DOCSLIB.ORG

Cd1d-Unrestricted NKT Cells Are Endowed with a Hybrid Function Far Superior Than That of Inkt Cells

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Cd1d-Unrestricted NKT Cells Are Endowed with a Hybrid Function Far Superior Than That of Inkt Cells CD1d-unrestricted NKT cells are endowed with a hybrid function far superior than that of iNKT cells Alexander R. Farra, Weisheng Wub, Bongkum Choia, James D. Cavalcolib, and Yasmina Laouara,1 Departments of aMicrobiology and Immunology and bComputational Medicine and Bioinformatics, University of Michigan School of Medicine, Ann Arbor, MI 48109 Edited* by Richard A. Flavell, Howard Hughes Medical Institute, Yale School of Medicine, New Haven, CT, and approved June 27, 2014 (received for review December 17, 2013) Invariant natural killer T (iNKT) cells to date represent the best can be misleading, the criteria that best describes an NKT cell is example of cells known to have a hybrid function, representing the ability to perform with a hybrid function between an NK cell both innate and adaptive immunity. Shared phenotypic similarities and a T cell (2). with NK cells together with a rapid response to a cytokine stimulus Nonetheless, the concept of hybrid function is also an elusive and a productive TCR engagement are the features that underline notion allowing for a gradient of functions. A number of works the hybrid nature of iNKT cells. Using these criteria, we provide refer to an NKT hybrid function as the ability of a T cell with molecular and functional evidence demonstrating that CD1d-inde- phenotypic similarities to NK cells to perform with innate-like pendent (CD1dind) NKT cells, a population of CD1d-unrestricted response. The best example of cells endowed with a hybrid NKT NKT cells, are endowed with a hybrid function far superior to that cell function are thought to be iNKT cells (2). In this study, we of iNKT cells: (i) an extensive shared program with NK cells, (ii) provide molecular and functional evidence demonstrating that a closer Euclidian distance with NK cells, and (iii) the ability to CD1dindNKT cells—a population of MHC-unrestricted T cells— respond to innate stimuli (Poly:IC) with cytotoxic potential in the are endowed with a hybrid function that associates them to the ind same manner as NK cells identify a hybrid feature in CD1d NKT NK cell lineage in a manner far superior to the known link be- cells that truly fulfills the dual function of an NK and a T cell. Our ind tween NK and iNKT cells. An extensive shared program with NK finding that CD1d NKT cells are programmed to act like NK cells cells, a similarity in the gene expression profile with NK cells, in response to innate signals while being capable of adaptive and their ability to respond (like NK cells) not only to cytokine responses is unprecedented, and thus might reemphasize CD1d- signals (IL-12 plus IL-18) but also to innate stimuli [in vivo unrestricted NKT cells as a subset of lymphocytes that could treatment with Poly:IC (Fisher)] with massive production of key affect biological processes of antimicrobial and tumor immunity effector players of the cytotoxic pathway collectively identify in a unique way. ahybridfeatureinCD1dindNKT cells that uniquely fulfills the function of an NK cell and a T cell. granzyme B | CD1d | MHC | NK receptors Results ind INFLAMMATION atural killer T (NKT) cells are increasingly regarded as cells Extensive Phenotypic Similarities Place CD1d NKT Cells Closer to the IMMUNOLOGY AND Nendowed with a hybrid function between an NK cell and a T NK Cell Lineage. Based on the differential expression of CD3 and cell (1, 2). The current classification of NKT cells places them of CD1d tetramer, NK1.1-expressing cells can be divided into − into three categories: type I, type II, and NKT-like cells (1). Type three known cell types: NK cells that are among the CD3 gate, I comprises invariant NKT (iNKT) cells that recognize the gly- iNKT cells that are typically identified by positive staining with colipid α-galactosylceramide (α-GalCer) loaded into the MHC CD1d tetramer, and CD1dindNKTcells that are among those class I molecule, CD1d, and contain an invariant TCR repertoire cells that express NK1.1 similar to NK cells and CD3 similar to of Vα14-Jα18 (3–5). Type II NKT cells are also CD1d dependent conventional T cells but negative for CD1d tetramer (Fig. 1A). but do not respond to α-GalCer in the same way as iNKT cells do To confirm that these cells are distinct from the iNKT cell type, (6, 7). NKT-like cells encompass all other NKT cells and are CD1d we examined this phenotype in CD1d-deficient mice and con- ind independent (CD1d ) (8); they are by far the most heterogeneous clusively demonstrated that these cells are bona fide CD1dindNKT and the least characterized. cells (Fig. 1B). Recent studies have increasingly shown a shared expression of + NK cell-related receptors on other effector cells. CD8 T cells Significance are