binding to its antigen is reduced by at least 5 % , 10 % , 15 % , sequence and easier to reproduce in a long polypeptide such 20 % , 25 % , 30 % , 35 % , 40 % , 45 % , 50 % , 55 % , 60 % , 65 % , a CAR .Mimotopes are known for several pharmaceutically US 2017 /0296623 A1 Oct . 19 , 2017 33 approved mAb such as two 10 amino acid peptides for [ 0619 ] The present invention relates also to an isolated cetuximab ( Riemer et al ., 2005 ) , or a 24 aa for palivizumab immune cell comprising a P -CAR and an N -CAR such as ( Arbiza et al, 1992 ) . As these mimotopes can be identified by presented above . phage display , it is possible to try several of them in order [0620 ] Said P - CAR may be a single chain CAR or a to obtain a sequence which does not perturb the scFv for the multi -chain P -CAR as defined in WO2014 /039523 . same mAb . Furthermore , their use can enhance a comple [0621 ] According to an embodiment, the isolated immune ment- dependent cytotoxicity (CDC ). cell includes at least a N - CAR which comprises at least: [0613 ] As exemples , mimotopes of CD20 is SEQ ID [ 0622 ] an extracellular binding domain , which is able to NO :73 (CPYSNPSLC ) , mimotopes corresponding to the use bind to an “ off -target ” antigen on a healthy cell and ; of cetuximab of SEQ ID NO : 74 (COFDLSTRRLKC ) SEQ [0623 ] an inhibitory transmembrane polypeptide having ID NO : 75 (CQYNLSSRALKC ) SEQ ID NO : 76 a sequence with more than 80 % , preferably 90 % and (CVWQRWQKSYVC ) , SEQ ID NO : 77 ( CMWDRFSR more preferably 95 % , and even more preferably 100 % WYKC ) ; mimotopes corresponding to the use of palivi identity with a sequence from SEQ ID NO : 1 (human zumab of SEQ ID NO : 78 (NSELLSLINDMPITNDOK KI2L2 ), SEQ ID NO : 2 (human KI2L1 ), SEQ ID NO :3 KLMSNN ) or mimotopes corresponding to the use of ( human FCG2B ), SEQ ID NO : 4 (human KI2L3 ) , SEQ nivolumab of SEQ ID NO : 79 (SFVLNWYRMSPSNQTD ID NO : 5 (human KI3L2 ) , SEQ ID NO :6 (human KLAAFPEDR ), SEQ ID NO : 80 (SGTYLCGAISLAP KI2L4 ), SEQ ID NO :7 (human KI3L1) , SEQ ID NO :8 KAQIKE ) . (human KIZLA ), SEQ ID NO : 9 (human MILR1 ) , SEQ [ 0614 ] The present invention relates also to the immune ID NO : 10 (human LIRB4 ), SEQ ID NO : 11 (human cells expressing said N - CARs , to the methods of in vivo LIRB3) , SEQ ID NO : 12 ( human KI3L3 ), SEQ ID depleting and / or in vitro sorting said CAR - expressing NO : 15 ( human LIRB5 ), SEQ ID NO : 16 (human LIRB2 ), SEQ ID NO : 18 (human FCRL4 ) , SEQ ID immune cells , and is drawn to their therapeutic use . NO : 23 (human FCRL5 ) , SEO ID NO : 24 (human [ 0615 ] Isolated Immune Cell FCRL2 ) , SEQ ID NO : 25 (human FCRL1) , SEQ ID [ 0616 ] Cell according to the present invention refers to a NO :27 (human FCRL3 ) , SEQ ID NO : 28 (human cell of hematopoietic origin functionally involved in the MPZL1) , SEQ ID NO :29 (human PILRA ) or SEQ ID initiation and / or execution of innate and / or adaptative NO : 30 (human PVR ) ; immune response . Cell according to the present invention is [0624 ] and a P -CAR which comprises in the engineered preferably a T - cell obtained from a donor. Said T cell immune cell in combination with the N - CAR ; said P - CAR according to the present invention can be derived from a comprising one transmembrane polypeptide comprising at stem cell . The stem cells can be adult stem cells, embryonic least one extracellular ligand -binding domain able to bind to stem cells , more particularly non -human stem cells , cord CD123 antigen , and one signal - transducing domain , option blood stem cells , progenitor cells , bone marrow stem cells , ally with a co - stimulatory domain . totipotent stem cells or hematopoietic stem cells . In a [0625 ] According to another preferred embodiment, the preferred embodiment, cells are human cells , in particular isolated immune cell includes at least a N -CAR which human stem cells . comprises at least : 106171 Representative human stem cells are CD34 + cells . [ 0626 ] an extracellular binding domain , which is able to Said isolated cell can also be a dendritic cell , killer dendritic bind to an “ off- target ” antigen on a healthy cell; cell, a mast cell , a NK - cell , a B -cell or a T -cell selected from [ 0627 ] an inhibitory transmembrane polypeptide having the group consisting of inflammatory T -lymphocytes , cyto a sequence selected from the group consisting of SEQ toxic T -lymphocytes , regulatory T -lymphocytes or helper ID NO : 1 (human KI2L2 ), SEQ ID NO : 2 (human T - lymphocytes . In another embodiment, said cell can be KI2L1) , SEQ ID NO : 3 (human FCG2B ) , SEQ ID NO : 4 derived from the group consisting of CD4 + T - lymphocytes (human KI2L3 ), SEQ ID NO : 5 (human KI3L2 ), SEQ and CD8 + T - lymphocytes. Prior to expansion and genetic ID NO : 6 ( human KI2L4 ) , SEQ ID NO : 7 (human modification of the cells of the invention , a source of cells KI3L1) , SEQ ID NO : 8 ( human KIZLA ), SEQ ID NO : 9 can be obtained from a subject through a variety of non (human MILR1) , SEQ ID NO : 10 (human LIRB4) , limiting methods. Cells can be obtained from a number of SEQ ID NO : 11 (human LIRB3 ) , SEQ ID NO : 12 (hu non - limiting sources , including peripheral blood mononu man KI3L3 ) , SEQ ID NO : 15 ( human LIRB5 ), SEO ID clear cells, bone marrow , lymph node tissue , cord blood , NO : 16 (human LIRB2) , SEQ ID NO : 18 (human thymus tissue , tissue from a site of infection , ascites, pleural FCRL4 ), SEQ ID NO :23 (human FCRL5) , SEQ ID effusion , spleen tissue, and tumors. In certain embodiments NO : 24 (human FCRL2 ), SEQ ID NO : 25 (human of the present invention , any number of T- cell lines available FCRL1) , SEQ ID NO :27 (human FCRL3 ) , SEQ ID and known to those skilled in the art, may be used . In another NO : 28 (human MPZL1) , SEQ ID NO :29 (human embodiment, said cell is preferably derived from a healthy PILRA ) or SEQ ID NO : 30 (human PVR ); donor. In another embodiment, said cell is part of a mixed [0628 ] and a P - CAR which comprises in the engineered population of cells which present different phenotypic char immune cell in combination with the N -CAR ; said P -CAR acteristics. comprising : [ 0618 ] Preferably , isolation and preparation of stem cells [ 0629 ] one transmembrane polypeptide comprising at does not require the destruction of at least one human least one extracellular ligand -binding domain able to bind to embryo . The immune cells can originate from the patient, in CD123 antigen , and one signal- transducing domain , option view of operating autologous treatments , or from donors in ally with a co - stimulatory domain . view of producing allogeneic cells , which can be used in [0630 ] According to an embodiment, the isolated immune allogeneic treatments. cell includes at least a N -CAR which comprises at least : US 2017 /0296623 A1 Oct . 19 , 2017 34 [0631 ] an extracellular binding domain , comprising a [0638 ] According to a preferred embodiment, the isolated polypeptide sequence sharing more than 80 % , prefer immune cell includes at least a N -CAR which comprises at ably 90 % and more preferably 95 % and even more least : preferably 100 % identity with SEQ ID NO : 81 (CD4 [0639 ] an extracellular binding domain , which is able to antigen ) , SEQ ID NO : 82 (CD20 antigen ), SEQ ID NO : bind to an “ off- target ” antigen on a healthy cell, and 83 (CD22 antigen ), SEQ ID NO : 84 (CD25 antigen ) or [ 0640 ] an inhibitory transmembrane polypeptide having SEQ ID NO : 85 (MUC1 antigen ) ; a polypeptide sequence selected from the group con [ 0632] an inhibitory transmembrane polypeptidehaving sisting of SEQ ID NO : 33 (human TR10D ) , SEQ ID a sequence with more than 80 % , preferably 90 % and NO : 34 (human TR10A ) and SEQ ID NO : 35 (human more preferably 95 % identity and even more preferably TR10B ); 100 % with SEQ ID NO : 1 (human KI2L2 ) , SEQ ID [ 0641 ] and a P -CAR which comprises in the engineered NO : 2 (human KI2L1) , SEQ ID NO : 3 (human immune cell in combination with the N - CAR ; said P - CAR FCG2B ) , SEQ ID NO : 4 ( human KI2L3) , SEQ ID NO : comprising: 5 (human KI3L2 ) , SEQ ID NO : 6 ( human KI2L4 ) , SEQ [0642 ] one transmembrane polypeptide comprising at ID NO : 7 (human KIZL1) , SEQ ID NO : 8 (human least one extracellular ligand -binding domain able to KIZLA ) , SEQ ID NO : 9 (human MILRI) , SEQ ID bind to CD123 antigen , and one signal- transducing NO : 10 (human LIRB4 ) , SEQ ID NO : 11 (human domain , optionally with a co - stimulatory domain . LIRB3) , SEQ ID NO : 12 (human KI3L3 ) , SEQ ID 10643 ] According to another preferred embodiment, the NO : 15 (human LIRB5 ) , SEQ ID NO : 16 (human isolated immune cell includes at least a N - CAR which LIRB2) , SEQ ID NO : 18 (human FCRL4 ), SEQ ID comprises at least : NO : 23 (human FCRL5) , SEQ ID NO :24 (human [ 0644 ] an extracellular binding domain , which is able to FCRL2 ) , SEQ ID NO : 25 (human FCRL1 ) , SEQ ID bind to an " off - target” antigen on a healthy cell and ; NO :27 ( human FCRL3) , SEQ ID NO : 28 (human 106451 an inhibitory transmembrane polypeptide having MPZL1 ) , SEQ ID NO :29 (human PILRA ) or SEQ ID a sequence of SEQ ID NO : 1 (human KI2L2 ), SEQ ID NO : 30 ( human PVR ) ; NO : 2 (human KI2L1) , SEQ ID NO : 3 (human [0633 ] and a P - CAR which comprises in the engineered FCG2B ) , SEQ ID NO : 4 (human KI2L3 ) , SEO ID NO : immune cell in combination with the N -CAR ; said P -CAR 5 (human KI3L2 ) , SEQ ID NO : 6 ( human KI2L4 ) , SEQ comprising one transmembrane polypeptide comprising at ID NO : 7 (human KI3L1) , SEQ ID NO : 8 (human least one extracellular ligand -binding domain able to bind to KIZLA ), SEQ ID NO : 9 (human MILR1) , SEQ ID CD123 antigen , and one signal - transducing domain , option NO : 10 (human LIRB4 ) , SEQ ID NO : 11 (human ally with a co - stimulatory domain . LIRB3 ) , SEQ ID NO : 12 (human KI3L3 ) , SEQ ID 0634 ) According to another preferred embodiment, the NO : 15 ( human LIRB5 ) , SEQ ID NO : 16 (human LIRB2 ) , SEQ ID NO : 18 ( human FCRL4 ) , SEQ ID isolated immune cell includes at least a N - CAR which NO : 23 (human FCRL5 ) , SEQ ID NO : 24 (human comprises at least: FCRL2 ) , SEQ ID NO : 25 (human FCRL1) , SEQ ID [ 0635 ] an extracellular binding domain , comprising a NO :27 (human FCRL3 ), SEQ ID NO : 28 (human polypeptide sequence sharing more than 80 % , prefer MPZL1) , SEQ ID NO :29 (human PILRA ) or SEQ ID ably 90 % and more preferably 95 % identity and even NO :30 (human PVR ); more preferably 100 % with SEQ ID NO : 81 ( CD4 [ 0646 ] and a P - CAR which comprises in the engineered antigen ) , SEQ ID NO : 82 ( CD20 antigen ) , SEQ ID NO : immune cell in combination with the N -CAR ; said P -CAR 83 (CD22 antigen ), SEQ ID NO : 84 (CD25 antigen ) or comprising one transmembrane polypeptide comprising at SEQ ID NO : 85 (MUC1 antigen ); least one extracellular ligand -binding domain able to bind to [0636 ] an inhibitory transmembrane polypeptide having CD123 antigen , and one signal- transducing domain , option a sequence of SEQ ID NO : 1 (human KI2L2 ) , SEQ ID ally with a co - stimulatory domain . NO : 2 ( human KI2L1) , SEQ ID NO : 3 (human [0647 ] According to a preferred embodiment, the isolated FCG2B ) , SEQ ID NO : 4 ( human KI2L3) , SEQ ID NO : immune cell includes at least a N -CAR which comprises at 5 ( human KI3L2 ), SEQ ID NO :6 (human KI2L4 ), SEQ least: ID NO : 7 (human KI3L1 ), SEQ ID NO : 8 (human 10648 ] an extracellular binding domain , comprising a KIZLA ), SEQ ID NO : 9 (human MILR1) , SEQ ID polypeptide sequence sharing more than 80 % , preferably NO : 10 (human LIRB4 ) , SEQ ID NO : 11 (human 90 % and more preferably 95 % identity and even more LIRB3) , SEQ ID NO : 12 (human KI3L3 ) , SEQ ID preferably 100 % with SEQ ID NO : 81 (CD4 antigen ) , SEQ NO : 15 (human LIRB5 ) , SEQ ID NO : 16 (human ID NO : 82 (CD20 antigen ) , SEQ ID NO : 83 (CD22 antigen ) , LIRB2) , SEQ ID NO : 18 (human FCRL4 ) , SEQ ID SEQ ID NO : 84 (CD25 antigen ) or SEQ ID NO : 85 (MUC ) NO :23 (human FCRL5 ) , SEQ ID NO : 24 (human antigen ) ; FCRL2 ) , SEO ID NO : 25 (human FCRL1) , SEO ID [0649 ] and ; NO : 27 (human FCRL3 ) , SEQ ID NO :28 (human [0650 ] an inhibitory transmembrane polypeptide having MPZL1) , SEQ ID NO : 29 (human PILRA ) or SEQ ID a sequence with more than 80 % , preferably 90 % and NO : 30 ( human PVR ) ; more preferably 95 % identity and even more preferably [ 0637 ] and a P -CAR which comprises in the engineered 100 % identity with SEO ID NO : 1 (human KI2L2 ) , immune cell in combination with the N -CAR ; said P -CAR SEQ ID NO : 2 (human KI2L1) , SEQ ID NO : 3 (human comprising one transmembrane polypeptide comprising at FCG2B ), SEQ ID NO : 4 (human KI2L3) , SEQ ID NO : least one extracellular ligand -binding domain able to bind to 5 (human KI3L2 ), SEQ ID NO : 6 (human KI2L4 ) , SEO CD123 antigen , and one signal - transducing domain , option ID NO : 7 (human KI3L1) , SEQ ID NO : 8 (human ally with a co - stimulatory domain . KIZLA ), SEQ ID NO :9 (human MILR1) , SEQ ID US 2017 /0296623 A1 Oct . 19 , 2017 35 NO : 10 (human LIRB4 ), SEQ ID NO : 11 (human [ 0660 ] According to a more preferred embodiment, the LIRB3 ) , SEQ ID NO : 12 (human KI3L3 ) , SEQ ID isolated immune cell includes at least a N -CAR which NO : 15 (human LIRB5 ) , SEQ ID NO : 16 (human comprises at least: LIRB2 ), SEQ ID NO : 18 (human FCRL4 ) , SEQ ID [ 0661] an extracellular binding domain , comprising a NO : 23 (human FCRL5) , SEQ ID NO : 24 (human polypeptide sequence sharing more than 80 % , prefer FCRL2 ) , SEQ ID NO : 25 (human FCRL1 ) , SEQ ID ably 90 % and more preferably 95 % identity and even NO : 27 ( human FCRL3) , SEQ ID NO : 28 ( human more preferably 100 % with SEQ ID NO : 81 (CD4 MPZL1) , SEQ ID NO : 29 (human PILRA ) or SEQ ID antigen ) , SEQ ID NO : 82 (CD20 antigen ) , SEQ ID NO : NO : 30 (human PVR ) ; 83 (CD22 antigen ) , SEQ ID NO : 84 (CD25 antigen ) or [ 0651 ] and a CD123 specific chimeric antigen receptor SEQ ID NO : 85 (MUC1 antigen ); (CAR ) (as P -CAR ) which comprises in the engineered [0662 ] and ; immune cell in combination with the N -CAR ; said [0663 ] an inhibitory transmembrane polypeptide having CD123 specific chimeric antigen receptor ( CAR ) hav a polypeptide sequence of more than 80 % , preferably ing one of the polypeptide structure selected from V1 90 % and more preferably 95 % identity and even more to V6 , preferably V1, V3 and V5 as illustrated in FIG . preferably 100 % identity with SEQ ID NO : 33 (human 1 , said structure comprising an extra cellular ligand TR10D ), SEQ ID NO : 34 (human TR10A ) and SEQ ID binding - domain comprising VH and VL from a mono NO : 35 (human TR10B ) ; clonal anti -CD123 antibody , a hinge, a transmembrane [ 0664 ] and a CD123 specific chimeric antigen receptor domain and a cytoplasmic domain including a CD3 (CAR ) (as P -CAR ) which comprises in the engineered zeta signaling domain and a co -stimulatory domain immune cell in combination with the N - CAR ; said from 4 - 1BB . CD123 specific chimeric antigen receptor (CAR ) hav [ 0652 ] According to a preferred embodiment, the isolated ing one of the polypeptide structure selected from V1 immune cell includes at least a N - CAR which comprises at to V6 , preferably V1 , V3 and V5 as illustrated in FIG . 1 , said structure comprising an extra cellular ligand least : binding - domain comprising VH and VL from a mono [ 0653 ] an extracellular binding domain , comprising a clonal anti- CD123 antibody , a hinge , a transmembrane polypeptide sequence sharing more than 80 % , prefer domain and a cytoplasmic domain including a CD3 ably 90 % and more preferably 95 % identity and even zeta signaling domain and a co -stimulatory domain more preferably 100 % with SEQ ID NO : 81 (CD4 from 4 - 1BB . antigen ) , SEQ ID NO : 82 ( CD20 antigen ) , SEQ ID NO : [ 0665 ] Preferably , the above anti- CD123 CARs ( P -CARs ) 83 (CD22 antigen ) , SEQ ID NO : 84 (CD25 antigen ) or having one of the polypeptide structure selected from V1, SEQ ID NO : 85 (MUC1 antigen ) ; V3 and V5 , as illustrated in FIG . 1 , said structure comprising [0654 ] an inhibitory transmembrane polypeptidehaving an extra cellular ligand binding -domain comprising VH and a polypeptide sequence of more than 80 % , preferably VL from a monoclonal anti- CD123 antibody, a hinge , a 90 % and more preferably 95 % identity with SEQ ID transmembrane domain , a cytoplasmic domain including a NO : 33 (human TR10D ), SEQ ID NO : 34 (human CD3 zeta signaling domain and a co - stimulatory domain TR10A ) and SEQ ID NO : 35 (human TR10B ); from 4 - 1BB , said 123 CAR having at least 80 % sequence [0655 ) and a P -CAR which comprises in the engineered identity with either SEQ ID NO . 56 , SEQ ID NO . 58 or SEQ immune cell in combination with the N -CAR ; said P -CAR ID NO . 60 . comprising one transmembrane polypeptide comprising at [ 0666 ] According to an embodiment, the isolated immune least one extracellular ligand -binding domain able to bind to cell includes at least a N -CAR which comprises at least: CD123 antigen , and one signal- transducing domain , option [ 0667 ] an extracellular binding domain , comprising a ally with a co - stimulatory domain . polypeptide sequence sharing more than 80 % , prefer ably 90 % and more preferably 95 % identity and even [0656 ] According to a preferred embodiment, the isolated more preferably 100 % identity with SEQ ID NO : 86 immune cell includes at least a N -CAR which comprises at (CD56 antigen ), SEQ ID NO : 87 (CD205 antigen ), least : SEQ ID NO : 88 ( CD83 antigen ) , SEQ ID NO : 89 [0657 ] an extracellular binding domain , comprising a (CD206 antigen ) , SEQ ID NO : 90 (CD200 antigen ), or polypeptide sequence sharing more than 80 % , prefer SEQ ID NO : 91 (CD36 antigen ); ably 90 % and more preferably 95 % identity and even [0668 ] and ; more preferably 100 % with SEQ ID NO : 81 ( CD4 [ 0669 ] an inhibitory transmembrane polypeptide having antigen ), SEQ ID NO : 82 (CD20 antigen ), SEQ ID NO : a sequence with more than 80 % , preferably 90 % and 83 (CD22 antigen ), SEQ ID NO : 84 ( CD25 antigen ) or more preferably 95 % identity and even more preferably SEQ ID NO : 85 (MUC1 antigen ) ; 100 % identity with SEQ ID NO : 1 ( human KI2L2 ) , [ 0658 ] an inhibitory transmembrane polypeptide having SEQ ID NO : 2 (human KI2L1) , SEQ ID NO : 3 (human a polypeptide sequence of SEQ ID NO : 33 (human FCG2B ) , SEQ ID NO : 4 (human KI2L3 ) , SEQ ID NO : TR10D ) , SEQ ID NO : 34 (human TR10A ) and SEQ ID 5 ( human KI3L2 ) , SEO ID NO : 6 (human KI2L4 ) , SEO NO : 35 (human TR10B ) . ID NO : 7 (human KI3L1) , SEQ ID NO : 8 (human [ 0659 ] and a P -CAR which comprises in the engineered KIZLA ) , SEO ID NO : 9 (human MILRI ) , SEO ID immune cell in combination with the N -CAR ; said P -CAR NO : 10 (human LIRB4 ) , SEQ ID NO : 11 (human comprising one transmembrane polypeptide comprising at LIRB3) , SEQ ID NO : 12 (human KI3L3) , SEQ ID least one extracellular ligand -binding domain able to bind to NO : 15 (human LIRB5 ), SEQ ID NO : 16 (human CD123 antigen , and one signal - transducing domain , option LIRB2 ), SEQ ID NO : 18 (human FCRL4 ) , SEQ ID ally with a co -stimulatory domain . NO : 23 (human FCRL5 ) , SEQ ID NO : 24 (human US 2017 /0296623 A1 Oct . 