DOCSLIB.ORG

MGD011, a CD19 X CD3 Dual-Affinity Retargeting Bi-Specific Molecule Incorporating Extended Circulating Half-Life for the Treatment of B-Cell Malignancies

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
MGD011, a CD19 X CD3 Dual-Affinity Retargeting Bi-Specific Molecule Incorporating Extended Circulating Half-Life for the Treatment of B-Cell Malignancies Published OnlineFirst September 23, 2016; DOI: 10.1158/1078-0432.CCR-16-0666 Cancer Therapy: Preclinical Clinical Cancer Research MGD011, A CD19 x CD3 Dual-Affinity Retargeting Bi-specific Molecule Incorporating Extended Circulating Half-life for the Treatment of B-Cell Malignancies Liqin Liu, Chia-Ying K. Lam, Vatana Long, Lusiana Widjaja, Yinhua Yang, Hua Li, Linda Jin, Steve Burke, Sergey Gorlatov, Jennifer Brown, Ralph Alderson, Margaret D. Lewis, Jeffrey L. Nordstrom, Scott Koenig, Paul A. Moore, Syd Johnson, and Ezio Bonvini Abstract Purpose: CD19, a B-cell lineage-specific marker, is highly autologous B-cell depletion in PBMCs from both species. represented in B-cell malignancies and an attractive target for MGD011-mediated killing was accompanied by target-depen- therapeutic interventions. MGD011 is a CD19 x CD3 DART dent T-cell activation and expansion, cytokine release and bispecific protein designed to redirect T lymphocytes to eliminate upregulation of perforin and granzyme B. MGD011 demon- CD19-expressing cells. MGD011 has been engineered with a strated antitumor activity against localized and disseminated modified human Fc domain for improved pharmacokinetic (PK) lymphoma xenografts reconstituted with human PBMCs. In properties and designed to cross-react with the corresponding cynomolgus monkeys, MGD011 displayed a terminal half-life antigens in cynomolgus monkeys. Here, we report on the preclin- of 6.7 days; once weekly intravenous infusion of MGD011 at ical activity, safety and PK properties of MGD011. dosesupto100mg/kg, the highest dose tested, was well Experimental Design: The activity of MGD011 was evaluated tolerated and resulted in dose-dependent, durable decreases in several in vitro and in vivo models. PK, safety and pharmaco- in circulating B cells accompanied by profound reductions of B dynamic activity was also assessed in dose-escalation and repeat- lymphocytes in lymphoid organs. dose studies of MGD011 administered once weekly in cynomol- Conclusions: The preclinical activity, safety and PK pro- gus monkeys. file support clinical investigation of MGD011 as a thera- Results: MGD011 mediated killing of human B-cell lympho- peutic candidate for the treatment of B-cell malignancies. ma lines by human or cynomolgus monkey PBMCs as well as Clin Cancer Res; 1–13. Ó2016 AACR. Introduction improved outcome in advanced B-cell malignancies. Yet, approximately 20,000 patients die of lymphoma every year in B-cell malignancies represent a heterogeneous group of dis- the United States alone. orders with varying characteristics and clinical behaviors (1). Providing T lymphocytes (CTL) with the ability to recognize Although systemic chemotherapy is still the mainstay of treat- and destroy tumor cells has shown promise in advanced forms ment for B-cell malignancies, kinase inhibitors that selectively of leukemia and lymphoma. In the form of chimeric antigen target molecules at the core of the transformation process and receptor (CAR) T-cell therapy, such an approach requires ex vivo antibody therapy are now well established tools (2). Among the isolation, transduction, and reinfusion of the patient's T cells. latter category, rituximab (Rituxan), a monoclonal antibody This complexity can be overcome with bispecific antibodies (mAb) that targets the B-cell antigen CD20, induces direct that bind simultaneously to an antigen expressed by malignant tumor cell apoptosis as well as complement- and antibody- BcellsandanactivationmoleculeonTlymphocytes(2).The dependent cytotoxicity (2, 3). These orthogonal mechanisms of pan B-cell marker, CD19, has emerged as a promising antigen action form the basis for therapeutic combinations that have for targeting B-cell malignancies because of its broader expres- sion profile and lower rate of downregulation compared with other B-cell antigens (3). Its expression is highly conserved in Research, MacroGenics, Inc., Rockville, Maryland. the majority of B-cell tumors (4), with normal to high levels of Note: Supplementary data for this article are available at Clinical Cancer expression in 80% of acute lymphoblastic leukemia (ALL), 88% Research Online (http://clincancerres.aacrjournals.org/). of