known to up-regulate NK markers, such as NK1.1, and can even respond quickly like NK cells (9). Other work has described The Immunological Genome Project Consortium has recently NKT cells that express NKp46 (10), a marker selectively asso- identified a gene-expression program that links invariant nat- ciated with conventional NK cells and NK22 cells in the gut (11). γδ ural killer T (iNKT) cells to NK cells. While this is true, our study Moreover, T cells have been shown to express NK markers provides molecular and functional evidence demonstrating and display an innate-like response (12). Collectively, these that CD1d-independent NKT cells are uniquely programmed reports converge to raise the following key questions. What with a hybrid function far superior to that of iNKT cells. Such qualifies as an NKT cell? Do the cells need to express only a unique feature enables this subset of NKT cells to contribute NK1.1 and CD3 to be eligible for NKT nomenclature? With the to adaptive immunity (like T cells) and at the same time act as continuous development of both NK and T-cell fields, the sim- a prominent player in innate defense (like NK cells). plistic definition that NKT cells are subsets of T cells that express the NK1.1 marker is becoming increasingly misleading and Author contributions: Y.L. designed research; A.R.F. and B.C. performed research; A.R.F., even inaccurate. For instance, NK1.1 complex is expressed in W.W., B.C., and J.D.C. analyzed data; and A.R.F. and Y.L. wrote the paper. the BALB/c strain but there are allelic divergences with the The authors declare no conflict of interest. polymorphism leading to the PK136 antibody not reacting to *This Direct Submission article had a prearranged editor. the BALB/c NK.1.1 (NKrp1) complex (13). This definition is 1To whom correspondence should be addressed. Email: [email protected]. also limited in the C57BL/6 strain because of the discovery of − + This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10. NK1.1 CD1d NKT cells (14). Although phenotypic similarities 1073/pnas.1323405111/-/DCSupplemental. www.pnas.org/cgi/doi/10.1073/pnas.1323405111 PNAS | September 2, 2014 | vol. 111 | no. 35 | 12841–12846 Downloaded by guest on September 29, 2021 NK1.1+ NK1.1+ age with a kinetic of development reaching optimum at adult age - A 5 3 B Total cells CD1d-Tatramer ind WT (Fig. S4B). Finally, results from CD1d NKT cell culture in the 2 1.4 11 -/- CD3 CD1d 89 WT presence of γc cytokines suggested that their survival could fit CD1d-Tetramer ** both NK and T-cell lineages (Fig. S5). Taken together, these 2 0.2 15 ind -/- CD1d Frequency (%) results show that CD1d NKT cells, as identified in our study by 95 60 13 + − + + CD3 NK1.1 ind CD3 CD1d tetramer NK1.1 DX5 phenotype, exhibit a unique CD122 CD1d-Tetramer DX5 NK iNKT CD1d NKT DX5 profile at the intersection between NK and T cells. Fig. 1. CD1dindNKT cells resemble NK cells more than iNKT cells resemble + Global Genome Expression Reveals Higher Similarity Between NK and NK cells. (A) Distribution of CD3 versus CD1d tetramer among NK1.1 cells CD1dindNKT Cells than Between NK and iNKT Cells. The Immuno- in the spleen. Distribution of CD122 versus DX5 is shown among gated + + + + + − logical Genome Project Consortium has recently identified NK1.1 CD3 CD1d tetramer (green), NK1.1 CD3 CD1d tetramer (red), + − − and NK1.1 CD3 CD1d tetramer (blue) cells. Numbers indicate the per- a gene-expression program that links iNKT cells to NK cells (2). centage of cells expressing DX5 and high levels of CD122 among each Although it is clear that iNKT cells share patterns with NK cells, ind gate. (B) Distribution of NK1.1 versus CD1d tetramer in the spleen of WT our data placed CD1d NKT cells closer to the NK cell lineage and CD1d−/− mice. (B, Left) Distribution of CD3 versus CD1d tetramer than iNKT cells (Fig. 1 and Figs. S1–S4). To ascertain this + − is from gated NK1.1 CD1d tetramer cells (red). (B, Right) The graph finding, we used a global genome expression analysis and di- − − + + summarizes the frequency of NK (CD3 CD1d tetramer NK1.1 DX5 ), + + + rectly compared the genetic basis for similarities between NK iNKT (CD3 CD1d tetramer ), and CD1dindNKT cells (CD3 CD1d ind − + + − − and CD1d NKT cells versus NK and iNKT cells (Fig. 2 and tetramer NK1.1 DX5 ) in WT and CD1d / mice. Mean ± SEM; **P < 0.001. = Fig. S6). Data are representative of three independent experiments with n 5mice Although iNKT cells confirmed shared patterns with NK cells, per experiment in A and n = 3 mice per experiment in B. our results revealed extensive similarities between CD1dindNKT and NK cells (Fig. 2A). Strikingly, most of these similarities were To better characterize their phenotype, we used a panel of related to the NK cell program, including the killer cell lectin- T- and NK cell markers and directly compared their profile with like receptors (Klr), the NK cell receptors 2B4 (cd244) and that of NK and iNKT cells.