19 , 2017 36 . FCRL2 ) , SEQ ID NO : 25 (human FCRL1) , SEQ ID NO :28 (human MPZL1) , SEQ ID NO :29 (human NO : 27 (human FCRL3) , SEQ ID NO : 28 (human PILRA ) or SEQ ID NO :30 (human PVR ) ; MPZL1) , SEQ ID NO :29 (human PILRA ) or SEQ ID 10680 ] and a CD38 specific chimeric antigen receptor NO : 30 (human PVR ); (CAR ) (as P -CAR ) which comprises in the engineered [0670 ] and a CD123 specific chimeric antigen receptor immune cell in combination with the N -CAR ; said CD38 (CAR ) ( as P - CAR ) which comprises in the engineered specific chimeric antigen receptor (CAR ) containing at least immune cell in combination with the N -CAR ; said one transmembrane polypeptide which includes at least one CD123 specific chimeric antigen receptor (CAR ) hav extra -binding domain recognizing specifically the CD38 ing one of the polypeptide structure selected from V1 antigen , and an intracellular signaling domain , optionally to V6 , preferably V1, V3 and V5 as illustrated in FIG . with co - stimulatory domain . 1 , said structure comprising an extra cellular ligand [0681 ] According to a preferred embodiment, the isolated binding - domain comprising VH and VL from a mono immune cell includes at least a N -CAR which comprises at clonal anti - CD123 antibody, a hinge , a transmembrane least : domain and a cytoplasmic domain including a CD3 [0682 ] an extracellular binding domain , comprising a zeta signaling domain and a co - stimulatory domain polypeptide sequence sharing more than 80 % , preferably from 4 - 1BB . 90 % and more preferably 95 % identity and even more [0671 ] According to an embodiment, the isolated immune preferably 100 % identity with SEQ ID NO : 86 (CD56 cell includes at least a N -CAR which comprises at least: antigen ), SEQ ID NO : 87 (CD205 antigen ) , SEQ ID NO : 88 10672 ] an extracellular binding domain , which is able to (CD83 antigen ) , SEQ ID NO : 89 (CD206 antigen ) , SEQ ID bind to an “ off - target” antigen on a healthy cell ; NO : 90 (CD200 antigen ), or SEQ ID NO : 91 (CD36 [ 0673] and ; antigen ) ; and ; [0674 ] an inhibitory transmembrane polypeptide having [0683 ] an inhibitory transmembrane polypeptide having a sequence with more than 80 % , preferably 90 % and a sequence with more than 80 % , preferably 90 % and more preferably 95 % identity and even more preferably more preferably 95 % identity and even more preferably 100 % identity with SEQ ID NO : 1 (human KI2L2) , 100 % identity with SEQ ID NO : 1 (human KI2L2 ) , SEQ ID NO : 2 (human KI2L1) , SEQ ID NO : 3 (human FCG2B ) , SEO ID NO : 4 (human KI2L3) , SEQ ID NO : SEQ ID NO : 2 (human KI2L1) , SEQ ID NO : 3 (human 5 (human KI3L2 ) , SEQ ID NO :6 (human KI2L4 ) , SEQ FCG2B ), SEQ ID NO :4 (human KI2L3 ) , SEQ ID NO : ID NO : 7 (human KIZL1) , SEQ ID NO : 8 (human 5 ( human KI3L2 ) , SEQ ID NO : 6 ( human KI2L4 ) , SEQ KIZLA ), SEQ ID NO : 9 (human MILR1) , SEQ ID ID NO : 7 ( human KI3L1) , SEQ ID NO : 8 (human NO : 10 (human LIRB4 ), SEQ ID NO : 11 (human KI2LA ), SEQ ID NO : 9 (human MILR1) , SEQ ID LIRB3 ) , SEO ID NO : 12 (human KI3L3 ) , SEO ID NO : 10 (human LIRB4 ), SEQ ID NO : 11 (human NO : 15 (human LIRB5 ) , SEQ ID NO : 16 (human LIRB3 ) , SEQ ID NO : 12 ( human KI3L3) , SEQ ID LIRB2 ) , SEO ID NO : 18 (human FCRL4 ) , SEQ ID NO : 15 (human LIRB5 ), SEQ ID NO : 16 (human NO : 23 (human FCRL5 ) , SEQ ID NO : 24 (human LIRB2 ) , SEQ ID NO : 18 ( human FCRL4 ) , SEQ ID FCRL2) , SEQ ID NO : 25 (human FCRL1) , SEQ ID NO : 23 (human FCRL5 ) , SEQ ID NO : 24 (human NO : 27 (human FCRL3 ), SEQ ID NO : 28 (human FCRL2 ) , SEQ ID NO : 25 (human FCRL1) , SEQ ID MPZL1) , SEQ ID NO :29 (human PILRA ) or SEQ ID NO : 27 ( human FCRL3 ), SEQ ID NO : 28 (human NO : 30 ( human PVR ) ; MPZL1) , SEQ ID NO :29 (human PILRA ) or SEQ ID [ 0675 ] and a CD38 specific chimeric antigen receptor NO : 30 (human PVR ) ; ( CAR ) (as P - CAR ) which comprises in the engineered 10684 ] and a CD38 specific chimeric antigen receptor immune cell in combination with the N -CAR ; said CD38 ( CAR ) ( as P - CAR ) which comprises in the engineered specific chimeric antigen receptor (CAR ) containing at least immune cell in combination with the N -CAR ; said CD38 one transmembrane polypeptide which includes at least one specific chimeric antigen receptor (CAR ) having one of the extra -binding domain recognizing specifically the CD38 polypeptide structure selected from V1 to V6 , preferably V1 , antigen , and an intracellular signaling domain , optionally V3 and V5 as illustrated in FIG . 1 , said structure comprising with co - stimulatory domain . an extra cellular ligand binding -domain comprising VH and 10676 ). According to an embodiment, the isolated immune VL from a monoclonal anti -CD38 antibody, a hinge , a cell includes at least a N -CAR which comprises at least: transmembrane domain and a cytoplasmic domain including [0677 ] an extracellular binding domain , which is able to a CD3 zeta signaling domain and a co - stimulatory domain bind to an “ off- target " antigen on a healthy cell; from 4 - 1BB . [ 0678 ] and ; [0685 ] According to a preferred embodiment, the isolated [ 0679 ] an inhibitory transmembrane polypeptide of immune cell includes at least a N -CAR which comprises at SEQ ID NO : 1 (human KI2L2 ), SEQ ID NO : 2 (human least : KI2L1) , SEQ ID NO : 3 (human FCG2B ) , SEQ ID NO : 4 [ 0686 ] an extracellular binding domain , comprising a (human KI2L3 ) , SEQ ID NO : 5 (human KI3L2) , SEQ polypeptide sequence sharing more than 80 % , prefer ID NO :6 (human KI2L4 ) , SEQ ID NO : 7 (human ably 90 % and more preferably 95 % identity and even KI3L1) , SEO ID NO : 8 ( human KIZLA ) , SEO ID NO : 9 more preferably 100 % identity with SEQ ID NO : 86 (human MILR1) , SEQ ID NO : 10 (human LIRB4) , (CD56 antigen ), SEQ ID NO : 87 (CD205 antigen ) , SEQ ID NO : 11 (human LIRB3 ), SEQ ID NO : 12 (hu SEQ ID NO : 88 (CD83 antigen ) , SEQ ID NO : 89 man KI3L3 ) , SEQ ID NO : 15 (human LIRB5 ), SEO ID ( CD206 antigen ), SEQ ID NO : 90 (CD200 antigen ) , or NO : 16 (human LIRB2) , SEQ ID NO : 18 (human SEQ ID NO : 91 (CD36 antigen ) ; FCRL4 ) , SEQ ID NO : 23 (human FCRL5 ) , SEQ ID [0687 ] and ; NO : 24 (human FCRL2) , SEQ ID NO : 25 (human [ 0688 ] an inhibitory transmembrane polypeptide having FCRL1) , SEQ ID NO : 27 (human FCRL3 ) , SEQ ID a sequence of SEQ ID NO : 1 (human KI2L2 ), SEQ ID US 2017 /0296623 A1 Oct . 19 , 2017 37 NO : 2 ( human KI2L1) , SEQ ID NO : 3 (human [0697 ] According to an embodiment, the isolated immune FCG2B ) , SEQ ID NO : 4 ( human KI2L3) , SEQ ID NO : cell includes at least a N -CAR which comprises at least : 5 (human KI3L2 ) , SEQ ID NO : 6 ( human KI2L4 ) , SEQ [0698 ] an extracellular binding domain , which is able to ID NO : 7 (human KI3L1) , SEQ ID NO : 8 (human bind to an “ off -target ” antigen on a healthy cell ; KIZLA ), SEQ ID NO : 9 (human MILR1) , SEQ ID 10699 ] and ; NO : 10 (human LIRB4 ) , SEQ ID NO : 11 (human [0700 ] an inhibitory transmembrane polypeptide having LIRB3 ) , SEO ID NO : 12 ( human KI3L3 ) , SEO ID a sequence with more than 80 % , preferably 90 % and NO : 15 (human LIRB5 ) , SEQ ID NO : 16 (human more preferably 95 % identity and even more preferably LIRB2 ) , SEQ ID NO : 18 (human FCRL4 ) , SEQ ID 100 % identity with SEQ ID NO : 1 (human KI2L2 ) , NO : 23 (human FCRL5 ), SEQ ID NO : 24 (human SEQ ID NO : 2 (human KI2L1) , SEQ ID NO : 3 (human FCRL2) , SEQ ID NO : 25 (human FCRL1) , SEQ ID FCG2B ), SEQ ID NO :4 (human KI2L3 ) , SEQ ID NO : NO : 27 (human FCRL3 ) , SEQ ID NO : 28 (human 5 (human KI3L2 ) , SEQ ID NO : 6 ( human KI2L4 ) , SEQ MPZL1) , SEQ ID NO :29 (human PILRA ) or SEQ ID ID NO : 7 (human KI3L1 ) , SEQ ID NO : 8 (human NO : 30 (human PVR ) ; KIZLA ), SEQ ID NO : 9 (human MILR1) , SEQ ID [ 0689] and a CD38 specific chimeric antigen receptor NO : 10 (human LIRB4 ) , SEQ ID NO : 11 (human (CAR ) ( as P - CAR ) which comprises in the engineered LIRB3 ) , SEQ ID NO : 12 ( human KI3L3) , SEQ ID immune cell in combination with the N -CAR ; said CD38 NO : 15 (human LIRB5 ) , SEQ ID NO : 16 (human specific chimeric antigen receptor (CAR ) having one of the LIRB2 ) , SEQ ID NO : 18 (human FCRL ) , SEO ID polypeptide structure selected from V1 to V6 , preferably V1, NO : 23 ( human FCRL5 ) , SEQ ID NO : 24 (human V3 and V5 as illustrated in FIG . 1 , said structure comprising an extra cellular ligand binding - domain comprising VH and FCRL2 ) , SEQ ID NO : 25 (human FCRL1) , SEQ ID VL from a monoclonal anti -CD38 antibody , a hinge , a NO : 27 (human FCRL3 ), SEQ ID NO : 28 ( human transmembrane domain and a cytoplasmic domain including MPZL1) , SEQ ID NO :29 (human PILRA ) or SEQ ID a CD3 zeta signaling domain and a co - stimulatory domain NO : 30 ( human PVR ) ; from 4 - 1BB . [0701 ] and a CS1 specific chimeric antigen receptor [ 0690 ) According to a preferred embodiment, the isolated (CAR ) (as P -CAR ) which comprises in the engineered immune cell includes at least a N -CAR which comprises at immune cell in combination with the N -CAR ; said CS1 least: specific chimeric antigen receptor (CAR ) containing at 10691 ] an extracellular binding domain , comprising a least one transmembrane polypeptide which includes at polypeptide sequence sharing more than 80 % , prefer least one extra - binding domain recognizing specifically ably 90 % and more preferably 95 % identity and even the CS1 antigen , and an intracellular signaling domain , more preferably 100 % identity with SEQ ID NO : 86 optionally with co -stimulatory domain . (CD56 antigen ) , SEQ ID NO : 87 (CD205 antigen ), [0702 ] According to an embodiment, the isolated immune SEQ ID NO : 88 (CD83 antigen ) , SEQ ID NO : 89 cell includes at least a N - CAR which comprises at least : (CD206 antigen ) , SEQ ID NO : 90 ( CD200 antigen ) , or 10703 ) an extracellular binding domain , which is able to SEQ ID NO : 91 (CD36 antigen ); bind to an “ off -target ” antigen on a healthy cell ; [0692 ] and ; [ 0693 ] an inhibitory transmembrane polypeptide having [ 0704 ] and ; a polypeptide sequence ofmore than 80 % , preferably [ 0705 ] an inhibitory transmembrane polypeptide of 90 % and more preferably 95 % identity and even more SEQ ID NO : 1 (human KI2L2 ) , SEQ ID NO : 2 (human preferably 100 % identity with SEQ ID NO : 33 (human KI2L1 ) , SEO ID NO : 3 (human FCG2B ) , SEO ID NO : 4 TR10D ) , SEQ ID NO : 34 (human TR10A ) and SEQ ID (human KI2L3 ) , SEQ ID NO : 5 (human KI3L2) , SEQ NO : 35 (human TR10B ) ; ID NO : 6 ( human KI2L4 ) , SEQ ID NO : 7 (human [ 06941 and a CD38 specific chimeric antigen receptor KI3L1) , SEQ ID NO : 8 (human KI2LA ), SEQ ID NO : 9 (CAR ) ( as P -CAR ) which comprises in the engineered (human MILR1) , SEQ ID NO : 10 (human LIRB4) , immune cell in combination with the N -CAR ; said CD38 SEQ ID NO : 11 (human LIRB3) , SEQ ID NO : 12 (hu specific chimeric antigen receptor (CAR ) having one of the man KI3L3 ) , SEQ ID NO : 15 (human LIRB5 ), SEO ID polypeptide structure selected from V1 to V6 , preferably V1, NO : 16 (human LIRB2) , SEQ ID NO : 18 (human V3 and V5 as illustrated in FIG . 1 , said structure comprising FCRL4 ) , SEQ ID NO :23 (human FCRL5) , SEQ ID an extra cellular ligand binding - domain comprising VH and NO : 24 (human FCRL2 ), SEQ ID NO : 25 (human VL from a monoclonal anti -CD38 antibody , a hinge , a FCRL1 ) , SEO ID NO : 27 ( human FCRL3 ) , SEO ID transmembrane domain and a cytoplasmic domain including NO : 28 ( human MPZL1) , SEQ ID NO : 29 (human a CD3 zeta signaling domain and a co - stimulatory domain PILRA ) or SEQ ID NO : 30 (human PVR ) ; from 4 - 1BB . [ 0706 ] and a CS1 specific chimeric antigen receptor [0695 ] Preferably , the anti- CD38 CAR as P -CAR com (CAR ) ( as P -CAR ) which comprises in the engineered prises a polypeptide sequence displaying at least 90 % , at immune cell in combination with the N - CAR ; said CS1 least 95 % , at least 98 % or at least 99 % identity to one specific chimeric antigen receptor (CAR ) containing at least selected from SEQ ID NO . 64 -66 ( based on 25A10 mAb ) , SEQ ID NO . 67 -69 (based on 28F5 mAb ) or SEQ ID NO . one transmembrane polypeptide which includes at least one 70 -72 (based on 16B5 mAb ). extra -binding domain recognizing specifically the CS1 anti [ 0696 ] According to a more preferred embodiment, the gen , and an intracellular signaling domain , optionally with anti- CD38 specific chimeric antigen receptor ( anti- CD38 co - stimulatory domain . CAR ) of the invention comprises a polypeptide sequence [0707 ] According to an embodiment , the isolated immune displaying at least 90 % , at least 95 % , at least 98 % or at least cell includes at least a N - CAR which comprises at least: 99 % identity to one selected from SEQ ID NO . 64 -66 (based [ 0708 ] an extracellular binding domain , comprising a on 25A10 mAb ) or SEQ ID NO . 67 -69 (based on 28F5 polypeptide sequence sharing more than 80 % , prefer mAb ) . ably 90 % and more preferably 95 % and even more US 2017 /0296623 A1 Oct . 19 , 2017 38 preferably 100 % identity identity with SEQ ID NO : 92 VL from a monoclonal anti -CS1 antibody , a hinge, a trans (troponin C ) , SEQ ID NO : 93 ( beta - 1 integrin ), SEQ ID membrane domain and a cytoplasmic domain including a NO : 94 ( CCKBR antigen ), SEQ ID NO : 95 (GALR1 CD3 zeta signaling domain and a co - stimulatory domain antigen ) or SEQ ID NO : 96 (CUBN antigen ); from 4 - 1BB . [0709 ] an inhibitory transmembrane polypeptide having [0715 ] According to an embodiment, the isolated immune a sequence with more than 80 % , preferably 90 % and cell includes at least a N -CAR which comprises at least : more preferably 95 % identity and even more preferably [0716 ] an extracellular binding domain , which is able to 100 % identity with SEQ ID NO : 1 (human KI2L2) , bind to an “ off - target ” antigen on a healthy cell ; SEQ ID NO : 2 (human KI2L1) , SEQ ID NO : 3 (human [0717 ] and ; FCG2B ) , SEQ ID NO : 4 ( human KI2L3) , SEQ ID NO : [ 0718 ] an inhibitory transmembrane polypeptide having 5 (human KI3L2) , SEQ ID NO : 6 (human KI2L4 ) , SEQ a polypeptide sequence of more than 80 % , preferably ID NO : 7 (human KI3L1) , SEQ ID NO : 8 (human 90 % and more preferably 95 % identity and even more KIZLA ) , SEQ ID NO : 9 (human MILR1) , SEQ ID preferably 100 % identity with SEQ ID NO : 33 (human NO : 10 (human LIRB4 ) , SEQ ID NO : 11 (human TR10D ) , SEQ ID NO : 34 (human TR10A ) and SEQ ID LIRB3 ) , SEQ ID NO : 12 (human KI3L3 ) , SEQ ID NO : 35 (human TR10B ) ; NO : 15 (human LIRB5 ) , SEQ ID NO : 16 (human 07191 and a CS1 specific chimeric antigen receptor LIRB2) , SEQ ID NO : 18 (human FCRL4 ) , SEQ ID (CAR ) ( as P -CAR ) which comprises in the engineered NO : 23 (human FCRL5) , SEQ ID NO : 24 (human immune cell in combination with the N -CAR ; said CS1 FCRL2) , SEQ ID NO : 25 ( human FCRL1) , SEQ ID specific chimeric antigen receptor (CAR ) containing at NO : 27 (human FCRL3 ), SEQ ID NO : 28 (human least one transmembrane polypeptide which includes at MPZL1) , SEQ ID NO : 29 (human PILRA ) or SEQ ID least one extra -binding domain recognizing specifically NO :30 ( human PVR ) ; the CS1 antigen , and an intracellular signaling domain , [0710 ] and a CS1 specific chimeric antigen receptor optionally with co - stimulatory domain . (CAR ) ( as P -CAR ) which comprises in the engineered 0720 ) According to an embodiment , the isolated immune immune cell in combination with the N -CAR ; said CS1 cell includes at least a N -CAR which comprises at least: specific chimeric antigen receptor (CAR ) having one of the [0721 ] an extracellular binding domain , which is able to polypeptide structure selected from V1 to V6 , preferably V1 , bind to an “ off -target " antigen on a healthy cell ; V3 and V5 as illustrated in FIG . 1 , said structure comprising [0722 ] and ; an extra cellular ligand binding - domain comprising VH and [ 0723 ] an inhibitory transmembrane polypeptide having VL from a monoclonal anti- CS1 antibody , a hinge, a trans a polypeptide sequence of SEQ ID NO : 33 (human membrane domain and a cytoplasmic domain including a TR10D ), SEQ ID NO : 34 ( human TR10A ) and SEQ ID CD3 zeta signaling domain and a co - stimulatory domain NO : 35 (human TR10B ) ; from 4 - 1BB . [ 0724 ] and a CS1 specific chimeric antigen receptor 0711 ] According to an embodiment, the isolated immune (CAR ) ( as P -CAR ) which comprises in the engineered cell includes at least a N -CAR which comprises at least : immune cell in combination with the N -CAR ; said CS1 10712 ] an extracellular binding domain , comprising a specific chimeric antigen receptor (CAR ) containing at polypeptide sequence sharing more than 80 % , prefer least one transmembrane polypeptide which includes at ably 90 % and more preferably 95 % and even more least one extra -binding domain recognizing specifically preferably 100 % identity identity with SEQ ID NO : 92 the CS1 antigen , and an intracellular signaling domain , (troponin C ) , SEQ ID NO : 93 (beta - 1 integrin ) , SEQ ID optionally with co - stimulatory domain . NO : 94 ( CCKBR antigen ) , SEQ ID NO : 95 (GALR1 [0725 ] According to a preferred embodiment, the isolated antigen ) or SEQ ID NO : 96 ( CUBN antigen ) ; immune cell includes at least a N - CAR which comprises at [0713 ] an inhibitory transmembrane polypeptide of least : SEQ ID NO : 1 (human KI2L2 ) , SEQ ID NO : 2 (human [0726 ] an extracellular binding domain , comprising a KI2L1 ), SEQ ID NO :3 (human FCG2B ), SEQ ID NO :4 polypeptide sequence sharing more than 80 % , prefer (human KI2L3 ) , SEQ ID NO : 5 (human KI3L2) , SEQ ably 90 % and more preferably 95 % and even more ID NO :6 (human KI2L4 ) , SEQ ID NO : 7 (human preferably 100 % identity identity with SEQ ID NO : 92 KI3L1 ) , SEQ ID NO : 8 (human KIZLA ), SEQ ID NO : 9 ( troponin C ) , SEQ ID NO : 93 ( beta - 1 integrin ) , SEQ ID (human MILR1) , SEQ ID NO : 10 ( human LIRB4 ), NO : 94 (CCKBR antigen ), SEQ ID NO : 95 (GALR1 SEQ ID NO : 11 (human LIRB3 ) , SEQ ID NO : 12 (hu antigen ) or SEQ ID NO : 96 (CUBN antigen ) ; and ; man KI3L3) , SEQ ID NO : 15 (human LIRB5) , SEQ ID [ 0727 ] an inhibitory transmembrane polypeptide having NO : 16 (human LIRB2 ) , SEQ ID NO : 18 (human a polypeptide sequence of more than 80 % , preferably FCRL4 ) , SEQ ID NO : 23 (human FCRL5 ) , SEQ ID 90 % and more preferably 95 % identity and even more NO : 24 (human FCRL2 ), SEQ ID NO : 25 (human preferably 100 % identity with SEQ ID NO : 33 (human FCRL1) , SEQ ID NO :27 (human FCRL3) , SEQ ID TR10D ), SEQ ID NO : 34 ( human TR10A ) and SEQ ID NO : 28 (human MPZL1) , SEQ ID NO :29 (human NO : 35 (human TR10B ) ; PILRA ) or SEQ ID NO :30 (human PVR ); [0728 ] and a CS1 specific chimeric antigen receptor [ 0714 ] and a CS1 specific chimeric antigen receptor (CAR ) ( as P - CAR ) which comprises in the engineered (CAR ) (as P -CAR ) which comprises in the engineered immune cell in combination with the N - CAR ; said CS1 immune cell in combination with the N -CAR ; said CS1 specific chimeric antigen receptor (CAR ) having one of the specific chimeric antigen receptor (CAR ) having one of the polypeptide structure selected from V1 to V6 , preferably V1, polypeptide structure selected from V1 to V6 , preferably V1 , V3 and V5 as illustrated in FIG . 1 , said structure comprising V3 and V5 as illustrated in FIG . 1 , said structure comprising an extra cellular ligand binding - domain comprising VH and an extra cellular ligand binding -domain comprising VH and VL from a monoclonal anti - CS1 antibody , a hinge , a trans US 2017 /0296623 A1 Oct . 19 , 2017 39 membrane domain and a cytoplasmic domain including a specifically the CS1 antigen , and an intracellular sig CD3 zeta signaling domain and a co - stimulatory domain naling domain , optionally with co - stimulatory domain . from 4 - 1BB . 