B-cell lymphomas, and all chronic lymphocytic leukemias Corresponding Author: Ezio Bonvini, MacroGenics Inc., 9704 Medical Center (CLL; refs. 5, 6). þ Drive, Rockville, MD 20850. Phone: 301-354-2638; Fax: 301-251-5321; E-mail: Redirection of CTL to CD19 leukemia cells via the bispecific [email protected] T-cell engager (BiTE) blinatumomab (Blincyto; ref. 7) is effec- doi: 10.1158/1078-0432.CCR-16-0666 tive in patients with B-cell malignancies whose disease did not Ó2016 American Association for Cancer Research. respondtostandardchemo-immunotherapies and has been www.aacrjournals.org OF1 Downloaded from clincancerres.aacrjournals.org on October 1, 2021. © 2016 American Association for Cancer Research. Published OnlineFirst September 23, 2016; DOI: 10.1158/1078-0432.CCR-16-0666 Liu et al. FcgR and C1q binding (14). Control molecules in which the Translational Relevance variable domain sequences of an anti-fluorescein mAb 4-4-20 Progress has been made in the clinical management of B-cell (15) replaced either of the DART protein arms (Fluo x CD3 or neoplasms, although most remain ultimately incurable with CD19 x Fluo) were engineered in a similar manner. The DART current modalities. Redirecting T lymphocytes to lyse lympho- proteins were expressed transiently in CHO-S cells (8) and puri- ma/leukemia cells via bispecific molecules that simultaneous- fied to greater than 99% purity using protein A and either size- ly engage CD3 on T cells with a B-cell antigen, such as CD19, exclusion chromatography (SEC) or other polishing steps. The has emerged as a powerful novel concept, highlighted by the purified DARTs have very low levels (less than 1%) of high clinical success of blinatumomab (Blincyto). Blinatumomab, molecular weight (HMW) protein present and have an apparent however, requires continuous infusion, owing to its short molecular weight of approximately 110 kDa (Supplementary Fig. circulating half-life. We report here on the preclinical devel- S1A and B). opment of MGD011, a bispecific DART molecule with increased in vitro cytolytic activity compared to blinatumomab Other reagents, cell lines, and tissue samples and engineered for improved circulating half-life. MGD011 Recombinant soluble human and cynomolgus CD3e/d chi- showed potent antitumor activity in mouse leukemia/lym- meric proteins, as well as human and cynomolgus CD19 and phoma models and displayed prolonged pharmacokinetic CD19-His proteins, were expressed in CHO-S cells. MOLM-13 properties in cynomolgus monkeys, a cross-reacting species. and JIMT-1 were obtained from DSMZ (Braunschweig, Ger- MGD011 was well tolerated in monkeys, with durable and many); Jeko-1 cells were from the ATCC and HBL-2 from the profound B-cell depletion following weekly administrations. NationalInstitutesofHealth(Bethesda,MD;ref.16).Raji/GF,a þ MGD011's potent activity and pharmacokinetic properties CD19 Burkitt's B-cell lymphoma expressing luciferase and may offer therapeutic convenience and applicability in the green fluorescent protein by stable transfection, was established treatment of B-cell malignancies. at MacroGenics. All cell lines were passaged for less than 3 months after thawing; all lines were confirmed to be free of mycoplasma by PCR (Taconic, 2013) and were authenticated on the basis of morphology, growth characteristics and CD19 approved by the FDA for the treatment of patients with Phi- expression. Heparinized human whole blood was from Bio- fi ladelphia chromosome-negative relapsed or refractory B-cell logical Specialty Corporation. Cryopreserved puri ed primary precursor ALL. DART proteins are bispecific, antibody-based CLL patient samples were from AllCells, LLC. Heparinized molecules with favorable stability, manufacturability, and whole blood from cynomolgus monkeys was from Worldwide potency; furthermore, a CD19 x CD3 DART protein compared Primates, Inc. favorably with a BiTE of the same pair of VH and VL sequences in redirected cytolysis assays (8, 9). MGD011 (also known as Binding studies JNJ-64052781) is another CD19 x CD3 DART protein designed MGD011 binding to human or cynomolgus monkey CD3 or þ to simultaneously target CD19 cells for recognition and elim- CD19 proteins was analyzed by ELISA or surface plasmon reso- nance (SPR) as previously described (13); binding to primary ination by CD3-expressing T lymphocytes as effector cells. þ þ þ MGD011 was engineered with a human immunoglobulin human or cynomolgus monkey CD20 , CD4 or CD8 cells was fl G1 (IgG1) Fc domain to bind the neonatal Fc receptor (FcRn) analyzed by ow cytometry. and