Load more

Recommended publications
  • ENSG Gene Encodes Effector TCR Pathway Costimulation Inhibitory/Exhaustion Synapse/Adhesion Chemokines/Receptors
    ENSG Gene Encodes Effector TCR pathway Costimulation Inhibitory/exhaustion Synapse/adhesion Chemokines/receptors ENSG00000111537 IFNG IFNg x ENSG00000109471 IL2 IL-2 x ENSG00000232810 TNF TNFa x ENSG00000271503 CCL5 CCL5 x x ENSG00000139187 KLRG1 Klrg1 x ENSG00000117560 FASLG Fas ligand x ENSG00000121858 TNFSF10 TRAIL x ENSG00000134545 KLRC1 Klrc1 / NKG2A x ENSG00000213809 KLRK1 Klrk1 / NKG2D x ENSG00000188389 PDCD1 PD-1 x x ENSG00000117281 CD160 CD160 x x ENSG00000134460 IL2RA IL-2 receptor x subunit alpha ENSG00000110324 IL10RA IL-10 receptor x subunit alpha ENSG00000115604 IL18R1 IL-18 receptor 1 x ENSG00000115607 IL18RAP IL-18 receptor x accessory protein ENSG00000081985 IL12RB2 IL-12 receptor x beta 2 ENSG00000186810 CXCR3 CXCR3 x x ENSG00000005844 ITGAL CD11a x ENSG00000160255 ITGB2 CD18; Integrin x x beta-2 ENSG00000156886 ITGAD CD11d x ENSG00000140678 ITGAX; CD11c x x Integrin alpha-X ENSG00000115232 ITGA4 CD49d; Integrin x x alpha-4 ENSG00000169896 ITGAM CD11b; Integrin x x alpha-M ENSG00000138378 STAT4 Stat4 x ENSG00000115415 STAT1 Stat1 x ENSG00000170581 STAT2 Stat2 x ENSG00000126561 STAT5a Stat5a x ENSG00000162434 JAK1 Jak1 x ENSG00000100453 GZMB Granzyme B x ENSG00000145649 GZMA Granzyme A x ENSG00000180644 PRF1 Perforin 1 x ENSG00000115523 GNLY Granulysin x ENSG00000100450 GZMH Granzyme H x ENSG00000113088 GZMK Granzyme K x ENSG00000057657 PRDM1 Blimp-1 x ENSG00000073861 TBX21 T-bet x ENSG00000115738 ID2 ID2 x ENSG00000176083 ZNF683 Hobit x ENSG00000137265 IRF4 Interferon x regulatory factor 4 ENSG00000140968 IRF8 Interferon
    [Show full text]
  • Human and Mouse CD Marker Handbook Human and Mouse CD Marker Key Markers - Human Key Markers - Mouse
    Welcome to More Choice CD Marker Handbook For more information, please visit: Human bdbiosciences.com/eu/go/humancdmarkers Mouse bdbiosciences.com/eu/go/mousecdmarkers Human and Mouse CD Marker Handbook Human and Mouse CD Marker Key Markers - Human Key Markers - Mouse CD3 CD3 CD (cluster of differentiation) molecules are cell surface markers T Cell CD4 CD4 useful for the identification and characterization of leukocytes. The CD CD8 CD8 nomenclature was developed and is maintained through the HLDA (Human Leukocyte Differentiation Antigens) workshop started in 1982. CD45R/B220 CD19 CD19 The goal is to provide standardization of monoclonal antibodies to B Cell CD20 CD22 (B cell activation marker) human antigens across laboratories. To characterize or “workshop” the antibodies, multiple laboratories