10738 ) According to an embodiment, the isolated immune [ 0729 ] According to a preferred embodiment, the isolated cell includes at least a N -CAR which comprises at least : immune cell includes at least a N -CAR which comprises at 0739 ] an extracellular binding domain , which is able to least: bind to an “ off - target ” antigen on a healthy cell ; 10730 ] an extracellular binding domain , comprising a [0740 ] an inhibitory transmembrane polypeptide having polypeptide sequence sharing more than 80 % , prefer a sequence of SEQ ID NO : 1 (human KI2L2 ) , SEQ ID ably 90 % and more preferably 95 % and even more NO : 2 (human KI2L1) , SEQ ID NO : 3 (human preferably 100 % identity identity with SEQ ID NO : 92 FCG2B ) , SEQ ID NO : 4 (human KI2L3 ) , SEQ ID NO : (troponin C ) , SEQ ID NO : 93 (beta - 1 integrin ) , SEQ ID 5 (human KI3L2 ), SEQ ID NO : 6 (human KI2L4 ), SEQ NO : 94 (CCKBR antigen ), SEQ ID NO : 95 (GALR1 ID NO : 7 ( human KI3L1) , SEQ ID NO : 8 ( human antigen ) or SEQ ID NO : 96 (CUBN antigen ); and ; KIZLA ) , SEQ ID NO : 9 ( human MILR1) , SEQ ID [0731 ] an inhibitory transmembrane polypeptide having NO : 10 (human LIRB4 ) , SEQ ID NO : 11 (human a polypeptide sequence of SEQ ID NO : 33 (human LIRB3) , SEQ ID NO : 12 ( human KI3L3 ), SEQ ID TR10D ) , SEQ ID NO : 34 (human TR10A ) and SEQ ID NO : 15 (human LIRB5 ) , SEQ ID NO : 16 (human NO : 35 ( human TR10B ) ; LIRB2) , SEQ ID NO : 18 (human FCRL4) , SEQ ID [ 0732 ] and a CS1 specific chimeric antigen receptor NO : 23 (human FCRL5) , SEQ ID NO : 24 (human (CAR ) ( as P -CAR ) which comprises in the engineered FCRL2 ) , SEO ID NO : 25 (human FCRL1) , SEO ID immune cell in combination with the N -CAR ; said CS1 NO :27 (human FCRL3 ) , SEQ ID NO : 28 (human specific chimeric antigen receptor (CAR ) having one of the MPZL1) , SEQ ID NO : 29 ( human PILRA ) or SEQ ID polypeptide structure selected from V1 to V6 , preferably V1, NO : 30 (human PVR ); V3 and V5 as illustrated in FIG . 1 , said structure comprising [ 0741 ] and a ROR1 specific chimeric antigen receptor an extra cellular ligand binding - domain comprising VH and (CAR ) ( as P -CAR ) which comprises in the engineered immune cell in combination with the N - CAR ; said VL from a monoclonal anti- CS1 antibody , a hinge, a trans ROR1 specific chimeric antigen receptor (CAR ) con membrane domain and a cytoplasmic domain including a taining at least one transmembrane polypeptide which CD3 zeta signaling domain and a co - stimulatory domain includes at least one extra -binding domain recognizing from 4 - 1BB . specifically the CS1 antigen , and an intracellular sig [0733 ] Preferably , the above CS1 specific chimeric anti naling domain , optionally with co - stimulatory domain . gen receptor (CAR ) (as P -CAR ) comprises an extra cellular [ 0742 ] According to an embodiment, the isolated immune ligand binding -domain in which VH and VL chains derive cell includes at least a N - CAR which comprises at least : from a monoclonal anti- CS1 antibody , such as the murine [ 0743 ] an extracellular binding domain , comprising a scFv Luc63, Luc90 , Luc34 , LucX1 and LucX2 antibodies polypeptide sequence sharing more than 80 % , prefer ( such as described in WO2015121454A1 SEQ ID NO . 38 to ably 90 % and more preferably 95 % and even more 47 ) optionally humanized . preferably 100 % identity identity with SEQ ID NO : 92 [ 0734 ] According to an embodiment, the isolated immune ( troponin C ) , SEQ ID NO : 93 (beta - 1 integrin ) , SEQ ID cell includes at least a N -CAR which comprises at least: NO : 94 (CCKBR antigen ), SEQ ID NO : 95 (GALR1 [ 0735 ] an extracellular binding domain , which is able to antigen ) or SEQ ID NO : 96 (CUBN antigen ) ; bind to an “ off -target ” antigen on a healthy cell ; [0744 ] an inhibitory transmembrane polypeptide having [ 0736 ] an inhibitory transmembrane polypeptide having of more than 80 % , preferably 90 % and more preferably a sequence with more than 80 % , preferably 90 % and 95 % identity and even more preferably 100 % identity more preferably 95 % identity and even more preferably with SEQ ID NO : 1 (human KI2L2 ) , SEQ ID NO : 2 100 % identity with SEQ ID NO : 1 (human KI2L2) , (human KI2L1) , SEQ ID NO : 3 (human FCG2B ) , SEQ SEQ ID NO : 2 (human KI2L1 ) , SEQ ID NO :3 (human ID NO :4 (human KI2L3) , SEQ ID NO : 5 (human FCG2B ) , SEQ ID NO : 4 (human KI2L3) , SEQ ID NO : KI3L2 ) , SEQ ID NO : 6 ( human KI2L4 ) , SEQ ID NO : 7 5 ( human KI3L2 ) , SEQ ID NO : 6 ( human KI2L4 ) , SEO (human KI3L1 ) , SEQ ID NO : 8 (human KIZLA ) , SEQ ID NO : 7 (human KI3L1) , SEQ ID NO : 8 (human ID NO : 9 (human MILR1) , SEQ ID NO : 10 (human KIZLA ) , SEQ ID NO : 9 (human MILR1 ) , SEQ ID LIRB4 ) , SEQ ID NO : 11 (human LIRB3) , SEQ ID NO : 10 (human LIRB4 ) , SEQ ID NO : 11 (human NO : 12 ( human KI3L3 ), SEQ ID NO : 15 (human LIRB3 ) , SEQ ID NO : 12 (human KI3L3 ) , SEQ ID LIRB5 ) , SEQ ID NO : 16 (human LIRB2 ) , SEQ ID NO : 15 (human LIRB5 ) , SEQ ID NO : 16 (human NO : 18 ( human FCRL4 ) , SEQ ID NO : 23 (human LIRB2 ) , SEQ ID NO : 18 (human FCRL4 ) , SEQ ID FCRL5 ), SEQ ID NO :24 (human FCRL2) , SEQ ID NO : 23 (human FCRL5 ), SEQ ID NO : 24 (human NO : 25 (human FCRL1) , SEQ ID NO : 27 (human FCRL2 ), SEQ ID NO : 25 ( human FCRL1) , SEQ ID FCRL3 ) , SEQ ID NO : 28 (human MPZL1) , SEQ ID NO :27 (human FCRL3 ) , SEQ ID NO :28 (human NO : 29 (human PILRA ) or SEQ ID NO :30 (human MPZL1) , SEQ ID NO :29 (human PILRA ) or SEQ ID PVR ); NO : 30 ( human PVR ) ; [0745 ] and a ROR1 specific chimeric antigen receptor [ 0737 ] and a ROR1 specific chimeric antigen receptor (CAR ) ( as P -CAR ) which comprises in the engineered (CAR ) (as P -CAR ) which comprises in the engineered immune cell in combination with the N -CAR ; said ROR1 immune cell in combination with the N -CAR ; said specific chimeric antigen receptor (CAR ) having one of the ROR1 specific chimeric antigen receptor (CAR ) con polypeptide structure selected from V1 to V6 , preferably V1, taining at least one transmembrane polypeptide which V3 and V5 as illustrated in FIG . 1 , said structure comprising includes at least one extra -binding domain recognizing an extra cellular ligand binding -domain comprising VH and US 2017 /0296623 A1 Oct . 19, 2017 40 VL from a monoclonal anti -ROR1 antibody , a hinge, a [0757 ] and ; transmembrane domain and a cytoplasmic domain including [0758 ] an inhibitory transmembrane polypeptide having a CD3 zeta signaling domain and a co - stimulatory domain a polypeptide sequence of SEQ ID NO : 33 (human from 4 - 1BB . TR10D ) , SEQ ID NO : 34 (human TR10A ) and SEQ ID [ 0746 ] According to an embodiment, the isolated immune NO : 35 (human TR10B ) ; cell includes at least a N -CAR which comprises at least : [ 0759 ] and a ROR1 specific chimeric antigen receptor [0747 ] an extracellular binding domain , comprising a (CAR ) ( as P -CAR ) which comprises in the engineered polypeptide sequence sharing more than 80 % , prefer immune cell in combination with the N -CAR ; said ably 90 % and more preferably 95 % and even more ROR1 specific chimeric antigen receptor (CAR ) con preferably 100 % identity identity with SEQ ID NO : 92 taining at least one transmembrane polypeptide which (troponin C ) , SEQ ID NO : 93 (beta - 1 integrin ) , SEQ ID includes at least one extra -binding domain recognizing NO : 94 ( CCKBR antigen ), SEQ ID NO : 95 (GALR1 specifically the ROR1 antigen , and an intracellular antigen ) or SEQ ID NO : 96 (CUBN antigen ) ; signaling domain , optionally with co - stimulatory [0748 ] an inhibitory transmembrane polypeptide having domain . a of SEQ ID NO : 1 (human KI2L2) , SEQ ID NO : 2 [ 0760 ] According to a preferred embodiment, the isolated (human KI2L1 ), SEQ ID NO :3 (human FCG2B ), SEQ immune cell includes at least a N -CAR which comprises at ID NO : 4 (human KI2L3) , SEQ ID NO : 5 (human least : KI3L2 ) , SEQ ID NO : 6 ( human KI2L4 ) , SEQ ID NO : 7 [ 0761] an extracellular binding domain , comprising a ( human KI3L1) , SEQ ID NO : 8 (human KIZLA ) , SEQ polypeptide sequence sharing more than 80 % , prefer ID NO : 9 (human MILR1) , SEQ ID NO : 10 (human ably 90 % and more preferably 95 % and even more LIRB4 ) , SEO ID NO : 11 ( human LIRB3 ) , SEO ID preferably 100 % identity identity with SEQ ID NO : 92 NO : 12 ( human KI3L3) , SEQ ID NO : 15 human LIRB5 ), SEQ ID NO : 16 (human LIRB2 ), SEQ ID ( troponin C ), SEQ ID NO : 93 (beta - 1 integrin ), SEQ ID NO : 18 (human FCRL4 ), SEQ ID NO : 23 (human NO : 94 (CCKBR antigen ) , SEQ ID NO : 95 (GALR1 FCRL5 ) , SEQ ID NO : 24 (human FCRL2 ), SEQ ID antigen ) or SEQ ID NO : 96 (CUBN antigen ) ; and ; NO : 25 (human FCRL1) , SEQ ID NO : 27 (human [0762 ] an inhibitory transmembrane polypeptide having FCRL3 ) , SEQ ID NO : 28 (human MPZL1) , SEQ ID a polypeptide sequence of more than 80 % , preferably NO : 29 (human PILRA ) or SEO ID NO : 30 (human 90 % and more preferably 95 % identity and even more PVR ) ; preferably 100 % identity with SEQ ID NO : 33 (human [ 0749 ] and a ROR1 specific chimeric antigen receptor TR10D ), SEQ ID NO : 34 (human TR10A ) and SEQ ID (CAR ) ( as P -CAR ) which comprises in the engineered NO : 35 (human TR10B ) ; immune cell in combination with the N - CAR ; said ROR1 [0763 ] and a ROR1 specific chimeric antigen receptor specific chimeric antigen receptor (CAR ) having one of the (CAR ) (as P - CAR ) which comprises in the engineered polypeptide structure selected from V1 to V6 , preferably V1, immune cell in combination with the N -CAR ; said ROR1 V3 and V5 as illustrated in FIG . 1 , said structure comprising specific chimeric antigen receptor (CAR ) having one of the an extra cellular ligand binding - domain comprising VH and polypeptide structure selected from V1 to V6 , preferably V1, VL from a monoclonal anti -ROR1 antibody , a hinge , a V3 and V5 as illustrated in FIG . 1 , said structure comprising transmembrane domain and a cytoplasmic domain including an extra cellular ligand binding -domain comprising VH and a CD3 zeta signaling domain and a co -stimulatory domain VL from a monoclonal anti -ROR1 antibody , a hinge , a from 4 - 1BB . transmembrane domain and a cytoplasmic domain including [0750 ] According to an embodiment, the isolated immune a CD3 zeta signaling domain and a co - stimulatory domain cell includes at least a N -CAR which comprises at least : from 4 - 1BB . [ 0751 ] an extracellular binding domain , which is able to [0764 ] According to a preferred embodiment, the isolated bind to an “ off- target” antigen on a healthy cell ; immune cell includes at least a N -CAR which comprises at [ 0752 ] and ; least: [0753 ] an inhibitory transmembrane polypeptide having [ 0765 ] an extracellular binding domain , comprising a a polypeptide sequence of more than 80 % , preferably polypeptide sequence sharing more than 80 % , prefer 90 % and more preferably 95 % identity and even more ably 90 % and more preferably 95 % and even more preferably 100 % identity with SEQ ID NO : 33 (human preferably 100 % identity identity with SEQ ID NO : 92 TR10D ) , SEQ ID NO : 34 (human TR10A ) and SEQ ID ( troponin C ) , SEQ ID NO : 93 (beta - 1 integrin ) , SEO ID NO : 35 (human TR10B ) ; NO : 94 (CCKBR antigen ) , SEQ ID NO : 95 (GALR1 [ 0754 ] and a ROR1 specific chimeric antigen receptor antigen ) or SEQ ID NO : 96 (CUBN antigen ) ; and ; (CAR ) ( as P -CAR ) which comprises in the engineered [0766 ] an inhibitory transmembrane polypeptide having immune cell in combination with the N - CAR ; said a polypeptide sequence of SEQ ID NO : 33 (human ROR1 specific chimeric antigen receptor (CAR ) con TR10D ), SEQ ID NO : 34 ( human TR10A ) and SEQ ID taining at least one transmembrane polypeptide which NO : 35 ( human TR10B ) ; includes at least one extra - binding domain recognizing [0767 ] and a ROR1 specific chimeric antigen receptor specifically the ROR1 antigen , and an intracellular (CAR ) ( as P -CAR ) which comprises in the engineered signaling domain , optionally with co - stimulatory immune cell in combination with the N -CAR ; said ROR1 domain . specific chimeric antigen receptor (CAR ) having one of the [0755 ] According to an embodiment, the isolated immune polypeptide structure selected from V1 to V6 , preferably V1, cell includes at least a N -CAR which comprises at least : V3 and V5 as illustrated in FIG . 1 , said structure comprising [0756 ] an extracellular binding domain , which is able to an extra cellular ligand binding- domain comprising VH and bind to an “ off -target ” antigen on a healthy cell ; VL from a monoclonal anti -ROR1 antibody , a hinge , a US 2017 /0296623 A1 Oct . 19 , 2017 transmembrane domain and a cytoplasmic domain including an extra cellular ligand binding - domain comprising VH and a CD3 zeta signaling domain and a co - stimulatory domain VL from a monoclonal anti - CLL - 1 antibody , a hinge, a from 4 - 1BB . transmembrane domain and a cytoplasmic domain including [ 0768 ] According to an embodiment, the isolated immune a CD3 zeta signaling domain and a co - stimulatory domain cell includes at least a N -CAR which comprises at least: from 4 - 1BB . 10769 ] an extracellular binding domain , which binds to [0776 ] According to a preferred embodiment, the isolated an “ off - site ” cell surface antigen expressed in healthy immune cell includes at least a N -CAR which comprises at cells or in immune cells ; least : [0770 ] an inhibitory transmembrane polypeptide having [0777 ] an extracellular binding domain , which binds to a sequence with more than 80 % , preferably 90 % and an “ off -site ” cell surface antigen expressed in healthy more preferably 95 % identity and even more preferably cells or in immune cells ; 100 % identity with SEQ ID NO : 1 (human KI2L2) , [ 0778 ] an inhibitory transmembrane polypeptide having SEQ ID NO : 2 (human KI2L1) , SEQ ID NO : 3 ( human a polypeptide sequence ofmore than 80 % , preferably FCG2B ) , SEQ ID NO : 4 (human KI2L3 ), SEQ ID NO : 90 % and more preferably 95 % identity and even more 5 ( human KI3L2 ), SEQ ID NO :6 (human KI2L4 ) , SEQ preferably 100 % identity with SEQ ID NO : 33 (human ID NO : 7 (human KIZL1) , SEQ ID NO : 8 (human TR10D ), SEQ ID NO : 34 (human TR10A ) and SEQ ID KIZLA ), SEQ ID NO : 9 ( human MILR1) , SEQ ID NO : 35 (human TR10B ) ; NO : 10 (human LIRB4 ), SEQ ID NO : 11 (human [0779 ] and a CLL - 1 specific chimeric antigen receptor LIRB3 ) , SEQ ID NO : 12 (human KI3L3 ) , SEQ ID (CAR ) ( as P - CAR ) which comprises in the engineered NO :15 (human LIRB5) , SEQ ID NO : 16 (human immune cell in combination with the N - CAR ; said CLL - 1 LIRB2 ) , SEQ ID NO : 18 (human FCRL4 ) , SEQ ID specific chimeric antigen receptor (CAR ) having one of the NO :23 (human FCRL5 ) , SEQ ID NO : 24 (human polypeptide structure selected from V1 to V6 , preferably V1 , FCRL2) , SEQ ID NO : 25 (human FCRL1) , SEQ ID V3 and V5 as illustrated in FIG . 1 , said structure comprising NO : 27 (human FCRL3 ) , SEQ ID NO : 28 (human an extra cellular ligand binding -domain comprising VH and MPZL1) , SEQ ID NO : 29 (human PILRA ) or SEQ ID VL from a monoclonal anti- CLL - 1 antibody, a hinge , a NO : 30 ( human PVR ) ; transmembrane domain and a cytoplasmic domain including [0771 ] and a CLL - 1 specific chimeric antigen receptor a CD3 zeta signaling domain and a co - stimulatory domain (CAR ) ( as P - CAR ) which comprises in the engineered from 4 - 1BB . immune cell in combination with the N -CAR ; said CLL - 1 [0780 ] According to a preferred embodiment, the isolated specific chimeric antigen receptor (CAR ) having one of the immune cell includes at least a N -CAR which comprises at polypeptide structure selected from V1 to V6 , preferably V1, least : V3 and V5 as illustrated in FIG . 1 , said structure comprising [ 0781 ] an extracellular binding domain , which binds to an extra cellular ligand binding - domain comprising VH and an " off- site ” cell surface antigen expressed in healthy VL from a monoclonal anti -CLL - 1 antibody, a hinge , a cells or in immune cells ; transmembrane domain and a cytoplasmic domain including [0782 ] an inhibitory transmembrane polypeptide having a CD3 zeta signaling domain and a co -stimulatory domain a polypeptide sequence of SEQ ID NO : 33 (human from 4 - 1BB . TR10D ), SEQ ID NO : 34 ( human TR10A ) and SEQ ID [0772 ] According to an embodiment, the isolated immune NO : 35 (human TR10B ); cell includes at least a N -CAR which comprises at least : [0783 ] and a CLL - 1 specific chimeric antigen receptor [0773 ] an extracellular binding domain , which binds to (CAR ) ( as P - CAR ) which comprises in the engineered an " off - site ” cell surface antigen expressed in healthy immune cell in combination with the N -CAR ; said CLL - 1 cells or in immune cells; specific chimeric antigen receptor (CAR ) having one of the [0774 ] an inhibitory transmembrane polypeptide having polypeptide structure selected from V1 to V6 , preferably V1 , a sequence of SEQ ID NO : 1 (human KI2L2 ) , SEQ ID V3 and V5 as illustrated in FIG . 1 , said structure comprising NO : 2 (human KI2L1) , SEQ ID NO : 3 (human an extra cellular ligand binding - domain comprising VH and FCG2B ) , SEQ ID NO : 4 (human KI2L3) , SEQ ID NO : VL from a monoclonal anti -CLL - 1 antibody, a hinge , a 5 (human KI3L2) , SEQ ID NO : 6 (human KI2L4) , SEQ transmembrane domain and a cytoplasmic domain including ID NO : 7 (human KI3L1) , SEQ ID NO : 8 (human a CD3 zeta signaling domain and a co - stimulatory domain KI2LA ), SEQ ID NO : 9 ( human MILR1) , SEQ ID from 4 - 1BB . NO : 10 (human LIRB4 ) , SEQ ID NO : 11 (human [0784 ] As examples, said V? and Vh are preferably LIRB3 ) , SEQ ID NO : 12 ( human KI3L3 ) , SEQ ID selected from the antibodies referred to in the literature as NO : 15 (human LIRB5 ), SEQ ID NO : 16 (human SC002 -357 , SCO2 - 378 and SCO2- 161 in WO2005 /00894 LIRB2) , SEQ ID NO : 18 (human FCRL4 ) , SEQ ID ( Applicant: Crucell Holland BV ) ; M26 , M31 , G4, M22 , NO :23 (human FCRL5) , SEQ ID NO :24 (human M29, M2, M5, G12 in WO2013 /169625 ( Applicant: Celle FCRL2) , SEQ ID NO : 25 (human FCRL1) , SEQ ID rant Therapeutics ); and 21 .26 , 1075 . 7 in WO2009 /051974 NO :27 ( human FCRL3) , SEQ ID NO : 28 (human ( Applicant : Nuvelo Inc ) . MPZL1) , SEQ ID NO :29 (human PILRA ) or SEQ ID [0785 ] According to a preferred embodiment, the isolated NO :30 (human PVR ) ; immune cell includes at least a N -CAR which comprises at 10775 ] and a CLL - 1 specific chimeric antigen receptor least: (CAR ) ( as P -CAR ) which comprises in the engineered [0786 ] an extracellular binding domain , which is able to immune cell in combination with the N -CAR ; said CLL - 1 bind to an " off - target " antigen on a healthy or immune specific chimeric antigen receptor ( CAR ) having one of the cell and ; polypeptide structure selected from V1 to V6 , preferably V1 , [ 0787 ] an inhibitory transmembrane polypeptide having V3 and V5 as illustrated in FIG . 1 , said structure comprising a sequence consisting essentially of amino acids No US 2017 /0296623 A1 Oct . 