engage the IgG salvage pathway, thus conferring prolonged circulating half-life and the resultant dosing convenience. To Cell killing assay avoid unintended (target independent) CD3-mediated T-cell For CTL assays, DART protein mediated killing of target cell fi activation via interaction with Fc gamma receptors (FcgR), the lines in the presence of human or cynomolgus PBMCs or puri ed Fc domain was mutated to greatly reduce or eliminate binding T cells was determined by lactate dehydrogenase (LDH) release or to these receptors as well as complement. Unlike blinatumo- luminescence assays as previously described (17). mab, which reacts only with human or chimpanzee's antigens (10), MGD011 cross-reacts with both CD19 and CD3 mole- B-cell depletion assay cules in macaques, enabling preclinical evaluation in a relevant
Load more

Recommended publications
  • 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]
  • T Cells the Usual Subsets
    T cells: the usual subsets Chen Dong and Gustavo J. Martinez T cells have important roles in immune responses and function by directly secreting soluble mediators or important for adaptation of immune responses in different microenvironments and might be particularly through cell contact-dependent mechanisms. Many T cell subsets have been characterized. Although relevant for host defence against pathogens that colonize different tissues. Distinct T cell subsets, or effector T cells were originally considered to be terminally differentiated, a growing body of evidence has differentiation states, can be identified based on the cell surface markers expressed and/or the effector challenged this view and suggested that the phenotype of effector T cells is not completely fixed but is molecules produced by a particular T cell population. This Poster summarizes our current understanding of more flexible or plastic. T cells can have ‘mixed’ phenotypes (that is, have characteristics usually the surface markers, transcriptional regulators, effector molecules and functions of the different T cell associated with more than one T cell subset) and can interconvert from one subset phenotype to another, subsets that participate in immune responses. Further knowledge of how these T cell subsets are regulated IMMUNOLOGY although instructive signalling can lead to long-term fixation of cytokine memory. T cell plasticity can be and cooperate with each other will provide us with better tools to treat immune-related diseases. Cytotoxic T cell Exhausted T cell
    [Show full text]
  • Optimal Minimal Panels of Immunohistochemistry for Diagnosis of B-Cell Lymphoma for Application in Countries with Limited Resources and for Triaging Cases Before
    AJCP /ORIGINAL ARTICLE Optimal Minimal Panels of Immunohistochemistry for Diagnosis of B-Cell Lymphoma for Application in Countries With Limited Resources and for Triaging Cases Before Referral to Specialist Centers Downloaded from https://academic.oup.com/ajcp/article-abstract/145/5/687/2195691 by World Health Organization user on 09 January 2019 Maria Giulia Disanto, MD,1 Maria Raffaella Ambrosio, MD, PhD,2 Bruno Jim Rocca, MD, PhD,2 Hazem A. H. Ibrahim, FRCPath, PhD,1,3 Lorenzo Leoncini, MD, PhD,2 and Kikkeri N. Naresh, MD, FRCPath1 From the 1Department of Histopathology, Imperial College Healthcare NHS Trust & Imperial College, London, United Kingdom; 2Department of Medical Biotechnologies, Section of Pathology, University of Siena, Siena, Italy; and 3Department of Histopathology, Faculty of Medicine, Mansoura University, Mansoura, Egypt. Key Words: Lymphoma; B-cell lymphoma; Immunohistochemistry; Diagnosis; Classification; Developing countries Am J Clin Pathol May 2016;145:687-695 DOI: 10.1093/AJCP/AQW060 ABSTRACT Lymphomas are a collection of different malignancies “arising” from lymphoid cells. They include about 49 entities, Objectives: Establish and validate optimal minimal and over 19 provisional entities and subsets.1 About 85% of immunohistochemistry panels for usage in a staged lymphomas are of B-cell origin. Precision in lymphoma diag- algorithmic manner for precise diagnosis of B-cell nosis requires expertise and infrastructure. The entities are lymphomas in countries with limited resources. Suggest defined based on morphology, immunohistochemistry (on short panels of immunostains to be used in referring units some occasions in situ hybridization), cytogenetics/fluores- that refer suspected lymphomas to specialist diagnostic cent in situ hybridization (FISH), molecular genetics and clin- centers in resourceful countries.