carry out blind analyses of antibodies. These results independently validate antibody specificity. CD11c CD11c Dendritic Cell CD123 CD123 While the CD nomenclature has been developed for use with human antigens, it is applied to corresponding mouse antigens as well as antigens from other species. However, the mouse and other species NK Cell CD56 CD335 (NKp46) antibodies are not tested by HLDA. Human CD markers were reviewed by the HLDA. New CD markers Stem Cell/ CD34 CD34 were established at the HLDA9 meeting held in Barcelona in 2010. For Precursor hematopoetic stem cell only hematopoetic stem cell only additional information and CD markers please visit www.hcdm.org. Macrophage/ CD14 CD11b/ Mac-1 Monocyte CD33 Ly-71 (F4/80) CD66b Granulocyte CD66b Gr-1/Ly6G Ly6C CD41 CD41 CD61 (Integrin b3) CD61 Platelet CD9 CD62 CD62P (activated platelets) CD235a CD235a Erythrocyte Ter-119 CD146 MECA-32 CD106 CD146 Endothelial Cell CD31 CD62E (activated endothelial cells) Epithelial Cell CD236 CD326 (EPCAM1) For Research Use Only.
    [Show full text]
  • Tools for Cell Therapy and Immunoregulation
    RnDSy-lu-2945 Tools for Cell Therapy and Immunoregulation Target Cell TIM-4 SLAM/CD150 BTNL8 PD-L2/B7-DC B7-H1/PD-L1 (Human) Unknown PD-1 B7-1/CD80 TIM-1 SLAM/CD150 Receptor TIM Family SLAM Family Butyrophilins B7/CD28 Families T Cell Multiple Co-Signaling Molecules Co-stimulatory Co-inhibitory Ig Superfamily Regulate T Cell Activation Target Cell T Cell Target Cell T Cell B7-1/CD80 B7-H1/PD-L1 T cell activation requires two signals: 1) recognition of the antigenic peptide/ B7-1/CD80 B7-2/CD86 CTLA-4 major histocompatibility complex (MHC) by the T cell receptor (TCR) and 2) CD28 antigen-independent co-stimulation induced by interactions between B7-2/CD86 B7-H1/PD-L1 B7-1/CD80 co-signaling molecules expressed on target cells, such as antigen-presenting PD-L2/B7-DC PD-1 ICOS cells (APCs), and their T cell-expressed receptors. Engagement of the TCR in B7-H2/ICOS L 2Ig B7-H3 (Mouse) the absence of this second co-stimulatory signal typically results in T cell B7-H1/PD-L1 B7/CD28 Families 4Ig B7-H3 (Human) anergy or apoptosis. In addition, T cell activation can be negatively regulated Unknown Receptors by co-inhibitory molecules present on APCs. Therefore, integration of the 2Ig B7-H3 Unknown B7-H4 (Mouse) Receptors signals transduced by co-stimulatory and co-inhibitory molecules following TCR B7-H5 4Ig B7-H3 engagement directs the outcome and magnitude of a T cell response Unknown Ligand (Human) B7-H5 including the enhancement or suppression of T cell proliferation, B7-H7 Unknown Receptor differentiation, and/or cytokine secretion.