19 , 2017 201 -375 from SEQ ID NO : 8 (human KIZLA ) , amino acids No 420 -598 from SEQ ID NO : 16 (human acids No 206 - 348 from SEQ ID NO : 2 (human LIRB2 ) , amino acids No 375 -515 from SEQ ID NO : 18 KIR2DL1) , amino acids No 206 - 348 from SEQ ID (human FCRL4 ) , amino acids No 753 - 977 from SEQ NO : 1 (human KIR2DL2 ) , amino acids No 206 -341 ID NO : 23 (human FCRL5) , amino acids No 388 -508 from SEO ID NO : 4 ( human KIR2DL3 ) , amino acids from SEQ ID NO : 24 (human FCRL2 ), amino acids No No 206 -348 from SEQ ID NO :2 (human KIR2DL1 ), 292 -429 from SEQ ID NO : 25 (human FCRL1) , amino amino acids No 203 - 377 from SEQ ID NO : 6 (human acids No 564 - 734 from SEQ ID NO :27 (human KIR2DL4 ) , amino acids No 301- 444 from SEQ ID FCRL3 ) , amino acids No 147 - 269 from SEQ ID NO : 28 NO : 7 (human KIR3DL1 ) , amino acids No 301- 455 (human MPZL1) , amino acids No 151 - 303 from SEQ from SEQ ID NO : 5 (human KIR3DL2 ) , amino acids ID NO : 29 (human PILRA ) , amino acids No 329 -417 No 204 - 310 from SEQ ID NO : 24 (human FRGR2B ) , from SEQ ID NO :30 (human PVR ), amino acids No amino acids No 214 - 343 from SEQ ID NO : 9 (human 229 - 325 from SEQ ID NO : 36 (human CD200 recep MILR1) , amino acids No 216 - 448 from SEQ ID NO : 10 torl ) , amino acids No 181 - 386 from SEO ID NO : 33 (human LIRB4) , amino acids No 420 -631 from SEQ (human TR10D ) , amino acids No 230 -468 from SEQ ID NO : 11 (human LIRB3 ), amino acids No 296 - 410 ID NO : 34 ( human TR10A ) or amino acids No 179 - 440 from SEQ ID NO : 12 (human KI3L3 ), amino acids No from SEQ ID NO : 35 ( human TR10B ) ; 419 - 590 from SEQ ID NO : 15 (human LIRB5 ) , amino [ 0792 ] and a P - CAR which comprises in the engineered acids No 420 - 598 from SEQ ID NO : 16 ( human immune cell in combination with the N -CAR ; said P -CAR LIRB2) , amino acids No 375 -515 from SEQ ID NO : 18 comprising one transmembrane polypeptide comprising at ( human FCRL4 ), amino acids No 753 - 977 from SEQ ID NO :23 (human FCRL5 ) , amino acids No 388 -508 least one extracellular ligand -binding domain able to bind to from SEO ID NO : 24 (human FCRL2 ) , amino acids No CD123 antigen , and one signal- transducing domain , option 292 - 429 from SEQ ID NO : 25 ( human FCRL1) , amino ally with a co - stimulatory domain . acids No 564 - 734 from SEQ ID NO :27 (human [0793 ] According to an embodiment, the isolated immune FCRL3 ) , amino acids No 147 - 269 from SEQ ID NO : 28 cell includes at least a N -CAR which comprises at least : (human MPZL1) , amino acids No 151- 303 from SEQ [ 0794 ] an extracellular binding domain , comprising a ID NO : 29 ( human PILRA ) , amino acids No 329 - 417 polypeptide sequence sharing more than 80 % , prefer from SEQ ID NO :30 ( human PVR ) , amino acids No ably 90 % and more preferably 95 % identity with SEQ 229 - 325 from SEQ ID NO : 36 (human CD200 recep ID NO : 81 (CD4 antigen ) , SEQ ID NO : 82 ( CD20 torl ) , amino acids No 181 - 386 from SEQ ID NO : 33 antigen ), SEQ ID NO : 83 ( CD22 antigen ) , SEQ ID NO : (human TR10D ) , amino acids No 230 - 468 from SEQ 84 (CD25 antigen ) or SEQ ID NO : 85 (MUC1 antigen ) ; ID NO :34 (human TR10A ) or amino acids No 179 - 440 [0795 ] an inhibitory transmembrane polypeptidehaving from SEQ ID NO : 35 (human TR10B ) ; a sequence consisting essentially of amino acids No [ 0788 ] and a P -CAR which comprises in the engineered 201 - 375 from SEQ ID NO : 8 ( human KIZLA ) , amino immune cell in combination with the N - CAR ; said P -CAR acids No 206 - 348 from SEQ ID NO : 2 (human comprising one transmembrane polypeptide comprising at KIR2DL1) , amino acids No 206 -348 from SEQ ID least one extracellular ligand - binding domain able to bind to NO : 1 ( human KIR2DL2 ) , amino acids No 206 - 341 CD123 antigen , and one signal- transducing domain , option from SEQ ID NO :4 (human KIR2DL3 ), amino acids ally with a co -stimulatory domain . No 206 - 348 from SEQ ID NO : 2 (human KIR2DL1) , [0789 ] According to another preferred embodiment, the amino acids No 203 -377 from SEQ ID NO :6 (human isolated immune cell includes at least a N -CAR which KIR2DL4 ), amino acids No 301 -444 from SEQ ID comprises at least : NO : 7 ( human KIR3DL1) , amino acids No 301- 455 [0790 ] an extracellular binding domain , which is able to from SEQ ID NO :5 (human KIR3DL2 ), amino acids bind to an “ off - target ” antigen on a healthy or immune No 204 - 310 from SEQ ID NO : 24 (human FRGR2B ) , cell and ; amino acids No 214 - 343 from SEQ ID NO : 9 (human [0791 ] an inhibitory transmembrane polypeptide having MILR1 ) , amino acids No 216 -448 from SEO ID NO : 10 a sequence consisting essentially of amino acids No (human LIRB4 ) , amino acids No 420 -631 from SEQ 201- 375 from SEQ ID NO : 8 (human KIZLA ), amino ID NO : 11 (human LIRB3) , amino acids No 296 -410 acids No 206 - 348 from SEO ID NO : 2 (human from SEO ID NO : 12 (human KI3L3 ) , amino acids No KIR2DL1) , amino acids No 206 -348 from SEQ ID 419 - 590 from SEQ ID NO : 15 (human LIRB5 ) , amino NO : 1 (human KIR2DL2 ), amino acids No 206 - 341 acids No 420 - 598 from SEQ ID NO : 16 (human from SEQ ID NO : 4 (human KIR2DL3 ) , amino acids LIRB2 ) , amino acids No 375 -515 from SEO ID NO : 18 No 206 - 348 from SEQ ID NO : 2 (human KIR2DL1 ) , (human FCRL4) , amino acids No 753 - 977 from SEQ amino acids No 203 - 377 from SEQ ID NO :6 (human ID NO :23 (human FCRL5) , amino acids No 388 -508 KIR2DL4 ), amino acids No 301 -444 from SEQ ID from SEQ ID NO : 24 (human FCRL2) , amino acids No NO : 7 (human KIR3DL1) , amino acids No 301 -455 292 - 429 from SEQ ID NO : 25 (human FCRL1) , amino from SEQ ID NO : 5 (human KIR3DL2 ) , amino acids acids No 564 - 734 from SEO ID NO : 27 ( human No 204 - 310 from SEQ ID NO : 24 (human FRGR2B ) , FCRL3) , amino acids No 147 - 269 from SEQ ID NO : 28 amino acids No 214 - 343 from SEQ ID NO : 9 (human ( human MPZL1 ) , amino acids No 151 -303 from SEO MILR1 ) , amino acids No 216 - 448 from SEO ID NO : 10 ID NO : 29 (human PILRA ) , amino acids No 329 -417 (human LIRB4) , amino acids No 420 -631 from SEQ from SEQ ID NO : 30 (human PVR ), amino acids No ID NO : 11 (human LIRB3 ) , amino acids No 296 - 410 229 - 325 from SEQ ID NO :36 ( human CD200 recep from SEQ ID NO : 12 (human KI3L3 ), amino acids No torl ) , amino acids No 181 - 386 from SEQ ID NO :33 419 - 590 from SEQ ID NO : 15 (human LIRB5 ) , amino (human TR10D ) , amino acids No 230 -468 from SEQ US 2017 /0296623 A1 Oct . 19 , 2017 43 ID NO : 34 ( human TR10A ) or amino acids No 179 - 440 [0802 ] an extracellular binding domain ,which is able to from SEQ ID NO : 35 (human TR10B ) ; bind to an “ off - target” antigen on a healthy or immune [0796 ] and a P -CAR which comprises in the engineered cell and ; immune cell in combination with the N -CAR ; said P -CAR [0803 ] an inhibitory transmembrane polypeptide having comprising one transmembrane polypeptide comprising at a polypeptide sequence of SEQ ID NO : 33 (human least one extracellular ligand -binding domain able to bind to TR10D ), SEQ ID NO : 34 (human TR10A ) and SEQ ID CD123 antigen , and one signal- transducing domain , option NO : 35 (human TR10B ) . ally with a co - stimulatory domain . [0804 ] and a P -CAR which comprises in the engineered immune cell in combination with the N -CAR ; said P -CAR [ 0797 ] According to another preferred embodiment, the comprising one transmembrane polypeptide comprising at isolated immune cell includes at least a N -CAR which least one extracellular ligand - binding domain able to bind to comprises at least : CD123 antigen , and one signal- transducing domain , option [0798 ] an extracellular binding domain , comprising a ally with a co - stimulatory domain . polypeptide sequence sharing more than 80 % , prefer ably 90 % and more preferably 95 % identity with SEQ [0805 ] According to another preferred embodiment, the ID NO : XX (CD4 antigen ) , SEQ ID NO : XX (CD20 isolated immune cell includes at least a N -CAR which antigen ), SEQ ID NO : XX (CD22 antigen ), SEQ ID comprises at least: NO : XX (CD25 antigen ) or SEQ ID NO : XX (MUC1 [ 0806 ] an extracellular binding domain , which is able to antigen ); bind to an “ off - target” antigen on a healthy or immune cell and ; [0799 ] an inhibitory transmembrane polypeptide having [0807 ] an inhibitory transmembrane polypeptide having a sequence consisting essentially of amino acids No a sequence consisting essentially of amino acids No 201- 375 from SEQ ID NO : 8 (human KIZLA ), amino 201 -375 from SEQ ID NO : 8 ( human KIZLA ) , amino acids No 206 - 348 from SEQ ID NO : 2 (human acids No 206 -348 from SEQ ID NO : 2 (human KIR2DL1) , amino acids No 206 -348 from SEQ ID KIR2DL1 ) , amino acids No 206 -348 from SEQ ID NO : 1 (human KIR2DL2 ) , amino acids No 206 - 341 NO : 1 ( human KIR2DL2) , amino acids No 206 - 341 from SEQ ID NO : 4 (human KIR2DL3) , amino acids from SEQ ID NO : 4 (human KIR2DL3 ) , amino acids No 206 - 348 from SEQ ID NO : 2 ( human KIR2DL1) , No 206 - 348 from SEQ ID NO : 2 (human KIR2DL1) , amino acids No 203 - 377 from SEQ ID NO : 6 (human amino acids No 203 -377 from SEQ ID NO :6 (human KIR2DL4 ), amino acids No 301 - 444 from SEQ ID KIR2DL4 ) , amino acids No 301- 444 from SEQ ID NO : 7 (human KIR3DL1) , amino acids No 301- 455 NO : 7 ( human KIR3DL1) , amino acids No 301 -455 from SEQ ID NO : 5 (human KIR3DL2 ) , amino acids from SEQ ID NO : 5 (human KIR3DL2 ), amino acids No 204 - 310 from SEQ ID NO : 24 (human FRGR2B ) , No 204 - 310 from SEQ ID NO :24 (human FRGR2B ), amino acids No 214 - 343 from SEQ ID NO : 9 (human amino acids No 214 -343 from SEQ ID NO : 9 (human MILR1) , amino acids No 216 - 448 from SEQ ID NO : 10 MILR1) , amino acids No 216 -448 from SEQ ID NO : 10 (human LIRB4 ), amino acids No 420 -631 from SEQ (human LIRB4 ), amino acids No 420 -631 from SEQ ID NO : 11 ( human LIRB3) , amino acids No 296 -410 ID NO : 11 (human LIRB3 ) , amino acids No 296 -410 from SEO ID NO : 12 ( human KI3L3 ) , amino acids No from SEQ ID NO : 12 (human KI3L3 ), amino acids No 419 - 590 from SEQ ID NO : 15 (human LIRB5 ), amino 419 -590 from SEQ ID NO : 15 (human LIRB5) , amino acids No 420 - 598 from SEQ ID NO : 16 (human acids No 420 - 598 from SEQ ID NO : 16 (human LIRB2) , amino acids No 375 - 515 from SEQ ID NO : 18 LIRB2) , amino acids No 375 -515 from SEQ ID NO : 18 ( human FCRL4 ) , amino acids No 753 - 977 from SEQ (human FCRL4 ) , amino acids No 753 - 977 from SEO ID NO : 23 (human FCRL5 ) , amino acids No 388 - 508 ID NO : 23 (human FCRL5 ) , amino acids No 388 - 508 from SEQ ID NO :24 (human FCRL2) , amino acids No from SEQ ID NO :24 (human FCRL2 ) , amino acids No 292 - 429 from SEQ ID NO : 25 (human FCRL1 ), amino 292 -429 from SEQ ID NO : 25 (human FCRL1) , amino acids No 564 - 734 from SEQ ID NO : 27 (human acids No 564 - 734 from SEQ ID NO : 27 (human FCRL3) , amino acids No 147 - 269 from SEQ ID NO : 28 FCRL3 ) , amino acids No 147 - 269 from SEQ ID NO : 28 (human MPZL1) , amino acids No 151 - 303 from SEO (human MPZL1) , amino acids No 151- 303 from SEQ ID NO : 29 (human PILRA ) , amino acids No 329 -417 ID NO : 29 (human PILRA ) , amino acids No 329 - 417 from SEQ ID NO : 30 (human PVR ), amino acids No from SEQ ID NO : 30 ( human PVR ) , amino acids No 229 - 325 from SEQ ID NO : 36 (human CD200 recep 229 - 325 from SEQ ID NO : 36 (human CD200 recep tori) , amino acids No 181- 386 from SEQ ID NO : 33 torl) , amino acids No 181 -386 from SEQ ID NO :33 ( human TR10D ) , amino acids No 230 - 468 from SEQ (human TR10D ) , amino acids No 230 - 468 from SEQ ID NO : 34 (human TR10A ) or amino acids No 179 -440 ID NO :34 (human TR10A ) or amino acids No 179 -440 from SEQ ID NO :35 (human TR10B ) ; from SEQ ID NO :35 (human TR10B ) ; [ 0800 ] and a P -CAR which comprises in the engineered [0808 ] and a P -CAR which comprises in the engineered immune cell in combination with the N -CAR ; said P -CAR immune cell in combination with the N -CAR ; said P -CAR comprising one transmembrane polypeptide comprising at comprising one transmembrane polypeptide comprising at least one extracellular ligand -binding domain able to bind to least one extracellular ligand -binding domain able to bind to CD123 antigen , and one signal -transducing domain , option CD123 antigen , and one signal- transducing domain , option ally with a co - stimulatory domain . ally with a co - stimulatory domain . [ 0801 ] According to a preferred embodiment, the isolated [0809 ] According to a preferred embodiment, the isolated immune cell includes at least a N -CAR which comprises at immune cell includes at least a N -CAR which comprises at least: least: US 2017 /0296623 A1 Oct . 19 , 2017 44 [ 0810 ] an extracellular binding domain , comprising a antigen ), SEQ ID NO : XX (CD22 antigen ), SEQ ID polypeptide sequence sharing more than 80 % , prefer NO : XX (CD25 antigen ) or SEQ ID NO : XX (MUC1 ably 90 % and more preferably 95 % identity with SEQ antigen ); ID NO : XX ( CD4 antigen ) , SEQ ID NO : XX (CD20 [ 0816 ] an inhibitory transmembrane polypeptide having antigen ) , SEQ ID NO : XX (CD22 antigen ) , SEQ ID a polypeptide sequence of more than 80 % , preferably NO : XX ( CD25 antigen ) or SEQ ID NO : XX (MUC1 90 % and more preferably 95 % identity with SEQ ID antigen ) NO : 33 (human TR10D ), SEQ ID NO : 34 (human TR10A ) and SEQ ID NO : 35 ( human TR10B ) ; [ 0811 ] and ; 10817 ] and a P -CAR which comprises in the engineered [0812 ] an inhibitory transmembrane polypeptide having immune cell in combination with the N - CAR ; said P -CAR a sequence consisting essentially of amino acids No comprising one transmembrane polypeptide comprising at 201 - 375 from SEQ ID NO :8 (human KIZLA ) , amino least one extracellular ligand -binding domain able to bind to acids No 206 - 348 from SEQ ID NO : 2 (human CD123 antigen , and one signal- transducing domain , option KIR2DL1) , amino acids No 206 - 348 from SEQ ID ally with a co - stimulatory domain . NO : 1 (human KIR2DL2 ), amino acids No 206 - 341 (0818 ] According to a preferred embodiment, the isolated from SEQ ID NO : 4 (human KIR2DL3 ) , amino acids immune cell includes at least a N -CAR which comprises at No 206 -348 from SEQ ID NO : 2 (human KIR2DL1) , least : amino acids No 203 - 377 from SEQ ID NO :6 (human [0819 ] an extracellular binding domain , comprising a KIR2DL4 ) , amino acids No 301- 444 from SEQ ID polypeptide sequence sharing more than 80 % , prefer NO : 7 (human KIR3DL1) , amino acids No 301- 455 ably 90 % and more preferably 95 % identity with SEQ from SEQ ID NO : 5 (human KIR3DL2 ) , amino acids ID NO : XX (CD4 antigen ) , SEQ ID NO : XX (CD20 No 204 -310 from SEQ ID NO : 24 (human FRGR2B ) , antigen ) , SEQ ID NO : XX (CD22 antigen ) , SEQ ID amino acids No 214 - 343 from SEQ ID NO : 9 ( human NO : XX (CD25 antigen ) or SEQ ID NO : XX (MUC1 MILR1 ) , amino acids No 216 -448 from SEQ ID NO : 10 antigen ) ; (human LIRB4 ) , amino acids No 420 -631 from SEQ 10820 ] an inhibitory transmembrane polypeptide having ID NO : 11 (human LIRB3 ) , amino acids No 296 - 410 a polypeptide sequence of SEQ ID NO : 33 (human from SEO ID NO : 12 (human KI3L3 ) , amino acids No TR10D ) , SEQ ID NO : 34 (human TR10A ) and SEQ ID 419 -590 from SEQ ID NO : 15 ( human LIRB5 ) , amino NO : 35 (human TR10B ) . acids No 420 - 598 from SEQ ID NO : 16 (human [ 0821 ] and a P -CAR which comprises in the engineered LIRB2) , amino acids No 375 - 515 from SEQ ID NO : 18 immune cell in combination with the N - CAR ; said P -CAR (human FCRL4 ) , amino acids No 753 - 977 from SEQ comprising one transmembrane polypeptide comprising at ID NO : 23 (human FCRL5 ) , amino acids No 388 - 508 least one extracellular ligand -binding domain able to bind to from SEQ ID NO : 24 (human FCRL2 ) , amino acids No CD123 antigen , and one signal- transducing domain , option 292 - 429 from SEQ ID NO : 25 (human FCRL1) , amino ally with a co - stimulatory domain . acids No 564 - 734 from SEQ ID NO : 27 (human [0822 ] According to a more preferred embodiment, the FCRL3 ) , amino acids No 147 - 269 from SEQ ID NO : 28 isolated immune cell includes at least a N - CAR which ( human MPZL1) , amino acids No 151 - 303 from SEQ comprises at least : ID NO :29 (human PILRA ) , amino acids No 329 - 417 [0823 ] an extracellular binding domain , comprising a from SEQ ID NO : 30 ( human PVR ), amino acids No polypeptide sequence sharing more than 80 % , prefer 229 - 325 from SEQ ID NO : 36 (human CD200 recep ably 90 % and more preferably 95 % identity with SEQ torl ), amino acids No 181 - 386 from SEQ ID NO : 33 ID NO : XX (CD4 antigen ) , SEQ ID NO : XX (CD20 ( human TR10D ), amino acids No 230 - 468 from SEQ antigen ), SEQ ID NO : XX (CD22 antigen ) , SEQ ID ID NO : 34 ( human TR10A ) or amino acids No 179 -440 NO : XX (CD25 antigen ) or SEQ ID NO : XX (MUC1 from SEQ ID NO : 35 (human TR10B ) ; antigen ) [ 0813 ] and a CD123 specific chimeric antigen receptor (0824 ] and ; (CAR ) (as P -CAR ) which comprises in the engineered [0825 ] an inhibitory transmembrane polypeptide having immune cell in combination with the N -CAR ; said a polypeptide sequence of more than 80 % , preferably CD123 specific chimeric antigen receptor (CAR ) hav 90 % and more preferably 95 % identity with SEQ ID ing one of the polypeptide structure selected from V1 NO : 33 (human TR10D ), SEQ ID NO : 34 (human to V6 , preferably V1 , V3 and V5 as illustrated in FIG . TR10A ) and SEQ ID NO : 35 (human TR10B ) ; 1 , said structure comprising an extra cellular ligand [0826 ] and a CD123 specific chimeric antigen receptor binding- domain comprising VH and VL from a mono (CAR ) ( as P - CAR ) which comprises in the engineered clonal anti- CD123 antibody, a hinge, a transmembrane immune cell in combination with the N - CAR ; said CD123 domain and a cytoplasmic domain including a CD3 specific chimeric antigen receptor (CAR ) having one of the zeta signaling domain and a co - stimulatory domain polypeptide structure selected from V1 to V6 , preferably V1, from 4 - 1BB . V3 and V5 as illustrated in FIG . 1 , said structure comprising an extra cellular ligand binding -domain comprising VH and [0814 ] According to a preferred embodiment, the isolated VL from a monoclonal anti - CD123 antibody , a hinge, a immune cell includes at least a N -CAR which comprises at transmembrane domain and a cytoplasmic domain including least : a CD3 zeta signaling domain and a co - stimulatory domain [ 0815 ] an extracellular binding domain , comprising a from 4 - 1BB polypeptide sequence sharing more than 80 % , prefer [0827 ] Preferably , the above anti - CD123 CARs ( P -CARs ) ably 90 % and more preferably 95 % identity with SEQ having one of the polypeptide structure selected from V1, ID NO : XX ( CD4 antigen ) , SEQ ID NO : XX (CD20 V3 and V5 , as illustrated in FIG . 1 , said structure comprising US 2017 /0296623 A1 Oct . 19 , 2017 45 an extra cellular ligand binding -domain comprising VH and [0833 ] According to an embodiment, the isolated immune VL from a monoclonal anti - CD123 antibody, a hingea, cell includes at least a N -CAR which comprises at least: transmembrane domain , a cytoplasmic domain including a [ 0834 ] an extracellular binding domain , which is able to CD3 zeta signaling domain and a co - stimulatory domain bind to an “ off -target ” antigen on a healthy cell ; from 4 - 1BB , said 123 CAR having at least 80 % sequence [0835 ] and ; identity with either SEQ ID NO . 42 , SEQ ID NO . 44 or SEQ [0836 ] an inhibitory transmembrane polypeptide having ID NO . 46 . a sequence consisting essentially of amino acids No [0828 ] According to an embodiment, the isolated immune 201 - 375 from SEQ ID NO : 8 (human KIZLA ) , amino cell includes at least a N -CAR which comprises at least: acids No 206 - 348 from SEQ ID NO : 2 (human [0829 ] an extracellular binding domain , comprising a KIR2DL1 ), amino acids No 206 - 348 from SEQ ID polypeptide sequence sharing more than 80 % , prefer NO : 1 (human KIR2DL2 ) , amino acids No 206 - 341 ably 90 % and more preferably 95 % identity with SEQ from SEQ ID NO :4 (human KIR2DL3 ), amino acids ID NO : XX ( CD56 antigen ) , SEQ ID NO : XX ( CD205 No 206 - 348 from SEQ ID NO : 2 (human KIR2DL1) , antigen ) , SEQ ID NO : XX (CD83 antigen ), SEQ ID amino acids No 203 - 377 from SEQ ID NO : 6 (human NO : XX (CD206 antigen ), SEQ ID NO : XX (CD200 KIR2DL4 ), amino acids No 301 - 444 from SEQ ID antigen ), or SEQ ID NO : 8 (CD36 antigen ); NO : 7 ( human KIRGDL1 ) , amino acids No 301 - 455 10830 ] and ; from SEQ ID NO :5 (human KIR3DL2 ), amino acids [0831 ] an inhibitory transmembrane polypeptide having No 204 - 310 from SEQ ID NO : 24 (human FRGR2B ) , a sequence consisting essentially of amino acids No amino acids No 214 - 343 from SEQ ID NO : 9 (human 201- 375 from SEQ ID NO : 8 ( human KI2LA ) , amino MILR1) , amino acids No 216 -448 from SEQ ID NO : 10 acids No 206 - 348 from SEQ ID NO : 2 (human (human LIRB4 ) , amino acids No 420 -631 from SEQ KIR2DL1 ), amino acids No 206 - 348 from SEQ ID ID NO : 11 ( human LIRB3 ) , amino acids No 296 -410 NO : 1 (human KIR2DL2 ) , amino acids No 206 - 341 from SEQ ID NO : 12 (human KI3L3 ) , amino acids No from SEQ ID NO :4 (human KIR2DL3 ), amino acids 419 -590 from SEQ ID NO : 15 (human LIRB5 ) , amino No 206 -348 from SEQ ID NO : 2 ( human KIR2DL1) , acids No 420 -598 from SEQ ID NO : 16 (human amino acids No 203 - 377 from SEQ ID NO : 6 ( human LIRB2 ) , amino acids No 375 -515 from SEQ ID NO : 18 KIR2DL4 ) , amino acids No 301- 444 from SEQ ID (human FCRL4 ), amino acids No 753 - 977 from SEQ NO : 7 (human KIR3DL1) , amino acids No 301 -455 ID NO :23 (human FCRL5) , amino acids No 388 - 508 from SEQ ID NO :5 (human KIR3DL2 ), amino acids from SEO ID NO : 24 ( human FCRL2 ) , amino acids No No 204 -310 from SEQ ID NO : 24 (human FRGR2B ) , 292 - 429 from SEQ ID NO : 25 (human FCRL1 ) , amino amino acids No 214 - 343 from SEQ ID NO : 9 (human acids No 564 - 734 from SEQ ID NO : 27 (human MILR1) , amino acids No 216 - 448 from SEQ ID NO : 10 FCRL3 ), amino acids No 147 -269 from SEQ ID NO :28 ( human LIRB4 ) , amino acids No 420 -631 from SEQ (human MPZL1 ) , amino acids No 151 -303 from SEQ ID NO : 11 ( human LIRB3 ) , amino acids No 296 -410 ID NO : 29 (human PILRA ) , amino acids No 329 - 417 from SEQ ID NO : 12 (human KI3L3 ) , amino acids No from SEQ ID NO :30 (human PVR ), amino acids No 419 -590 from SEO ID NO : 15 ( human LIRB5 ) , amino 229 - 325 from SEQ ID NO : 36 ( human CD200 recep acids No 420 -598 from SEQ ID NO :16 (human tori ), amino acids No 181 -386 from SEQ ID NO :33 LIRB2 ) , amino acids No 375 -515 from SEO ID NO : 18 (human TR10D ) , amino acids No 230 - 468 from SEQ (human FCRL4) , amino acids No 753 - 977 from SEQ ID NO : 34 (human TR10A ) or amino acids No 179 - 440 ID NO : 23 (human FCRL5 ), amino acids No 388 -508 from SEQ ID NO :35 (human TR10B ) ; from SEQ ID NO :24 (human FCRL2) , amino acids No [0837 ] and a CD38 specific chimeric antigen receptor 292 - 429 from SEQ ID NO : 25 (human FCRL1 ) , amino (CAR ) (as P - CAR ) which comprises in the engineered acids No 564 - 734 from SEO ID NO : 27 ( human immune cell in combination with the N - CAR ; said CD38 FCRL3) , amino acids No 147 - 269 from SEQ ID NO : 28 specific chimeric antigen receptor (CAR ) containing at least (human MPZL1) , amino acids No 151 - 303 from SEQ one transmembrane polypeptide which includes at least one ID NO : 29 ( human PILRA ) , amino acids No 329 - 417 extra -binding domain recognizing specifically the CD38 from SEQ ID NO : 30 (human PVR ), amino acids No antigen , and an intracellular signaling domain , optionally 229 - 325 from SEO ID NO : 36 (human CD200 recep with co -stimulatory domain . torl ), amino acids No 181 - 386 from SEQ ID NO : 33 [0838 ) According to an embodiment, the isolated immune (human TR10D ) , amino acids No 230 -468 from SEQ cell includes at least a N - CAR which comprises at least: ID NO :34 (human TR10A ) or amino acids No 179 - 440 [0839 ] an extracellular binding domain , which is able to from SEQ ID NO : 35 ( human TR10B ) ; bind to an “ off -target ” antigen on a healthy cell ; [ 0832 ] and a CD123 specific chimeric antigen receptor [ 0840 ] and ; (CAR ) ( as P -CAR ) which comprises in the engineered [0841 ] an inhibitory transmembrane polypeptidehaving immune cell in combination with the N - CAR ; said a sequence consisting essentially of amino acids No CD123 specific chimeric antigen receptor (CAR ) hav 201 - 375 from SEQ ID NO : 8 ( human KIZLA ) , amino ing one of the polypeptide structure selected from V1 acids No 206 - 348 from SEQ ID NO : 2 (human to V6 , preferably V1, V3 and V5 as illustrated in FIG . KIR2DL1) , amino acids No 206 -348 from SEQ ID 1 , said structure comprising an extra cellular ligand NO : 1 (human KIR2DL2) , amino acids No 206 - 341 binding - domain comprising VH and VL from a mono from SEQ ID NO : 4 (human KIR2DL3 ) , amino acids clonal anti -CD123 antibody , a hinge , a transmembrane No 206 -348 from SEQ ID NO :2 (human KIR2DL1) , domain and a cytoplasmic domain including a CD3 amino acids No 203 - 377 from SEQ ID NO : 6 (human zeta signaling domain and a co - stimulatory domain KIR2DL4 ) , amino acids No 301- 444 from SEQ ID from 4 - 1BB . NO : 7 (human KIR3DL1) , amino acids No 301 -455 US 2017 /0296623 A1 Oct. 19 , 2017 46 from SEQ ID NO : 5 (human KIR3DL2) , amino acids FCRL3) , amino acids No 147 - 269 from SEQ ID NO : 28 No 204 - 310 from SEQ ID NO : 24 (human FRGR2B ) , (human MPZL1) , amino acids No 151 - 303 from SEQ amino acids No 214 - 343 from SEQ ID NO : 9 (human ID NO : 29 (human PILRA ) , amino acids No 329 -417 MILR1) , amino acids No 216 - 448 from SEQ ID NO : 10 from SEQ ID NO :30 (human PVR ), amino acids No ( human LIRB4 ) , amino acids No 420 -631 from SEQ 229 - 325 from SEQ ID NO : 36 (human CD200 recep ID NO : 11 ( human LIRB3 ) , amino acids No 296 -410 tori ), amino acids No 181 -386 from SEQ ID NO :33 from SEQ ID NO : 12 (human KI3L3 ) , amino acids No ( human TR10D ) , amino acids No 230 -468 from SEQ 419 -590 from SEQ ID NO : 15 (human LIRB5 ) , amino ID NO :34 (human TR10A ) or amino acids No 179 -440 acids No 420 - 598 from SEQ ID NO : 16 (human from SEQ ID NO : 35 (human TR10B ); LIRB2 ), amino acids No 375 -515 from SEQ ID NO : 18 [ 0846 ] and a CD19 specific chimeric antigen receptor (human FCRL4) , amino acids No 753 - 977 from SEQ (CAR ) (as P -CAR ) which comprises in the engineered ID NO : 23 (human FCRL5 ) , amino acids No 388 -508 immune cell in combination with the N -CAR ; said CD19 from SEQ ID NO :24 (human FCRL2) , amino acids No specific chimeric antigen receptor ( CAR ) having one of the 292 -429 from SEQ ID NO : 25 (human FCRL1) , amino polypeptide structure selected from V1 to V6 , preferably V1, acids No 564 - 734 from SEQ ID NO : 27 (human V3 and V5 as illustrated in FIG . 1 , said structure comprising FCRL3) , amino acids No 147 - 269 from SEQ ID NO : 28 an extra cellular ligand binding -domain comprising VH and ( human MPZL1 ) , amino acids No 151 - 303 from SEQ VL from a monoclonal anti - CD19 antibody, a hinge , a ID NO : 29 (human PILRA ) , amino acids No 329 -417 transmembrane domain and a cytoplasmic domain including from SEQ ID NO : 30 (human PVR ) , amino acids No a CD3 zeta signaling domain and a co - stimulatory domain 229 - 325 from SEQ ID NO : 36 (human CD200 recep from 4 - 1BB . torl) , amino acids No 181 - 386 from SEQ ID NO : 33 [0847 ] According to a preferred embodiment, the isolated (human TR10D ) , amino acids No 230 -468 from SEQ immune cell includes at least a N -CAR which comprises at ID NO :34 (human TR10A ) or amino acids No 179 -440 least: from SEQ ID NO : 35 (human TR10B ) ; [0848 ] an extracellular binding domain , which is able to [ 0842] and a CD38 specific chimeric antigen receptor bind to an “ off -target ” antigen on a healthy cell; (CAR ) (as P -CAR ) which comprises in the engineered [0849 ] and ; immune cell in combination with the N -CAR ; said CD38 [0850 ] an inhibitory transmembrane polypeptide having specific chimeric antigen receptor (CAR ) containing at least a sequence consisting essentially of amino acids No one transmembrane polypeptide which includes at least one 201 -375 from SEQ ID NO : 8 (human KIZLA ) , amino extra -binding domain recognizing specifically the CD38 acids No 206 - 348 from SEQ ID NO : 2 (human antigen , and an intracellular signaling domain , optionally KIR2DL1) , amino acids No 206 -348 from SEQ ID with co - stimulatory domain . NO : 1 (human KIR2DL2 ), amino acids No 206 - 341 [ 0843] According to a preferred embodiment, the isolated from SEQ ID NO : 4 (human KIR2DL3 ) , amino acids immune cell includes at least a N - CAR which comprises at No 206 -348 from SEQ ID NO : 2 (human KIR2DL1) , least : amino acids No 203 -377 from SEQ ID NO :6 (human 0844 ] an extracellular binding domain , which is able to KIR2DL4 ) , amino acids No 301 - 444 from SEO ID bind to an " off - target” antigen on a healthy or immune NO : 7 ( human KIR3DL1) , amino acids No 301- 455 cell ; from SEO ID NO : 5 (human KIR3DL2 ) , amino acids [ 0845 ] an inhibitory transmembrane polypeptide having No 204 - 310 from SEQ ID NO : 24 ( human FRGR2B ) , a sequence consisting essentially of amino acids No amino acids No 214 - 343 from SEQ ID NO : 9 (human 201- 375 from SEQ ID NO :8 (human KIZLA ), amino MILR1) , amino acids No 216 - 448 from SEO ID NO : 10 acids No 206 - 348 from SEQ ID NO : 2 (human (human LIRB4 ) , amino acids No 420 -631 from SEQ KIR2DL1) , amino acids No 206 - 348 from SEQ ID ID NO : 11 (human LIRB3 ) , amino acids No 296 - 410 NO : 1 (human KIR2DL2 ) , amino acids No 206 - 341 from SEQ ID NO : 12 (human KI3L3 ), amino acids No from SEQ ID NO :4 (human KIR2DL3 ), amino acids 419 -590 from SEQ ID NO : 15 (human LIRB5) , amino No 206 - 348 from SEQ ID NO : 2 (human KIR2DL1 ) , acids No 420 - 598 from SEQ ID NO : 16 ( human amino acids No 203 - 377 from SEQ ID NO : 6 ( human LIRB2 ) , amino acids No 375 -515 from SEO ID NO : 18 KIR2DL4 ) , amino acids No 301- 444 from SEQ ID (human FCRL4 ), amino acids No 753 - 977 from SEQ NO : 7 (human KIR3DL1) , amino acids No 301 - 455 ID NO : 23 (human FCRL5 ) , amino acids No 388 - 508 from SEQ ID NO :5 (human KIR3DL2 ), amino acids from SEQ ID NO :24 (human FCRL2 ) , amino acids No No 204 -310 from SEQ ID NO :24 (human FRGR2B ) , 292 - 429 from SEQ ID NO : 25 (human FCRL1) , amino amino acids No 214 - 343 from SEQ ID NO : 9 ( human acids No 564 - 734 from SEQ ID NO :27 (human MILR1) , amino acids No 216 - 448 from SEQ ID NO : 10 FCRL3 ) , amino acids No 147 -269 from SEQ ID NO :28 ( human LIRB4 ) , amino acids No 420 -631 from SEQ (human MPZL1) , amino acids No 151- 303 from SEQ ID NO : 11 (human LIRB3 ), amino acids No 296 -410 ID NO : 29 (human PILRA ) , amino acids No 329 -417 from SEO ID NO : 12 ( human KI3L3 ) , amino acids No from SEQ ID NO :30 (human PVR ), amino acids No 419 - 590 from SEO ID NO : 15 ( human LIRB5 ) , amino 229 -325 from SEQ ID NO : 36 (human CD200 recep acids No 420 -598 from SEQ ID NO :16 (human tori ), amino acids No 181 -386 from SEQ ID NO :33 LIRB2 ) , amino acids No 375 -515 from SEO ID NO : 18 (human TR10D ), amino acids No 230 -468 from SEO ( human FCRL4 ), amino acids No 753 - 977 from SEQ ID NO : 34 (human TR10A ) or amino acids No 179 -440 ID NO : 23 (human FCRL5) , amino acids No 388 -508 from SEQ ID NO : 35 ( human TR10B ) ; from SEQ ID NO : 24 (human FCRL2 ), amino acids No [0851 ] and a CD19 specific chimeric antigen receptor 292 -429 from SEQ ID NO : 25 (human FCRL1) , amino (CAR ) (as P -CAR ) which comprises in the engineered acids No 564 - 734 from SEQ ID NO : 27 ( human immune cell in combination with the N -CAR ; said CD19 US 2017 /0296623 A1 Oct . 19 , 2017 47 specific chimeric antigen receptor (CAR ) having one of the from SEQ ID NO :30 (human PVR ), amino acids No polypeptide structure selected from V1 to V6 , preferably V1, 229 - 325 from SEQ ID NO :36 (human CD200 recep V3 and V5 as illustrated in FIG . 1 , said structure comprising torl ) , amino acids No 181 -386 from SEQ ID NO :33 an extra cellular ligand binding - domain comprising VH and (human TR10D ) , amino acids No 230 -468 from SEQ VL from a monoclonal anti -CD19 antibody, a hinge , a ID NO : 34 (human TR10A ) or amino acids No 179 - 440 transmembrane domain and a cytoplasmic domain including from SEQ ID NO :35 (human TR10B ); a CD3 zeta signaling domain and a co - stimulatory domain [0861 ] and a CD19 specific chimeric antigen receptor from 4 - 1BB . (CAR ) ( as P - CAR ) which comprises in the engineered [0852 ] According to a preferred embodiment, the isolated immune cell in combination with the N -CAR ; said CD19 immune cell includes at least a N -CAR which comprises at specific chimeric antigen receptor (CAR ) containing at least least : one transmembrane polypeptide which includes at least one [0853 ] an extracellular binding domain , which is able to extra -binding domain recognizing specifically the CD19 bind to an “ off - target" antigen on a healthy cell ; antigen , and an intracellular signaling domain , optionally [0854 ] and ; with co - stimulatory domain . [ 0855 ] an inhibitory transmembrane polypeptide having a polypeptide sequence of more than 80 % , preferably [0862 ] According to a preferred embodiment, the isolated 90 % and more preferably 95 % identity with SEQ ID immune cell includes at least a N -CAR which comprises at NO : 33 (human TR10D ), SEQ ID NO : 34 (human least: TR10A ) and SEQ ID NO : 35 (human TR10B ); [0863 ] an extracellular binding domain , which is able to [ 0856 ] and a CD19 specific chimeric antigen receptor bind to an “ off- target " antigen on a healthy cell; (CAR ) ( as P - CAR ) which comprises in the engineered [ 0864 ] and ; immune cell in combination with the N -CAR ; said CD19 [0865 ] an inhibitory transmembrane polypeptide having specific chimeric antigen receptor (CAR ) having one of the a sequence consisting essentially of amino acids No polypeptide structure selected from V1 to V6 , preferably V1 , 201 - 375 from SEQ ID NO : 8 (human KIZLA ) , amino V3 and V5 as illustrated in FIG . 1 , said structure comprising acids No 206 - 348 from SEQ ID NO : 2 (human an extra cellular ligand binding -domain comprising VH and KIR2DL1 ), amino acids No 206 - 348 from SEQ ID VL from a monoclonal anti -CD38 antibody, a hinge , a NO : 1 ( human KIR2DL2) , amino acids No 206 - 341 transmembrane domain and a cytoplasmic domain including from SEQ ID NO : 4 ( human KIR2DL3 ) , amino acids a CD3 zeta signaling domain and a co - stimulatory domain No 206 - 348 from SEQ ID NO : 2 (human KIR2DL1) , from 4 - 1BB . amino acids No 203 -377 from SEQ ID NO : 6 (human [ 0857 ] According to an embodiment, the isolated immune KIR2DL4 ) , amino acids No 301- 444 from SEQ ID cell includes at least a N -CAR which comprises at least : NO : 7 (human KIR3DL1) , amino acids No 301- 455 [ 0858 ] an extracellular binding domain , which is able to from SEQ ID NO :5 (human KIR3DL2 ), amino acids bind to an “ off- target " antigen on a healthy cell; No 204 - 310 from SEQ ID NO : 24 (human FRGR2B ) , [0859 ] and ; amino acids No 214 - 343 from SEQ ID NO : 9 (human [0860 ] an inhibitory transmembrane polypeptide having MILR1) , amino acids No 216 -448 from SEQ ID NO : 10 a sequence consisting essentially of amino acids No (human LIRB4 ), amino acids No 420 -631 from SEQ 201- 375 from SEQ ID NO : 8 (human KIZLA ), amino ID NO :11 (human LIRB3) , amino acids No 296 -410 acids No 206 - 348 from SEO ID NO : 2 (human from SEQ ID NO : 12 (human KI3L3 ) , amino acids No KIR2DL1) , amino acids No 206 -348 from SEQ ID 419 -590 from SEQ ID NO : 15 (human LIRB5) , amino NO : 1 (human KIR2DL2 ) , amino acids No 206 - 341 acids No 420 - 598 from SEQ ID NO : 16 (human from SEQ ID NO : 4 (human KIR2DL3 ) , amino acids LIRB2 ) , amino acids No 375 -515 from SEO ID NO : 18 No 206 -348 from SEQ ID NO : 2 ( human KIR2DL1) , (human FCRL4 ), amino acids No 753 - 977 from SEQ amino acids No 203 - 377 from SEQ ID NO :6 (human ID NO :23 (human FCRL5) , amino acids No 388 - 508 KIR2DL4 ) , amino acids No 301- 444 from SEQ ID from SEO ID NO : 24 ( human FCRL2 ) , amino acids No NO : 7 (human KIR3DL1) , amino acids No 301- 455 292 - 429 from SEQ ID NO : 25 (human FCRL1 ) , amino from SEO ID NO : 5 (human KIR3DL2 ) , amino acids acids No 564 - 734 from SEQ ID NO : 27 ( human No 204 -310 from SEQ ID NO :24 (human FRGR2B ) , FCRL3 ), amino acids No 147 -269 from SEQ ID NO :28 amino acids No 214 - 343 from SEQ ID NO : 9 ( human (human MPZL1) , amino acids No 151 - 303 from SEQ MILR1) , amino acids No 216 - 448 from SEQ ID NO : 10 ID NO : 29 (human PILRA ) , amino acids No 329 -417 (human LIRB4 ) , amino acids No 420 - 631 from SEO from SEQ ID NO :30 (human PVR ), amino acids No ID NO : 11 ( human LIRB3 ) , amino acids No 296 - 410 229 -325 from SEQ ID NO : 36 ( human CD 200 recep from SEQ ID NO : 12 (human KI3L3 ), amino acids No torl ) , amino acids No 181 -386 from SEQ ID NO :33 419 -590 from SEQ ID NO : 15 (human LIRB5 ), amino (human TR10D ) , amino acids No 230 -468 from SEQ acids No 420 - 598 from SEQ ID NO : 16 human ID NO : 34 ( human TR10A ) or amino acids No 179 -440 LIRB2) , amino acids No 375 -515 from SEQ ID NO : 18 from SEQ ID NO :35 ( human TR10B ) ; (human FCRL4 ) , amino acids No 753 - 977 from SEO [0866 ] and a CD19 specific chimeric antigen receptor ID NO :23 (human FCRL5 ) , amino acids No 388 - 508 (CAR ) ( as P -CAR ) which comprises in the engineered from SEQ ID NO : 24 (human FCRL2) , amino acids No immune cell in combination with the N -CAR ; said CD19 292 - 429 from SEQ ID NO : 25 ( human FCRL1) , amino specific chimeric antigen receptor (CAR ) having one of the acids No 564 -734 from SEQ ID NO :27 (human polypeptide structure selected from V1 to V6 , preferably V1, FCRL3 ) , amino acids No 147 - 269 from SEQ ID NO :28 V3 and V5 as illustrated in FIG . 1 , said structure comprising (human MPZL1) , amino acids No 151 - 303 from SEQ an extra cellular ligand binding - domain comprising VH and ID NO : 29 (human PILRA ) , amino acids No 329 -417 VL from a monoclonal anti -CD19 antibody, a hinge , a US 2017 /0296623 A1 Oct . 19 , 2017 48 transmembrane domain and a cytoplasmic domain including NO : 33 (human TR10D ), SEQ ID NO : 34 (human a CD3 zeta signaling domain and a co - stimulatory domain TR10A ) and SEO ID NO : 35 ( human TR10B ) ; from 4 - 1BB . f0876 ] and a CD19 specific chimeric antigen receptor [0867 ] According to a preferred embodiment , the isolated ( CAR ) (as P -CAR ) which comprises in the engineered immune cell includes at least a N - CAR which comprises at immune cell in combination with the N -CAR ; said CD19 least : specific chimeric antigen receptor (CAR ) having one of the [0868 ] an extracellular binding domain , which is able to polypeptide structure selected from V1 to V6 , preferably V1 , bind to an “ off - target ” antigen on a healthy cell ; V3 and V5 as illustrated in FIG . 