    [Show full text]
  • Supplementary Table 1: Adhesion Genes Data Set
    Supplementary Table 1: Adhesion genes data set PROBE Entrez Gene ID Celera Gene ID Gene_Symbol Gene_Name 160832 1 hCG201364.3 A1BG alpha-1-B glycoprotein 223658 1 hCG201364.3 A1BG alpha-1-B glycoprotein 212988 102 hCG40040.3 ADAM10 ADAM metallopeptidase domain 10 133411 4185 hCG28232.2 ADAM11 ADAM metallopeptidase domain 11 110695 8038 hCG40937.4 ADAM12 ADAM metallopeptidase domain 12 (meltrin alpha) 195222 8038 hCG40937.4 ADAM12 ADAM metallopeptidase domain 12 (meltrin alpha) 165344 8751 hCG20021.3 ADAM15 ADAM metallopeptidase domain 15 (metargidin) 189065 6868 null ADAM17 ADAM metallopeptidase domain 17 (tumor necrosis factor, alpha, converting enzyme) 108119 8728 hCG15398.4 ADAM19 ADAM metallopeptidase domain 19 (meltrin beta) 117763 8748 hCG20675.3 ADAM20 ADAM metallopeptidase domain 20 126448 8747 hCG1785634.2 ADAM21 ADAM metallopeptidase domain 21 208981 8747 hCG1785634.2|hCG2042897 ADAM21 ADAM metallopeptidase domain 21 180903 53616 hCG17212.4 ADAM22 ADAM metallopeptidase domain 22 177272 8745 hCG1811623.1 ADAM23 ADAM metallopeptidase domain 23 102384 10863 hCG1818505.1 ADAM28 ADAM metallopeptidase domain 28 119968 11086 hCG1786734.2 ADAM29 ADAM metallopeptidase domain 29 205542 11085 hCG1997196.1 ADAM30 ADAM metallopeptidase domain 30 148417 80332 hCG39255.4 ADAM33 ADAM metallopeptidase domain 33 140492 8756 hCG1789002.2 ADAM7 ADAM metallopeptidase domain 7 122603 101 hCG1816947.1 ADAM8 ADAM metallopeptidase domain 8 183965 8754 hCG1996391 ADAM9 ADAM metallopeptidase domain 9 (meltrin gamma) 129974 27299 hCG15447.3 ADAMDEC1 ADAM-like,
    [Show full text]
  • CD Markers Are Routinely Used for the Immunophenotyping of Cells
    ptglab.com 1 CD MARKER ANTIBODIES www.ptglab.com Introduction The cluster of differentiation (abbreviated as CD) is a protocol used for the identification and investigation of cell surface molecules. So-called CD markers are routinely used for the immunophenotyping of cells. Despite this use, they are not limited to roles in the immune system and perform a variety of roles in cell differentiation, adhesion, migration, blood clotting, gamete fertilization, amino acid transport and apoptosis, among many others. As such, Proteintech’s mini catalog featuring its antibodies targeting CD markers is applicable to a wide range of research disciplines. PRODUCT FOCUS PECAM1 Platelet endothelial cell adhesion of blood vessels – making up a large portion molecule-1 (PECAM1), also known as cluster of its intracellular junctions. PECAM-1 is also CD Number of differentiation 31 (CD31), is a member of present on the surface of hematopoietic the immunoglobulin gene superfamily of cell cells and immune cells including platelets, CD31 adhesion molecules. It is highly expressed monocytes, neutrophils, natural killer cells, on the surface of the endothelium – the thin megakaryocytes and some types of T-cell. Catalog Number layer of endothelial cells lining the interior 11256-1-AP Type Rabbit Polyclonal Applications ELISA, FC, IF, IHC, IP, WB 16 Publications Immunohistochemical of paraffin-embedded Figure 1: Immunofluorescence staining human hepatocirrhosis using PECAM1, CD31 of PECAM1 (11256-1-AP), Alexa 488 goat antibody (11265-1-AP) at a dilution of 1:50 anti-rabbit (green), and smooth muscle KD/KO Validated (40x objective). alpha-actin (red), courtesy of Nicola Smart. PECAM1: Customer Testimonial Nicola Smart, a cardiovascular researcher “As you can see [the immunostaining] is and a group leader at the University of extremely clean and specific [and] displays Oxford, has said of the PECAM1 antibody strong intercellular junction expression, (11265-1-AP) that it “worked beautifully as expected for a cell adhesion molecule.” on every occasion I’ve tried it.” Proteintech thanks Dr.
    [Show full text]
  • Supplemental Figures 032819.Pptx
    Summary of Supplemental Figures and Tables 1) Supplemental Figure 1. Frequency of phenotypically defined endothelial cells is unchanged in aged mice. 2) Supplemental Figure 2. Aged LSKs have increased myeloid/megakaryocytic bias 3) Supplemental Figure 3. RBCs are decreased and Platelets are increased in peripheral blood of aged mice 4) Supplemental Figure 4. Aged BMME cultures have increased MSC populations. 5) Supplemental Figure 5. Aged BMME cells increase young LSK cell engraftment following competitive transplantation 6) Supplemental Figure 6. Flow cytometry gating strategy for sorting of young and aged murine Mφs 7) Supplemental Figure 7. Flow cytometry gating strategy for sorting of human Mφs 8) Supplemental Figure 8. Axl-/- LT-HSCs have increased cell engraftment following competitive transplantation 9) Supplemental Figure 9. Schematic representation of mechanisms by which age-dependent defects in marrow Mφs induce megakaryocytic bias in HSC. 10) Table S1. List of antibodies used in the flow cytometric and cell sorting analyses described 11) Table S2. List of cell populations analyzed and their respective immunophenotypes 12) Table S3. List of top GO-BP categories enriched for significantly upregulated genes from murine aged vs young marrow macrophages 13) Table S4. List of significant KEGG Pathways enriched for significantly upregulated genes from murine aged vs young marrow macrophages 1 A Sinusoidal Arteriolar BV786 – CD31 Sca1 – PE-Cy7 B C Arteriolar EC Sinusoidal ECs (Lin-, CD45-, CD31+ Sca1+) (CD45-Lin-CD31+Sca1-) 0.08 0.15 0.06 0.10 0.04 0.05 0.02 PercentLiveof Cells PercentLiveof Cells 0.00 0.00 Young Aged Young Aged Supplemental Figure 1.
    [Show full text]
  • 2B4 (CD244) Signaling by Recombinant Antigen-Specific Chimeric
    Published OnlineFirst July 28, 2009; DOI: 10.1158/1078-0432.CCR-08-2810 Cancer Therapy: Preclinical 2B4 (CD244) Signaling by Recombinant Antigen-specific Chimeric Receptors Costimulates Natural Killer Cell Activation to Leukemia and Neuroblastoma Cells Bianca Altvater,1 Silke Landmeier,1 Sibylle Pscherer,1 Jaane Temme,1 Katharina Schweer,1 Sareetha Kailayangiri,1 Dario Campana,3 Heribert Juergens,1 Martin Pule,2 and Claudia Rossig1 Abstract Purpose: Novel natural killer (NK) cell–directed strategies in cancer immunotherapy aim at specifically modulating the balance between NK cell receptor signals toward tu- mor-specific activation. The signaling lymphocyte activation molecule–related receptor 2B4 (CD244) is an important regulator of NK cell activation. We investigated whether 2B4-enhanced activation signals can redirect the cytolytic function of human NK cells to NK cell–resistant and autologous leukemia and tumor targets. Experimental Design: In vitro–stimulated NK cells from healthy donors and pediatric leukemia patients were gene modified with CD19 or GD2-specific chimeric receptors containing either the T-cell receptor ζ or 2B4 endodomain alone or combined. Results: Chimeric 2B4 signaling alone failed to induce interleukin-2 receptor up-regula- tion and cytokine secretion but triggered a specific degranulation response. Integration of the 2B4 endodomain into T-cell receptor ζ chimeric receptors significantly enhanced all aspects of the NK cell activation response to antigen-expressing leukemia or neuro- blastoma cells, including CD25 up-regulation, secretion of IFN-γ and tumor necrosis factor-α, release of cytolytic granules, and growth inhibition, and overcame NK cell re- sistance of autologous leukemia cells while maintaining antigen specificity.
    [Show full text]
  • Impaired Activating Receptor Expression Pattern in Natural Killer Cells from Patients with Multiple Myeloma
    Letters to the Editor 732 specificity for the CD20 molecule for potential use in adoptive by IMF SC 110212, University of Muenster to CS. MP and CS immunotherapy of B-cell malignancies. We show that cTCR þ contributed equally. CD8 þ NKT cells can be expanded in large quantities and are M Pieper1, C Scheffold1, S Duwe2, C Rossig2, G Bisping1, highly effective in lysing CD20 molecule expressing Daudi and 1 3 2 1 1 Raji tumor cell lines, as well as freshly isolated malignant B M Stelljes , TF Tedder , H Jurgens , WE Berdel and J Kienast 1Department of Medicine/Hematology and Oncology, lymphocytes from B-CLL patients. In competitive targeting University of Muenster, Muenster, Germany; studies, we were able to dissect the major pathways of tumor 2 þ Department of Pediatric Hematology and Oncology, cell recognition and found that endowing CD8 NKT cells with University of Muenster, Muenster, Germany and CD20z chimeric receptors resulted in a significant increase of 3Department of Immunology, Duke University, cytotoxic activity against CD20 þ Daudi targets as compared to Durham, NC, USA nontransduced effectors. E-mail: [email protected] We also demonstrate that chimeric receptor mediated cytotoxicity is dependent on the expression level of the target antigen. Tumor targets expressing high levels of CD20 molecule (Daudi) were lysed more efficiently than low level expressing References targets (Raji). Of importance, cytotoxicity of ex vivo expanded cTCR þ CD8 þ NKT cells was more potent as compared to 1 Eshhar Z, Waks T, Gross G, Schindler DG. Specific activation and ex vivo expanded cTCR þ CD8 þ T cells.