1 , said structure comprising [ 0869] and ; an extra cellular ligand binding -domain comprising VH and [ 0870 ] an inhibitory transmembrane polypeptide having VL from a monoclonal anti -CD19 antibody , a hinge , a a sequence consisting essentially of amino acids No transmembrane domain and a cytoplasmic domain including 201 - 375 from SEO ID NO : 8 (human KIZLA ) , amino a CD3 zeta signaling domain and a co - stimulatory domain acids No 206 - 348 from SEQ ID NO : 2 (human from 4 - 1BB . KIR2DL1) , amino acids No 206 - 348 from SEQ ID [0877 ] Said immune cell engineered to express both the NO : 1 (human KIR2DL2 ) , amino acids No 206 - 341 N -CAR and the P -CAR such as presented previously is from SEQ ID NO : 4 (human KIR2DL3) , amino acids intended to be used as a medicament. No 206 - 348 from SEQ ID NO : 2 (human KIR2DL1) , [0878 ] Preferably, such engineered immune cell is amino acids No 203 -377 from SEQ ID NO :6 (human intended to be used for the treatment of cancer . KIR2DL4 ) , amino acids No 301 - 444 from SEQ ID [0879 ] More preferably , such engineered immune cell is NO : 7 (human KIR3DL1) , amino acids No 301- 455 intended to be used for the treatment of refractory relapsed from SEQ ID NO :5 (human KIR3DL2 ), amino acids cancer . No 204 -310 from SEQ ID NO : 24 ( human FRGR2B ), (0880 ] By " relapsed cancer " , it is referred to a cancer that amino acids No 214 -343 from SEQ ID NO : 9 (human returns after a period of improvement. This applies whether MILR1 ) , amino acids No 216 - 448 from SEO ID NO : 10 the cancer was treated or untreated . ( human LIRB4 ) , amino acids No 420 -631 from SEO 0881 ] By “ refractory cancer ” , it is referred to a cancer ID NO : 11 (human LIRB3 ) , amino acids No 296 -410 that proves resistant, or does not respond to , treatment, from SEQ ID NO : 12 (human KI3L3 ), amino acids No regardless whether the cancer is resistant to treatment imme 419 -590 from SEO ID NO : 15 ( human LIRB5 ) , amino diately , or it develops a resistance during treatment. acids No 420 - 598 from SEQ ID NO : 16 (human [0882 ] Methods of Engineering Immune Cells LIRB2) , amino acids No 375 - 515 from SEQ ID NO : 18 [0883 ] The inventors developed methods of engineering (human FCRL4 ) , amino acids No 753 - 977 from SEQ such immune cells based on the rational combination of ID NO : 23 (human FCRL5 ) , amino acids No 388 - 508 regulatory modules in artificial circuits for performing tasks from SEQ ID NO :24 (human FCRL2 ), amino acids No based on “ NOT gates” . The term “ gate” is used to refer to 292 - 429 from SEO ID NO : 25 (human FCRL1) , amino a device or molecular mechanism that produces a particular acids No 564 - 734 from SEQ ID NO : 27 (human ( predetermined ) output in response to two or more input FCRL3) , amino acids No 147 - 269 from SEQ ID NO :28 signals . According to the present invention , the logical NOT ( human MPZL1) , amino acids No 151 - 303 from SEQ gate refers to the immune cell inhibition , in particular T cell ID NO :29 (human PILRA ) , amino acids No 329 - 417 cytotoxicity against a target cell through inhibition of spe from SEQ ID NO :30 (human PVR ), amino acids No cific proteins (signaling proteins) resulting from the con 229 - 325 from SEQ ID NO : 36 (human CD200 recep comitant binding to 2 different antigens . tori ) , amino acids No 181 - 386 from SEQ ID NO : 33 [ 0884 ] In one embodiment, the method of engineering an ( human TR10D ), amino acids No 230 - 468 from SEQ immune can comprise the steps of: ID NO : 34 ( human TR10A ) or amino acids No 179 - 440 [0885 ] ( a ) Providing an immune cell ; from SEQ ID NO : 35 (human TR10B ) ; [0886 ] (b ) Expressing the N -CAR and the P -CAR at the [ 0871 ] and a CD19 specific chimeric antigen receptor surface of said cell . (CAR ) ( as P -CAR ) which comprises in the engineered 108871 In another embodiment, the method of engineering immune cell in combination with the N -CAR ; said CD19 an immune cell can comprise the steps of: specific chimeric antigen receptor (CAR ) having one of the 10888 ) a . Introducing into said cell at least one polynucle polypeptide structure selected from V1 to V6 , preferably V1, otide encoding the N - CAR and at least one polynucleotide V3 and V5 as illustrated in FIG . 1 , said structure comprising encoding the CAR ; an extra cellular ligand binding -domain comprising VH and [0889 ] b . Expressing said polynucleotides into said cell . VL from a monoclonal anti -CD19 antibody, a hinge , a [0890 ] P -CARs and immune cells comprising them have transmembrane domain and a cytoplasmic domain including been extensively disclosed and can be prepared by the a CD3 zeta signaling domain and a co - stimulatory domain skilled person according to known methods. For example , a from 4 - 1BB . methodology to prepare P -CAR and cells comprising such [ 0872] According to a preferred embodiment, the isolated P -CARs is disclosed in U . S . Pat. No . 7 ,446 , 190 , WO2008 / immune cell includes at least a N -CAR which comprises at 121420 , U . S . Pat. No . 8 ,252 , 592 , US20140024809, least : WO2012 / 079000 , WO2014153270 , WO2012 /099973 , [0873 ] an extracellular binding domain , which is able to WO2014 /011988 , WO2014 /011987 , WO2013 / 067492 , bind to an “ off -target ” antigen on a healthy cell; WO2013 /070468 , WO2013 /040557 , WO2013 / 126712 , [0874 ] and ; WO2013 / 126729 , WO 2013 / 126726 , WO2013 / 126733 , [ 0875 ] an inhibitory transmembrane polypeptide having U . S . Pat. No . 8 , 399 ,645 , US20130266551, a polypeptide sequence of more than 80 % , preferably US20140023674 , WO2014039523 , U . S . Pat . No . 7 ,514 ,537 , 90 % and more preferably 95 % identity with SEQ ID U . S . Pat. No. 8 ,324 ,353 , WO2010 /025177 , U . S . Pat. No. US 2017 /0296623 A1 Oct . 19, 2017 49 7 , 446 , 179 , WO2010 / 025177 , WO2012 /031744 , WO2012 / [0914 ] An immunosuppressive agent is an agent that sup 136231A1, WO2012 / 050374A2 , WO2013074916 , presses immune function by one of severalmechanisms of WO2009 /091826A3 , WO2013 / 176915 or WO / 2013 /059593 action . In other words, an immunosuppressive agent is a role which are all incorporated herein in their entirety by refer played by a compound which is exhibited by a capability to ence . Immune cells comprising a P -CAR and a N -CAR can diminish the extent and /or voracity of an immune response . be prepared by the skilled person according to the method As non - limiting example , an immunosuppressive agent can ologies disclosed in the above mentioned references . In a be a calcineurin inhibitor, a target of rapamycin , an inter preferred embodiment, immune cells comprising a P -CAR leukin - 2 u - chain blocker, an inhibitor of inosine monophos and a N -CAR can be prepared by the skilled person accord phate dehydrogenase , an inhibitor of dihydrofolic acid ing to the methodology disclosed in WO2013/ 176915 . reductase , a corticosteroid or an immunosuppressive anti [0891 ] In one embodiment, the method of engineering metabolite . T- cells of invention can comprise : [0915 ] In a particular embodiment, the genetic modifica [ 0892 ] (a ) Providing a T - cell, preferably from a cell tion step of the method relies on the inactivation of one gene culture or from a blood sample ; selected from the group consisting ofCD52 , GR , TCR alpha [0893 ] (b ) Transforming said T cell with a nucleic acid and TCR beta . In another embodiment, the genetic modifi encoding a rare -cutting endonuclease able to cation step of the method relies on the inactivation of two [0894 ] Selectively inactivate by DNA cleavage, preferably genes selected from the group consisting of dCK , CD52 and by double - strand break respectively at least one gene encod GR , CD52 and TCR alpha , CDR52 and TCR beta , GR and ing a component of the T -cell receptor ( TCR ) ; TCR alpha , GR and TCR beta , TCR alpha and TCR beta. In [0895 ] (d ) Expressing said rare -cutting endonucleases another embodiment , the genetic modification step of the into said T - cells ; method relies on the inactivation of more than two genes . [0896 ] ( e ) Sorting the transformed T -cells , which do not The genetic modification is preferably operated ex - vivo . express TCR on their cell surface ; [ 0916 ] Inactivation of CD52 , CTLA - 4 and / or PD - 1 genes , [ 0897] In some embodiments , the method of engineering for instance by TALE -nuclease , may be performed such as T - cells of invention can comprise : described respectively in Examples 2 , 3 and 4 in the appli [0898 ] ( a ) Providing a T -cell , preferably from a cell cation WO2014 / 184744 . culture or from a blood sample ; 0917 ) The rare - cutting endonucleases used for inactivat [0899 ] (b ) Selecting a gene in said T -cell expressing a ing the genes in T -cells are preferably Transcription Acti target for an immunosuppressive agent ; vator like Effector ( TALE ), but may be also a Cas9 coupled [0900 ] ( c ) Transforming said T cell with nucleic acid to a RNA guide as respectively described in WO 2013/ encoding a rare - cutting endonuclease able to selec 176915 and WO 2014 / 191128 . tively inactivate by DNA cleavage, preferably by [ 0918 ] Compositions / Formulations double -strand break respectively : [09191 . Another aspect of the present invention relates to [ 0901 ] said gene encoding a target for said immuno compositions or formulations containing genetically engi suppressive agent, and neered immune cells which express at least one N -CAR and [0902 ) at least one gene encoding a component of the at least one P -CAR such as described above and at least one T -cell receptor ( TCR ) ; pharmaceutically acceptable carrier or vehicle . [0903 ] ( d ) Expressing said rare -cutting endonucleases [0920 ] Compositions of the invention comprising geneti into said T - cells ; cally modified immune cells can be conveniently provided [0904 ] ( e ) Sorting the transformed T -cells , which do not as sterile liquid preparations , e . g ., isotonic aqueous solu express TCR on their cell surface ; tions, suspensions, emulsions, dispersions, or viscous com [0905 ] ( f ) Expanding said cells , optionally in presence positions, which may be buffered to a selected pH . Liquid of said immunosuppressive agent. preparations are normally easier to prepare than gels , other [0906 ] Such inactivation of TCR gene may be performed viscous compositions, and solid compositions. Additionally , such as described in the Example 1 of the application liquid compositions are somewhat more convenient to WO2014 / 184143 . administer, especially by injection . Viscous compositions, [0907 ] In some embodiment , the method to engineer A cell on the other hand , can be formulated within the appropriate of the invention further comprises one or more additional viscosity range to provide longer contact periods with spe genomic modification step . By additional genomic modifi cific tissues . Liquid or viscous compositions can comprise cation step , can be intended the introduction into cells to carriers, which can be a solvent or dispersing medium engineer of one or more protein of interest. Said protein of containing , for example , water , saline , phosphate buffered interest can be a P - CAR and / or an N - CAR . saline, polyol ( for example , glycerol, propylene glycol, [ 0908 ] By “ Immunosuppressive agents ” or “ immunosup liquid polyethylene glycol, and the like ) and suitable mix pressive agents” , it is meant drugs that inhibit or prevent tures thereof. activity of the immune system . [0921 ] Sterile injectable solutions can be prepared by [ 0909 ]. In some embodiments , the method of engineering incorporating the genetically modified immunoresponsive T -cells of invention can comprise : cells utilized in practicing the present invention in the [0910 ] (a ) modifying T -cells by inactivating at least : required amount of the appropriate solvent with various [0911 ] a first gene expressing a target for an immu amounts of the other ingredients, as desired . Such compo nosuppressive agent, and sitions may be in admixture with a suitable carrier, diluent, [0912 ] a second gene encoding a component of the or excipient such as sterile water , physiological saline , T -cell receptor ( TCR ) glucose, dextrose , or the like. The compositions can also be [0913 ] (b ) expanding said cells , optionally in presence lyophilized . The compositions can contain auxiliary sub of said immunosuppressive agent. stances such as wetting , dispersing, or emulsifying agents US 2017 /0296623 A1 Oct . 19 , 2017 50 ( e. g ., methylcellulose ) , pH buffering agents , gelling or vis ably about 0 .0001 to about 0 .05 wt % or about 0 . 001 to about cosity enhancing additives , preservatives , flavoring agents , 20 wt % , preferably about 0 .01 to about 10 wt % , and still colors , and the like , depending upon the route of adminis more preferably about 0 . 05 to about 5 wt % . Of course , for tration and the preparation desired . Standard texts , such as any composition to be administered to an animal or human , “ REMINGTON ' S PHARMACEUTICAL SCIENCE ” , 17th and for any particular method of administration , it is pre edition , 1985 , incorporated herein by reference , may be ferred to determine therefore: toxicity , such as by determin consulted to prepare suitable preparations, without undue ing the lethal dose (LD ) and LD50 in a suitable animal experimentation . model e . g . , rodent such as mouse ; and , the dosage of the [0922 ] Various additives which enhance the stability and composition (s ), concentration of components therein and sterility of the compositions, including antimicrobial preser timing of administering the composition ( s) , which elicit a vatives , antioxidants , chelating agents , and buffers , can be suitable response . Such determinations do not require undue added . Prevention of the action of microorganisms can be experimentation from the knowledge of the skilled artisan , ensured by various antibacterial and antifungal agents , for this disclosure and the documents cited herein . And , the time example , parabens , chlorobutanol, phenol, sorbic acid , and for sequential administrations can be ascertained without the like . Prolonged absorption of the injectable pharmaceu undue experimentation . tical form can be brought about by the use of agents delaying [0927 ] Delivery Methods absorption , for example , aluminium monostearate and gela [0928 ] The different methods described above involve tin . According to the present invention , however, any expressing N - CAR and P -CAR at the surface of a cell . As vehicle , diluent, or additive used would have to be compat non - limiting example , said N -CAR and P -CAR can be ible with the genetically modified immunoresponsive cells expressed by introducing the latter into a cell. CARs can be or their progenitors . introduced as transgene encoded by one plasmidic vector. [ 0923] The compositions can be isotonic , i. e . , they can Said plasmid vector can also contain a selection marker have the same osmotic pressure as blood and lacrimal fluid . which provides for identification and / or selection of cells The desired isotonicity of the compositions of this invention which received said vector. may be accomplished using sodium chloride , or other phar [ 0929 ] Polypeptides may be synthesized in situ in the cell maceutically acceptable agents such as dextrose , boric acid , as a result of the introduction of polynucleotides encoding sodium tartrate , propylene glycol or other inorganic or said polypeptides into the cell . Alternatively , said polypep organic solutes . Sodium chloride is preferred particularly for tides could be produced outside the cell and then introduced buffers containing sodium ions. thereto . Methods for introducing a polynucleotide construct [0924 ] Viscosity of the compositions, if desired , can be into cells are known in the art and including as non - limiting maintained at the selected level using a pharmaceutically examples stable transformation methods wherein the poly acceptable thickening agent. Methylcellulose is preferred nucleotide construct is integrated into the genome of the cell , because it is readily and economically available and is easy transient transformation methods wherein the polynucle to work with . Other suitable thickening agents include , for otide construct is not integrated into the genome of the cell example , xanthan gum , carboxymethyl cellulose , hydroxy and virus mediated methods. Said polynucleotides may be propyl cellulose , carbomer, and the like . The preferred introduced into a cell by for example , recombinant viral concentration of the thickener will depend upon the agent vectors ( e . g . retroviruses , adenoviruses ), liposome and the selected . The important point is to use an amount that will like . For example , transient transformation methods include achieve the selected viscosity . Obviously , the choice of for example microinjection , electroporation or particle bom suitable carriers and other additives will depend on the exact bardment. Said polynucleotides may be included in vectors, route of administration and the nature of the particular more particularly plasmids or virus, in view of being dosage form , e . g ., liquid dosage form ( e . g . , whether the expressed in cells . composition is to be formulated into a solution , a suspen [0930 ] Polynucleotides and Vectors sion , gel or another liquid form , such as a time release form 10931] In one embodiment, said isolated cell according to or liquid - filled form ) . the present invention comprises a polynucleotide encoding [0925 ] Those skilled in the art will recognize that the the “ NOT gate ” receptor (N -CAR & P -CAR ). components of the compositions should be selected to be [0932 ] The present invention also relates to polynucle chemically inert and will not affect the viability or efficacy otides , vectors encoding the above described N -CAR and of the genetically modified immunoresponsive cells as P -CAR according to the invention . described in the present invention . This will present no [0933 ] The polynucleotide may consist in an expression problem to those skilled in chemical and pharmaceutical cassette or expression vector ( e . g . a plasmid for introduction principles , or problems can be readily avoided by reference into a bacterial host cell, or a viral vector such as a to standard texts or by simple experiments (not involving baculovirus vector for transfection of an insect host cell , or undue experimentation ) , from this disclosure and the docu a plasmid or viral vector such as a lentivirus for transfection ments cited herein . of a mammalian host cell ) . [0926 ] The skilled artisan can readily determine the 0934 ] In a particular embodiment, the different nucleic amount of cells and optional additives , vehicles , and/ or acid sequences can be included in one polynucleotide or carrier in compositions and to be administered in methods of vector which comprises a nucleic acid sequence encoding the invention . Typically , any additives in addition to the ribosomal skip sequence such as a sequence encoding a 2A active cell ( s ) and/ or agent ( s ) ) are present in an amount of peptide . 2A peptides, which were identified in the Aphtho 0 .001 to 50 % (weight ) solution in phosphate buffered saline , virus subgroup of picornaviruses , causes a ribosomal “ skip ” and the active ingredient is present in the order of micro from one codon to the next without the formation of a grams to milligrams, such as about 0 .0001 to about 5 wt % , peptide bond between the two amino acids encoded by the preferably about 0 .0001 to about 1 wt % , still more prefer codons ( see (Doronina , Wu et al. 2008, Mol Cell Biol US 2017 /0296623 A1 Oct . 19, 2017 28 ( 13 ) :4227 - 39 ) . By " codon " is meant three nucleotides on by CD123 -expressing cells . In particular, such condition is an mRNA ( or on the sense strand of a DNA molecule ) that characterized by an overabundance of CD123 - expressing are translated by a ribosome into one amino acid residue . cells . Thus , two polypeptides can be synthesized from a single , [ 0944 ] In a particular embodiment, such engineered cell is contiguous open reading frame within an mRNA when the intended for its use in therapy , wherein the leukemia is acute polypeptides are separated by a 2A oligopeptide sequence myelogenous leukemia (AML ) . Therefore, this is particu that is in frame. Such ribosomal skip mechanisms are well larly adapted for treating the pre -malignant or malignant known in the art and are known to be used by several vectors cancer AML condition characterized especially by CD123 for the expression of several proteins encoded by a single expressing cells or by CLL - 1 expressing cells . messenger RNA . [0945 ] In a preferred embodiment, said engineered cell is [0935 ] To direct , transmembrane polypeptide into the intended to be used in therapy, wherein the condition is a secretory pathway of a host cell , a secretory signal sequence pre -malignant or malignant cancer condition such as mul ( also known as a leader sequence , prepro sequence or pre tiple myeloma (MM ) characterized especially by CD38 sequence ) is provided in polynucleotide sequence or vector expressing cells . sequence . The secretory signal sequence is operably linked [0946 ] In another particular embodiment, such engineered to the transmembrane nucleic acid sequence , i. e ., the two cell is intended for its use in therapy, wherein the leukemia sequences are joined in the correct reading frame and is chronic lymphocytic leukemia (CLL ). Therefore , this is positioned to direct the newly synthesized polypeptide into particularly adapted for treating the pre -malignant or malig the secretory pathway of the host cell. Secretory signal nant cancer CLL condition characterized especially by CS1 sequences are commonly positioned 5 ' to the nucleic acid expressing cells. sequence encoding the polypeptide of interest , although [0947 ] In another particular embodiment, said engineered certain secretory signal sequences may be positioned else cell is intended to be used in therapy , wherein the condition where in the nucleic acid sequence of interest ( see , e . g ., is a pre -malignant or malignant cancer CLL condition char Welch et al. , U . S . Pat . No . 5 ,037 ,743 ; Holland et al. , U . S . acterized by ROR1 - expressing cells . Pat. No. 5 , 143, 830 ) . [0948 ] In another embodiment, said engineered cell for [ 0936 ] Those skilled in the art will recognize that, in view use in therapy , wherein said malignant lymphoproliferative of the degeneracy of the genetic code, considerable sequence disorder is lymphoma . More specifically , such engineered variation is possible among these polynucleotide molecules. cell may be used to treat lymphoma is selected from the Preferably , the nucleic acid sequences of the present inven group consisting of multiple myeloma , non - Hodgkin ' s lym tion are codon -optimized for expression in mammalian cells , phoma, Burkitt ' s lymphoma, and follicular lymphoma preferably for expression in human cells . Codon -optimiza (small cell and large cell) . tion refers to the exchange in a sequence of interest of [0949 ] In another particular embodiment, said engineered codons that are generally rare in highly expressed genes of cell is intended to be used in therapy , wherein the condition a given species by codons that are generally frequent in is a solid tumor such as breast, colon , lung , or kidney tumor highly expressed genes of such species , such codons encod characterized especially by ROR1 -expressing cells . [0950 ] In a preferred embodiment, said engineered cell is ing the amino acids as the codons that are being exchanged . intended to be used in therapy , wherein the condition is a [ 0937 ] Therapeutic Applications pre -malignant or malignant cancer condition characterized [0938 ] In another embodiment, isolated cell expressing by CD22 - expressing cells . both at least one N - CAR and at least one P -CAR obtained 10951] In another aspect , the present invention relies on by the different methods or cell line derived from said methods for treating patients in need thereof , said method isolated cell as previously described can be used as a comprising at least one of the following steps : medicament. [0952 ] ( a ) providing an immune - cell obtainable by any [0939 ] In another embodiment, said medicament can be one of the methods previously described ; used for treating cancer in a patient in need thereof. [0953 ] ( b ) Administrating said transformed immune cells [ 0940 ] In another embodiment, said isolated cell accord to said patient, ing to the invention or cell line derived from said isolated [0954 ] On one embodiment , said T cells of the invention cell can be used in the manufacture of a medicament for can undergo robust in vivo T cell expansion and can persist treatment of a cancer in a patient in need thereof. for an extended amount of time. [ 0955 ] Said treatment can be ameliorating , curative or [ 0941] In another embodiment , said engineered cell prophylactic . It may be either part of an autologous immu expressing both at least one N -CAR and at least one P -CAR notherapy or part of an allogenic immunotherapy treatment . is intended for its use in therapy, wherein the condition is a By autologous , it is meant that cells , cell line or population haematological cancer condition . In particular, such haema of cells used for treating patients are originating from said tological cancer condition is leukemia . patient or from a Human Leucocyte Antigen ( HLA ) com [ 0942 ] More specifically , such engineered cell is intended patible donor. By allogeneic is meant that the cells or for its use in therapy, wherein said leukemia is selected from population of cells used for treating patients are not origi the group consisting of acute myelogenous leukemia nating from said patient but from a donor . (AML ) , chronic myelogenous leukemia , melodysplastic [ 0956 ) Cells that can be used with the disclosed methods syndrome, acute lymphoid leukemia , chronic lymphoid leu such as TALE nuclease . kemia (CLL ) , and myelodysplastic syndrome. [09571 Said treatment can be used to treat patients diag [ 0943 ] In a preferred embodiment, said engineered cell is nosed with cancer , viral infection , autoimmune disorders or intended to be used in therapy, wherein the condition is a Graft versus Host Disease (GVHD ) . Cancers that may be pre -malignant or malignant cancer condition characterized treated include tumors that are not vascularized , or not yet US 2017 /0296623 A1 Oct . 19 , 2017 substantially vascularized , as well as vascularized tumors . other immunoablative agents such as CAMPATH , anti -CD3 The cancers may comprise non solid tumors ( such as hema antibodies or other antibody therapies, cytoxin , fludaribine , tological tumors , for example , leukemias and lymphomas ) cyclosporin , FK506 , rapamycin , mycoplienolic acid , ste or may comprise solid tumors . Types of cancers to be treated roids, FR901228 , cytokines , and irradiation . These drugs with the N - CAR and P - CAR of the invention include, but inhibit either the calcium dependent phosphatase calcineurin are not limited to, carcinoma, blastoma, and sarcoma, and ( cyclosporine and FK506 ) or inhibit the p7056 kinase that is certain leukemia or lymphoid malignancies , benign and important for growth factor induced signaling ( rapamycin ) malignant tumors , and malignancies e . g ., sarcomas , carci (Henderson , Naya et al. 1991 , Immunology 73 ( 3 ) : 316 - 21 ; nomas, and melanomas. Adult tumors /cancers and pediatric Liu , Albers et al . 1992 , 31 ( 16 ): 3896 - 901 ; Bierer, Hollander tumors / cancers are also included . et al. 1993 , Curr Opin Immunol 5 ( 5 ) : 763- 73 ) . In a further [0958 ] It can be a treatment in combination with one or embodiment, the cell compositions of the present invention more therapies against cancer selected from the group of are administered to a patient in conjunction with ( e . g . , antibodies therapy , chemotherapy , cytokines therapy, den before , simultaneously or following ) bone marrow trans dritic cell therapy , gene therapy, hormone therapy, laser light plantation , T cell ablative therapy using either chemotherapy therapy and radiation therapy. agents such as, fludarabine , external- beam radiation therapy [ 0959 ] The administration of the cells or population of (XRT ) , cyclophosphamide, or antibodies such as OKT3 or cells according to the present invention may be carried out CAMPATH . in any convenient manner , including by aerosol inhalation , [0963 ] In another embodiment, the cell compositions of injection , ingestion , transfusion , implantation or transplan the present invention are administered following B - cell tation . The compositions described herein may be adminis ablative therapy such as agents that react with CD20 , e . g . , tered to a patient subcutaneously , intradermally, intratumor Rituxan . For example , in one embodiment, subjects may ally , intranodally, intramedullary , intramuscularly , by undergo standard treatment with high dose chemotherapy intravenous or intralymphatic injection , or intraperitoneally . followed by peripheral blood stem cell transplantation . In In one embodiment, the cell compositions of the present certain embodiments , following the transplant, subjects invention are preferably administered by intravenous injec receive an infusion of the expanded immune cells of the tion . present invention . In an additional embodiment, expanded 0960 ] The administration of the cells or population of cells are administered before or following surgery . cells can consist of the administration of 104 - 101° cells per kg body weight, preferably 10 % to 10º cells /kg body weight Other Definitions including all integer values of cell numbers within those [0964 ] Amino acid residues in a polypeptide sequence ranges. The cells or population of cells can be administrated are designated herein according to the one - letter code, in one or more doses . In another embodiment, said effective in which , for example , Q means Gln or Glutamine amount of cells are administrated as a single dose . In another residue, R means Arg or Arginine residue and D means embodiment, said effective amount of cells are adminis Asp or Aspartic acid residue . trated as more than one dose over a period time. Timing of [0965 ] Nucleotides are designated as follows: one - letter administration is within the judgment of managing physi code is used for designating the base of a nucleoside : a cian and depends on the clinical condition of the patient. The is adenine , t is thymine , c is cytosine, and g is guanine . cells or population of cells may be obtained from any source , For the degenerated nucleotides, r represents g or a such as a blood bank or a donor. While individual needs (purine nucleotides) , k represents g or t , s represents g vary , determination of optimal ranges of effective amounts or c , w represents a ort, m represents a orc, y of a given cell type for a particular disease or conditions represents t or c ( pyrimidine nucleotides ) , d represents within the skill of the art . An effective amount means an g , a or t, v represents g , a or c , b represents g , t or c , h amount which provides a therapeutic or prophylactic benefit . represents a , t or c, and n represents g , a , t or c . The dosage administrated will be dependent upon the age , [0966 ] “ As used herein , “ nucleic acid ” or “ polynucle health and weight of the recipient, kind of concurrent otides ” refers to nucleotides and / or polynucleotides , treatment, if any , frequency of treatment and the nature of such as deoxyribonucleic acid (DNA ) or ribonucleic the effect desired . acid (RNA ) , oligonucleotides , fragments generated by [0961 ] In another embodiment, said effective amount of the polymerase chain reaction (PCR ) , and fragments cells or composition comprising those cells are adminis generated by any of ligation , scission , endonuclease trated parenterally. Said administration can be an intrave action , and exonuclease action . Nucleic acid molecules nous administration . Said administration can be directly can be composed of monomers that are naturally done by injection within a tumor. occurring nucleotides (such as DNA and RNA ), or [0962 ] In certain embodiments of the present invention , analogs of naturally - occurring nucleotides ( e . g . , cells are administered to a patient in conjunction with ( e . g ., enantiomeric forms of naturally -occurring nucleo before , simultaneously or following ) any number of relevant tides) , or a combination of both . Modified nucleotides treatment modalities , including but not limited to treatment can have alterations in sugar moieties and /or in pyrimi with agents such as antiviral therapy, cidofovir and inter dine or purine base moieties . Sugar modifications leukin - 2 , Cytarabine (also known as ARA - C ) or natalizumab include , for example , replacement of one or more treatment for MS patients or efaliztimab treatment for pso hydroxyl groups with halogens , alkyl groups , amines , riasis patients or other treatments for PML patients . In and azido groups, or sugars can be functionalized as further embodiments , the T cells of the invention may be ethers or esters . Moreover, the entire sugar moiety can used in combination with chemotherapy , radiation , immu be replaced with sterically and electronically similar nosuppressive agents , such as cyclosporin , azathioprine , structures, such as aza - sugars and carbocyclic sugar methotrexate , mycophenolate , and FK506 , antibodies, or analogs. Examples of modifications in a base moiety US 2017 /0296623 A1 Oct . 19 , 2017 53 include alkylated purines and pyrimidines , acylated BLV group , lentivirus, spumavirus (Coffin , J . M ., Retroviri purines or pyrimidines , or other well -known heterocy dae : The viruses and their replication , In Fundamental clic substitutes . Nucleic acid monomers can be linked Virology , Third Edition , B . N . Fields, et al ., Eds. , Lippincott by phosphodiester bonds or analogs of such linkages . Raven Publishers , Philadelphia , 1996 ) . Nucleic acids can be either single stranded or double [0971 ] By “ lentiviral vector ” is meant HIV - Based len stranded . tiviral vectors that are very promising for gene delivery [0967 ] By chimeric antigen receptor (CAR ) is intended because of their relatively large packaging capacity , molecules that combine a binding domain against a reduced immunogenicity and their ability to stably component present on the target cell, for example an transduce with high efficiency a large range of different antibody -based specificity for a desired antigen ( e. g ., cell types. Lentiviral vectors are usually generated tumor antigen ) with a T cell receptor -activating intra following transient transfection of three (packaging , cellular domain to generate a chimeric protein that envelope and transfer ) or more plasmids into producer exhibits a specific anti- target cellular immune activity . cells . Like HIV , lentiviral vectors enter the target cell Generally , CAR consists of an extracellular single through the interaction of viral surface glycoproteins chain antibody ( scFv ) fused to the intracellular signal with receptors on the cell surface . On entry, the viral ing domain of the T cell antigen receptor complex zeta RNA undergoes reverse transcription , which is medi chain ( scFv : C ) and have the ability , when expressed in ated by the viral reverse transcriptase complex . The T cells , to redirect antigen recognition based on the product of reverse transcription is a double -stranded monoclonal antibody ' s specificity. linear viral DNA , which is the substrate for viral [0968 ] By “ delivery vector ” or “ delivery vectors” is integration in the DNA of infected cells . By “ integra intended any delivery vector which can be used in the tive lentiviral vectors (or LV ) ” , is meant such vectors as present invention to put into cell contact ( i . e " contact nonlimiting example , that are able to integrate the ing ” ) or deliver inside cells or subcellular compart genome of a target cell . At the opposite by " non ments (i . e “ introducing ” ) agents /chemicals and mol integrative lentiviral vectors ( or NILV ) ” is meant effi ecules (proteins or nucleic acids) needed in the present cient gene delivery vectors that do not integrate the invention . It includes , but is not limited to liposomal genome of a target cell through the action of the virus delivery vectors , viral delivery vectors , drug delivery integrase . vectors , chemical carriers , polymeric carriers , lipo [0972 ] Delivery vectors and vectors can be associated plexes, polyplexes, dendrimers , microbubbles (ultra or combined with any cellular permeabilization tech sound contrast agents ) , nanoparticles , emulsions or niques such as sonoporation or electroporation or other appropriate transfer vectors . These delivery vec derivatives of these techniques. tors allow delivery of molecules, chemicals, macromol [0973 ] by " mutation ” is intended the substitution , dele ecules (genes , proteins ) , or other vectors such as plas tion , insertion of up to one , two , three , four, five , six , mids, peptides developed by Diatos. In these cases , seven , eight, nine , ten , eleven , twelve , thirteen , four delivery vectors are molecule carriers . By " delivery teen , fifteen , twenty , twenty five , thirty , fourty , fifty , or vector” or “ delivery vectors ” is also intended delivery more nucleotides /amino acids in a polynucleotide methods to perform transfection . (cDNA , gene ) or a polypeptide sequence . The mutation [0969 ] The terms " vector ” or “ vectors” refer to a can affect the coding sequence of a gene or its regula nucleic acid molecule capable of transporting another tory sequence . It may also affect the structure of the nucleic acid to which it has been linked . A “ vector” in genomic sequence or the structure / stability of the the present invention includes, but is not limited to , a encoded mRNA . viral vector, a plasmid , a RNA vector or a linear or [ 09741 by " functional variant” is intended a catalyti circular DNA or RNA molecule which may consists of cally active mutant of a protein or a protein domain ; a chromosomal, non chromosomal, semi- synthetic or such mutantmay have the same activity compared to its synthetic nucleic acids. Preferred vectors are those parent protein or protein domain or additional proper capable of autonomous replication ( episomal vector ) ties , or higher or lower activity . and /or expression of nucleic acids to which they are [ 0975 ] " identity ” refers to sequence identity between linked ( expression vectors ). Large numbers of suitable two nucleic acid molecules or polypeptides . Identity vectors are known to those of skill in the art and can be determined by comparing a position in each commercially available . sequence which may be aligned for purposes of com [ 0970 ] Viral vectors include retrovirus, adenovirus, par parison . When a position in the compared sequence is vovirus ( e . g . adenoassociated viruses ) , coronavirus, nega occupied by the same base , then the molecules are tive strand RNA viruses such as orthomyxovirus (e . g ., identical at that position . A degree of similarity or influenza virus ) , rhabdovirus ( e . g ., rabies and vesicular identity between nucleic acid or amino acid sequences stomatitis virus ) , paramyxovirus ( e . g . measles and Sendai) , is a function of the number of identical or matching positive strand RNA viruses such as picornavirus and nucleotides at positions shared by the nucleic acid alphavirus , and double - stranded DNA viruses including sequences. Various alignment algorithms and / or pro adenovirus, herpesvirus ( e . g ., Herpes Simplex virus types 1 grams may be used to calculate the identity between and 2 , Epstein -Barr virus, cytomegalovirus ), and poxvirus two sequences , including FASTA , or BLAST which are ( e . g ., vaccinia , fowlpox and canarypox ) . Other viruses available as a part of the GCG sequence analysis include Norwalk virus, togavirus , flavivirus , reoviruses, package (University of Wisconsin , Madison , Wis .) , and papovavirus , hepadnavirus, and hepatitis virus, for example . can be used with , e .g ., default setting . For example , Examples of retroviruses include: avian leukosis - sarcoma, polypeptides having at least 70 % , 85 % , 90 % , 95 % , mammalian C - type, B - type viruses, D type viruses, HTLV 98 % or 99 % identity to specific polypeptides described US 2017 /0296623 A1 Oct . 