    [Show full text]
  • Cx3cr1 Mediates the Development of Monocyte-Derived Dendritic Cells During Hepatic Inflammation
    CX3CR1 MEDIATES THE DEVELOPMENT OF MONOCYTE-DERIVED DENDRITIC CELLS DURING HEPATIC INFLAMMATION. Supplementary material Supplementary Figure 1: Liver CD45+ myeloid cells were pre-gated for Ly6G negative cells for excluding granulocytes and HDCs subsequently analyzed among the cells that were CD11c+ and had high expression of MHCII. Supplementary Table 1 low/- high + Changes in gene expression between CX3CR1 and CX3CR1 CD11b myeloid hepatic dendritic cells (HDCs) from CCl4-treated mice high Genes up-regulated in CX3CR1 HDCs Gene Fold changes P value Full name App 4,01702 5,89E-05 amyloid beta (A4) precursor protein C1qa 9,75881 1,69E-22 complement component 1, q subcomponent, alpha polypeptide C1qb 9,19882 3,62E-20 complement component 1, q subcomponent, beta polypeptide Ccl12 2,51899 0,011769 chemokine (C-C motif) ligand 12 Ccl2 6,53486 6,37E-11 chemokine (C-C motif) ligand 2 Ccl3 4,99649 5,84E-07 chemokine (C-C motif) ligand 3 Ccl4 4,42552 9,62E-06 chemokine (C-C motif) ligand 4 Ccl6 3,9311 8,46E-05 chemokine (C-C motif) ligand 6 Ccl7 2,60184 0,009272 chemokine (C-C motif) ligand 7 Ccl9 4,17294 3,01E-05 chemokine (C-C motif) ligand 9 Ccr2 3,35195 0,000802 chemokine (C-C motif) receptor 2 Ccr5 3,23358 0,001222 chemokine (C-C motif) receptor 5 Cd14 6,13325 8,61E-10 CD14 antigen Cd36 2,94367 0,003243 CD36 antigen Cd44 4,89958 9,60E-07 CD44 antigen Cd81 6,49623 8,24E-11 CD81 antigen Cd9 3,06253 0,002195 CD9 antigen Cdkn1a 4,65279 3,27E-06 cyclin-dependent kinase inhibitor 1A (P21) Cebpb 6,6083 3,89E-11 CCAAT/enhancer binding protein (C/EBP),
    [Show full text]
  • Understanding the Synergy of Nkp46 and Co-Activating Signals in Various NK Cell Subpopulations: Paving the Way for More Successful NK-Cell-Based Immunotherapy
    cells Article Understanding the Synergy of NKp46 and Co-Activating Signals in Various NK Cell Subpopulations: Paving the Way for More Successful NK-Cell-Based Immunotherapy 1,2, 3, 1, 1 Loris Zamai * , Genny Del Zotto y , Flavia Buccella y, Sara Gabrielli , Barbara Canonico 1 , Marco Artico 4, Claudio Ortolani 1 and Stefano Papa 1 1 Department of Biomolecular Sciences, University of Urbino “Carlo Bo”, 61032 Urbino, Italy; fl[email protected] (F.B.); [email protected] (S.G.); [email protected] (B.C.); [email protected] (C.O.); [email protected] (S.P.) 2 INFN-Gran Sasso National Laboratory, Assergi, 67100 L’Aquila, Italy 3 Area Aggregazione Servizi e Laboratori Diagnostici, IRCCS Istituto Giannina Gaslini, 16147 Genoa, Italy; [email protected] 4 Department of Sensory Organs, Sapienza University of Rome, 00161 Rome, Italy; [email protected] * Correspondence: [email protected]; Tel.: +39-0722-304319; Fax: +39-0722-304319 These authors contributed equally. y Received: 11 February 2020; Accepted: 13 March 2020; Published: 19 March 2020 Abstract: The NK cell population is characterized by distinct NK cell subsets that respond differently to the various activating stimuli. For this reason, the determination of the optimal cytotoxic activation of the different NK cell subsets can be a crucial aspect to be exploited to counter cancer cells in oncologic patients. To evaluate how the triggering of different combination of activating receptors can affect the cytotoxic responses of different NK cell subsets, we developed a microbead-based degranulation assay. By using this new assay, we were able to detect CD107a+ degranulating NK cells even within the less cytotoxic subsets (i.e., resting CD56bright and unlicensed CD56dim NK cells), thus demonstrating its high sensitivity.