19, 2017 54 herein and preferably exhibiting substantially the same vated in X - VivoTM - 15 medium (Lonza ) supplemented with functions, as well as polynucleotide encoding such 20 ng/ mL Human IL - 2 , 5 % Human , and Dynabeads Human polypeptides, are contemplated . T activator CD3/ CD28 at a bead :cell ratio 1 : 1 (Life Tech [0976 ] The term “ subject” or “ patient" as used herein nologies ). includes all members of the animal kingdom including [ 0982 ] CAR mRNA Transfection non - human primates and humans . [0983 ] Transfections were done at Day 4 or Day 11 after [0977 ] By “ Transcription Activator like Effector T -cell purification and activation . 5 millions of cells were ( TALE ) " it is meant a binding domain protein wherein transfected with 15 ug ofmRNA encoding the different CAR sequence specificity is driven by a series of 33 - 35 constructs . CAR mRNAs were produced using T7 mRNA amino acids repeats originating from Xanthomonas or polymerase transfections done using Cytopulse technology , Ralstonia bacterial proteins. These repeats differ essen tially by two amino acids positions that specify an by applying two 0 . 1 ms pulses at 3000V / cm followed by interaction with a base pair (Boch , Scholze et al . 2009 , four 0 . 2 mS pulses at 325V / cm in 0 . 4 cm gap cuvettes in a Science 326 (5959 ) : 1509 - 12 ; Moscou and Bogdanove final volume of 200 ul of “ Cytoporation buffer T” (BTX 2009, Science 326 (5959 ) : 1501 ). Each base pair in the Harvard Apparatus) . Cells were immediately diluted in DNA target is contacted by a single repeat , with the X - VivoTM - 15 media and incubated at 37° C . with 5 % CO2. specificity resulting from the two variant amino acids IL - 2 was added 2 h after electroporation at 20 ng /mL . of the repeat the so - called repeat variable dipeptide, [0984 ] Degranulation Assay (CD107a Mobilization ) RVD ) . TALE binding domains may further comprise an [0985 ] T - cells were incubated in 96 -well plates ( 40 , 000 N -terminal translocation domain responsible for the cells /well ) , together with an equal amount of cells express requirement of a first thymine base ( To ) of the targeted ing various levels of the P antigen (eg expression CD123 or sequence and a C -terminal domain that containing a CD20 ) and undetectable level of N antigen or together with nuclear localization signals (NLS ) . A TALE nucleic an equal amount of cells expressing various levels of the P acid binding domain generally corresponds to an engi antigen (eg expression CD123 or CD20 ) and detectable level neered core TALE scaffold comprising a plurality of of N antigen as determined by flow cytometry analysis using TALE repeat sequences , each repeat comprising a RVD appropriate control( s ) . Co - cultures were maintained in a specific to each nucleotides base of a TALE recognition site . In the present invention , each TALE repeat final volume of 100 ul of X - VivoTM - 15 medium (Lonza ) for sequence of said core scaffold is made of 30 to 42 6 hours at 37° C . with 5 % CO2. CD107a staining was done amino acids , more preferably 33 or 34 wherein two during cell stimulation , by the addition of a fluorescent critical amino acids ( the so -called repeat variable anti -CD107a antibody at the beginning of the co -culture , dipeptide , RVD ) located at positions 12 and 13 medi together with 1 ug /ml of anti -CD49d , 1 ug /ml of anti -CD28 , ates the recognition of one nucleotide of said TALE and 1x Monensin solution . After the 6 h incubation period , binding site sequence ; equivalent two critical amino cells were stained with a fixable viability dye and fluoro acids can be located at positions other than 12 and 13 chrome- conjugated anti -CD8 and analyzed by flow cytom specially in TALE repeat sequence taller than 33 or 34 etry . The degranulation activity was determined as the % of amino acids long . Preferably , RVDs associated with CD8 + /CD107a + cells , and by determining the mean fluo recognition of the different nucleotides are HD for rescence intensity signal (MFI ) for CD107a staining among recognizing C , NG for recognizing T , NI for recogniz CD8 + cells . Degranulation assays were carried out 24 h after ing A , NN for recognizing G or A . In another embodi mRNA transfection . ment, critical amino acids 12 and 13 can be mutated [0986 ] IFN Gamma Release Assay towards other amino acid residues in order to modulate [0987 ] T - cells were incubated in 96 -well plates ( 40 , 000 their specificity towards nucleotides A , T , C and G and cells /well ) , together with cell lines expressing various levels in particular to enhance this specificity . A TALE nucleic acid binding domain usually comprises between 8 and of the P CAR and /or the N - CAR expressed protein . Co 30 TALE repeat sequences . More preferably, said core cultures were maintained in a final volume of 100 ul of scaffold of the present invention comprises between 8 X - VivoTM - 15 medium (Lonza ) for 24 hours at 37° C . with and 20 TALE repeat sequences ; again more preferably 5 % CO , . After this incubation period the plates were cen 15 TALE repeat sequences . It can also comprise an trifuged at 1500 rpm for 5 minutes and the supernatants were additional single truncated TALE repeat sequence recovered in a new plate . IFN gamma detection in the cell made of 20 amino acids located at the C - terminus of culture supernatants was done by ELISA assay . The IFN said set of TALE repeat sequences , i. e . an additional gamma release assays were carried by starting the cell C -terminal half - TALE repeat sequence . co - cultures 24 h after mRNA transfection . [0978 ] By " primary cell ” or “ primary cells ” are intended [0988 ] Cytotoxicity Assay cells taken directly from living tissue ( i . e . biopsy material) [0989 ] T -cells were incubated in 96 -well plates ( 100 ,000 and established for growth in vitro , that have undergone very cells /well ) , together with 10, 000 target cells ( expressing few population doublings and are therefore more represen CD123 ) and 10 ,000 control (CD123neg ) cells in the same tative of the main functional components and characteristics well. Target and control cells were labelled with fluorescent of tissues from which they are derived from , in comparison intracellular dyes (CFSE or Cell Trace Violet ) before co to continuous tumorigenic or artificially immortalized cell culturing them with CAR + T -cells . The co - cultures were lines . incubated for 4 hours at 37° C . with 5 % CO2. After this [ 0979 ] General Methods incubation period , cells were labelled with a fixable viability [0980 ] Primary T -Cell Cultures dye and analyzed by flow cytometry. Viability of each [0981 ] T cells were purified from Buffy coat samples cellular population (target cells or P CARNCAR neg con using Ficoll gradient density medium . The PBMC layer was trol cells ) was determined and the % of specific cell lysis recovered and T cells were purified using a commercially was calculated . Cytotoxicity assays were carried out 48 h available T - cell enrichment kit . Purified T cells were acti after mRNA transfection . US 2017 /0296623 A1 Oct . 19 , 2017 55 [0990 ] T -Cell Transduction positive Jurkat cells using a model cell line expressing both [0991 ] Transduction of T- cells with recombinant lentiviral the P -CAR and N -CAR target antigens, a model cell line vectors expression the CAR was carried out three days after expressing only the P -CAR antigen and a model cell line T - cell purification /activation . CAR detection at the surface expressing only the N -CAR antigen . of T - cells was done using a recombinant protein consisting [0998 ] The cytotoxicity of P -CAR / N -CAR positive cells , on the fusion of the extracellular domain of the human CAR which are tested versus P -CAR positive cells in presence or expressed protein , together with a murine IgG1 Fc fragment. absence of target cells , is presented in the following Table 3 . Binding of this protein to the CAR molecule was detected The results are expressed as a ratio of CD69 fluorescence with a fluorochrome- conjugated secondary antibody target between the T cells expressing P -CAR /N -CAR or P -CAR in ing the mouse Fc portion of the protein , and analyzed by the presence of target cells and in the absence of target cells . flow cytometry . TABLE 3 EXAMPLES Test of cytotoxicity of T cells encoding a P - CAR combined to diverse Example 1 . Design of Inhibitory Gate Receptors N -CARs versus T cells P -CAR only when they are tested with or without target cells ( based on ratio mean CD69 fluorescence [ 0992] Sequences to construct N -CARs are obtained from intensity + / – target cells ) the Uniprot database and were restricted to human proteins, Name of excluding in addition type II membrane proteins ( N - termi plasmid nus on the cytoplasmic side of the membrane ) . encoding N - CAR [0993 ] N - CAR are designed (such as schematized in FIG . inhibitory ( inhibitory P -CAR & N 2 ) to be composed of an antigen targeting domain ( anti sequence sequence ) P - CAR CAR CD20 VH & VL of SEQ ID NO . 45 -46 , anti- BCMA VH & PCLS27705 iFCRL2 1 . 51 1 . 13 VL of SEO ID NO . 41 - 42 and anti - PSMA VH & VL chain pCLS27706 iFCRL3 1 . 58 1 . 14 of SEQ ID NO . 43 -44 ) fused via a short classical- GS - linker PCLS27707 iFCRL4 1 .62 1 . 15 of SEQ ID NO . 39 or SEQ ID NO . 40 to the membrane PCLS27698 iCD200R 1 . 68 1 . 14 receptor of interest of SEQ ID NO . 1 -36 that included the PCLS27708 iFCRL5 1 . 82 1 . 19 PCLS27699 iTR10A 1 . 76 1 . 01 whole cytoplasmic domain , the transmembrane domain and PCLS27709 ILIRB2 1 .68 1 . 17 the amino acid sequence up to the first annotated extracel pCLS27700 iTR10B 1 .62 0 .97 lular topological domain . In case where the extracellular PCLS27711 ILIRB4 1 . 67 1 . 07 topological domains are not clearly annotated , the fusion PCLS27701 iTRIOD 1. 67 1 . 22 point was determined based on other similar receptors . If the PCLS27712 ILIRB5 1 . 70 1 . 14 pCLS27713 IMILR1 1 . 58 1 . 02 resulting extracellular domain was short, an additional por PCLS27703 iFCG2B 1 . 66 1 . 14 tion of the first annotated extracellular topological domain PCLS27714 IMPZL1 1 . 65 1 . 14 was added . PCLS27704 iFCRL1 1 . 72 1 . 10 [ 0994 ] N - CAR are cloned in a mammalian expression PCLS27715 iPILRA 1 .62 1 . 05 plasmid upstream a 2A cis -acting hydrolase element of SEQ PCLS27716 iPVR 1 .54 1 . 11 PCLS27726 iKI2L2 1 . 40 1 . 19 ID NO . 97 followed by a reporter marker ( e . g . fluorescent PCLS27727 IKI2L3 1 . 49 1 . 22 proteins ) of SEQ ID NO . 98 - 99 . Standard molecular biology PCLS27728 iKI2L4 1 .65 1 .19 technics such as PCR , enzymatic restriction digestion and PCLS27729 iKIZLA 1 .64 1 . 21 ligation are applied to create all construction , leading to SEQ PCLS27730 iKI3L1 1 . 70 1 . 13 ID NO . 100 -212 . PCLS27731 iKI3L2 1 .62 1 . 25 PCLS27732 IKI3L3 1 . 64 1 . 15 [0995 ] The production of lentiviral particles to vectorize PCLS27733 iPECA1 1 . 52 1 . 19 CARs and N - CAR is performed using commercially avail 1 .62 1 . 31 able Lentiviral Packaging Mix (Invitrogen ) following the PCLS27725 iKI2L1 manufacturer protocols or, alternatively , the lentiviral par ticles are obtained directly from commercial manufacturers [0999 ] The results shown in the Table 3 that all the (Vectalys ) . N - CARs present a significant inhibitory effect on the P -CAR ( encoding CD20 scFvs ) . This is reflected by a marked Example 2 . Characterization of N -CARS in reduction of the ratio of mean CD69 fluorescence intensity Immortalized Human T- Cells when the T cells endowing both N - CAR and P - CAR in [0996 ] The P -CAR (expressing CD20 antigen ; transduced presence of target cell , when compared to T cells endowing by SEQ ID NO . 213 ) model cell line is generated by P -CAR only . lentiviral transduction of an immortalized human T -cell line [ 1000 ] This is particularly the case for the N -CARs encod ( Jurkat) . The transduced cells are purified for positive sur ing the TRAIL inhibitory receptors TR10A , TR10B and face CAR + expression using bulk FACS sorting or magnetic TR1OD . separation . The whole bulk CAR + population is then assessed for positive CAR + driven activation (degranula Example 3 . Characterization of N -CARS in tion / cytotoxicity ) , proliferation , and cytokine release . The Primary T - Cells results are presented in FIG . 3 . High , medium and low CAR + expressing sub - population or clonal cells are identi [ 1001] N - CAR constructs allowing attenuation of the fied and isolated P - CAR signal (degranulation / cytokine secretion ) identified [ 0997] The appropriate P -CAR ( or CAR + ) Jurkat cell line according to Example 2 are subcloned in a lentiviral pro or population is then transfected with individual or combi duction plasmid using standard molecular biology , leading nation of DNA plasmid encoding N -CARs such as presented to SEQ ID . 214 to 217 . in Example 1 . The level of activation (degranulation / cy [1002 ] Primary T -cells are transduced sequentially using tokine secretion ) is assessed by FACS in P - CARN -CAR N - CAR lentiviral particles SEQ ID . 215 and SEQ ID NO . US 2017 /0296623 A1 Oct . 19 , 2017 56 217 and P -CAR of SEQ ID . 218 . The N - CAR transduced compared to the antigen CAR - high / antigen N -CAR low T - cells are purified for positive surface expression using target cell in the presence of the TR10D engineered N - CAR . FACS sorting. The P -CAR positive population is then trans Indeed , an increase of the ratio ofthe percentage of live cells duced with N -CAR lentiviral particles . Comparative effects between the two target cell populations ( at the three ratio of of P -CAR /N -CAR and N - CAR engineered primary T -cells is assessed using a engineered target cell line (HEK293 ) that target/ effectors are used : 1 / 1 , 1 / 3 and 1 / 10 , FIG . 4 ) is contained two major populations expressing the target anti measured . gens for the P -CAR and N -CAR ( antigen P - CAR high / [ 1004 ] The data show a dose dependent effect in the antigen N -CAR high and antigen P -CAR high / antigen N -CAR low ). The target cell population is then incubated for reduction induced by N -CAR . 6 hours with the different engineered primary T- cells and the [ 1005 ] Surprisingly , when testing a N -CAR comprising relative proportion of the two target populations ( live cells the inhibitory molecule PD - 1 in the same conditions, an expressing CD19 and PMSA antigens ) is recorded . increase in the ratio of % of target cells antigen P -CAR [ 1003 ] The data clearly indicated that the antigen P -CAR high /antigen N -CAR high and antigen P -CAR - high / antigen high /antigen N -CAR high target cell population is protected N -CAR -low is measured . SEQUENCE LISTING The patent application contains a lengthy “ Sequence Listing " section . A copy of the “ Sequence Listing ” is available in electronic form from the USPTO web site ( http :/ / seqdata . uspto . gov / ? pageRequest = docDetail & DocID = US20170296623A1) . An electronic copy of the “ Sequence Listing ” will also be available from the USPTO upon request and payment of the fee set forth in 37 CFR 1 . 19 (b ) ( 3 ). 1. An inhibitory chimeric antigen receptor (N -CAR ) com cell or a healthy cell also expressing a P antigen , said P prising : antigen being also expressed or over expressed on a can an extracellular domain comprising an antigen binding cerous cell. domain ; 7 . The N -CAR according to claim 1 , wherein said antigen binding domain binds to at least one cell surface antigen N a transmembrane domain ; and selected from CD56 , CD205 , CD83 , CD206 , CD200 , CD36 , an intracellular domain ; troponin C , beta - 1 integrin , CCKBR , GALR1 CUBN , CD4 , wherein said N - CAR comprises a polypeptide sequence CD20 , CD22 , CD25 , MUC1, CD19 , or PSMA . involved in inducing an inhibitory transduction signal, 8 . A vector encoding a N -CAR according to claim 1 . said polypeptide sequence comprises at least one sequence 9 . An immune cell comprising a P -CAR comprising : from a Tumor -necrosis - factor related apoptosis inducing an extracellular domain comprising an antigen binding ligand ( TRAIL ) receptor or at least one sequence from a domain ; CD200 receptor 1 . a transmembrane domain ; and 2 . The N -CAR according to claim 1 , comprising a poly an intracellular domain ; peptide sequence from a Tumor -necrosis - factor related and a N -CAR according to claim 1 . apoptosis inducing ligand ( TRAIL ) receptor. 10 . The immune cell according to claim 9 , wherein at least 3. The N - CAR according to claim 1, comprising at least one gene encoding a TCR alpha or a TCR beta subunit is one polypeptide sequence from a polypeptide sequence inactivated . selected from SEQ ID NO : 33 (human TR10D ) , SEQ ID 11 . The immune cell according to claim 9 wherein at least NO : 34 (human TR10A ) or SEQ ID NO : 35 (human TR10B ) one gene encoding a TCR and a gene encoding a deoxycy or fragments thereof. tidine kinase (dck ) are inactivated . 4 . The N -CAR according to claim 1 , wherein said poly 12 . The immune cell according to claim 11 , wherein the peptide sequence has more than 80 % sequence identity with TCR and dck genes are inactivated by deletion using an a sequence from SEQ ID NO : 33 , SEQ ID NO : 34 or SEQ endonuclease . ID NO : 35 or a fragments thereof. 13 . A method of treating or preventing a haematological 5 . The N -CAR according to claim 1 , comprising at least cancer condition , comprising administering to a subject in one of the following polypeptide sequences: amino acids need thereof a therapeutically effective amount of the 181 -386 from SEQ ID NO : 33 (human TR10D ), amino acids immune cell according to claim 12 . 230 - 468 from SEQ ID NO : 34 (human TR10A ) or of amino 14 . The method according to claim 13 , wherein said acids 179 - 440 from SEQ ID NO : 35 (human TR10B ) , or a haematological cancer is leukemia or myeloma. fragments thereof. 15 . A method of engineering an immune cell according to 6 . The N -CAR according to claim 1 , wherein said antigen claim 9 comprising : binding domain binds to a cell surface antigen N , N being ( a ) Providing an immune cell comprising optionally delet not expressed on a cancerous cell and N being expressed on ing a candidate gene, said candidate gene being TCRA a non -cancerous cell or a healthy cell , said non - cancerous or dCK ; US 2017 /0296623 A1 Oct . 19 , 2017 57 ( b ) Expressing a N -CAR and a P -CAR at the cell surface ; and ( c ) optionally deleting a candidate gene, said candidate gene selected from TCRA , PD1, CTLA4 or dCK . 16 . The method of claim 15 , wherein said immune cells are from a donor. 17 . The immune cell according to claim 9 , wherein the immune cell is primary immune T - cell. 18 . The immune cell according to claim 10 , wherein the TCR alpha or the TCR beta subunit gene is inactivated by deletion using a specific endonuclease . 19 . The method according to claim 13, wherein the haematological cancer is a relapsed refractory haematologi cal cancer. 20 . The method according to claim 14 , wherein the leukemia or myeloma is a relapsed and / or refractory leuke mia or myeloma . * * * * *