    [Show full text]
  • 2B4 (CD244)–CD48 Interactions Provide a Novel MHC Class I–Independent System for NK-Cell Self-Tolerance in Mice
    From www.bloodjournal.org by guest on July 30, 2018. For personal use only. IMMUNOBIOLOGY Brief report 2B4 (CD244)–CD48 interactions provide a novel MHC class I–independent system for NK-cell self-tolerance in mice Megan E. McNerney, Dustin Guzior, and Vinay Kumar Natural killer (NK) cells must be able to that 2B4 (CD244) and CD48 represent a the first in vivo evidence for MHC-indepen- eliminate infected and transformed cells second system for murine NK-cell self- dent NK self-recognition in a bone mar- while remaining tolerant of normal cells. recognition. 2B4 and MHC class I recep- row rejection assay. These data suggest NK-cell self-tolerance is thought to be tors act nonredundantly to inhibit NK that NK-cell self-tolerance can be medi- maintained by self–major histocompatibil- lysis of syngeneic tumor cells. NK cells ated by molecules other than MHC. ity complex (MHC) class I recognition; from ␤2 microglobulin (␤2m)–deficient (Blood. 2005;106:1337-1340) however, there are examples where NK mice and NK cells that lack expression of cells are not regulated by MHC class I and self-MHC–binding inhibitory receptors are yet remain self-tolerant. Here, we show inhibited by 2B4. Moreover, we provide © 2005 by The American Society of Hematology Introduction B and T lymphocytes from healthy individuals are tolerant of MHC class I expression due to transporter-associated with antigen self-antigens and are activated by foreign antigens. In contrast, processing 2 (TAP-2) protein or ␤2 microglobulin (␤2m) defi- although natural killer (NK) cells are also tolerant of normal cells ciency, have self-tolerant NK cells.10 NK cells from MHC class that express self-ligands, NK cells are activated by cells that lack I–deficient individuals may have decreased activating receptor self-ligands.
    [Show full text]
  • NK Cells: Receptors and Functions Eric Vivier and Sophie Ugolini
    NK cells: receptors and functions Eric Vivier and Sophie Ugolini Natural killer (NK) cells were identified in 1975 as lymphocytes of the innate many processes that were originally restricted to adaptive immunity, such immune system that can kill tumour cells. Since then, NK cells have been as priming, education and memory, are now known to occur in NK cells. shown to kill an array of ‘stressed’ cells and secrete cytokines that Indeed, NK cells undergo sophisticated processes of adaptation that allow participate in shaping adaptive immune responses. A key feature of NK cells them to be tuned to their environment. There is also a growing interest in resides in their capacity to distinguish stressed cells (such as tumour cells, manipulating NK cells in innovative therapeutic settings. For example, the infected cells and damaged cells) from normal cells. Although NK cells are understanding of NK cell inhibition by MHC class I-specific receptors has generally considered to be components of early innate immune defence, prompted the design of innovative anticancer therapies. The NK cell detection system includes numerous receptors, Key receptors on human NK cells the engagement of which dictates the quality and intensity Inhibitory receptors Activating receptors, adhesion molecules or co-stimulatory molecules of the NK cell response. NK cells use inhibitory receptors to gauge the absence of constitutively expressed self molecules Target cell Nectin 2 SLAMF7 Sialic CLEC2D (CD112, PVRL2) NECL2 L-selectin NECL5 AICL NTB-A (CRACC, on susceptible target cells. As a consequence, NK cells can Receptors for MHC class I and class I-Iike molecules B7-H6 acid Cadherins (LLT1) CMV pp65 NECL5 (CADM1) (CD62L) (CD155, PVR) (CLEC2B) CEACAM1 (SLAMF6) CS1, CD319) recognize ‘missing self’ on haematopoietic cells.
    [Show full text]