    [Show full text]
  • CD3+CD5+CD4-CD8-) Alpha/Beta T Cell Receptor-Bearing Cells
    Mlsa generated suppressor cells. I. Suppression is mediated by double-negative (CD3+CD5+CD4-CD8-) alpha/beta T cell receptor-bearing cells. This information is current as of October 2, 2021. M Bruley-Rosset, I Miconnet, C Canon and O Halle-Pannenko J Immunol 1990; 145:4046-4052; ; http://www.jimmunol.org/content/145/12/4046 Downloaded from Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision http://www.jimmunol.org/ • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription by guest on October 2, 2021 Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 1990 by American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. 0022-1767/90/14512-4046$02.00/0 THEJOURNAL OF IMMUNOLCGY Vol. 145.4046-4052. No. 12. December 15. 1990 Copyright 0 1990 by The American Association of lmmunologists Printed In U.S.A. Mls" GENERATED SUPPRESSORCELLS I. Suppression is Mediated by Double-Negative (CD3+CDVCD4-CD8-) a/@ T Cell Receptor-Bearing Cells' Grafting of cells from B10.D2 (H-2d)donors into H- The GVHR3 remains a major problem after bone mar- 2 compatible lethally irradiated (DBA/2 x B1O.DZ)Fl row transplantation, even in MHC-compatible donor-re- hostsresults in a severe graft-vs-host reaction cipient combinations.
    [Show full text]
  • Ligand for Bovine CD5 the Identification and Characterization of A
    The Identification and Characterization of a Ligand for Bovine CD5 Karen M. Haas and D. Mark Estes This information is current as J Immunol 2001; 166:3158-3166; ; of September 27, 2021. doi: 10.4049/jimmunol.166.5.3158 http://www.jimmunol.org/content/166/5/3158 Downloaded from References This article cites 46 articles, 18 of which you can access for free at: http://www.jimmunol.org/content/166/5/3158.full#ref-list-1 Why The JI? Submit online. http://www.jimmunol.org/ • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average by guest on September 27, 2021 Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2001 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Identification and Characterization of a Ligand for Bovine CD51 Karen M. Haas* and D. Mark Estes2*† CD5, a type I glycoprotein expressed by T cells and a subset of B cells, is thought to play a significant role in modulating Ag receptor signaling. Previously, our laboratory has shown that bovine B cells are induced to express this key regulatory molecule upon Ag receptor cross-linking.
    [Show full text]
  • Significant Enlargement of a Specific Subset of CD3+CD8+ Peripheral Blood Leukocytes Mediating Cytotoxic T-Lymphocyte Activity D
    Proc. Natl. Acad. Sci. USA Vol. 90, pp. 9427-9430, October 1993 Immunology Significant enlargement of a specific subset of CD3+CD8+ peripheral blood leukocytes mediating cytotoxic T-lymphocyte activity during human immunodeficiency virus infection (T lymphocyte subset/monoclonal antibody/AIDS) A. BENSUSSAN*t, C. RABIANt, V. SCHIAVON*, D. BENGOUFAt, G. LECA*, AND L. BOUMSELL* Institut Nationale de la Sant6 et de la Recherche Medicale U93, Association Claude Bernard, and tLaboratoire d'Immunologie et d'Histocompatibilite, H6pital Saint-Louis, 1 Avenue Claude Veliefaux, 75475 Paris cedex 10, France Communicated by Jean Dausset, July 20, 1993 ABSTRACT We have obtained a monoclonal antibody studies have shown that human immunodeficiency virus termed BY55 that defines an 80-kDa cell-surface structure on (HIV) infection significantly changes the phenotype of cir- a subset ofcirculating peripheral blood mononucleocytes. This culating lymphocytes and increases CD8 T lymphocytes (19), structure, which was not detected on most cell lines or activated it was interesting to look for this cytotoxic CD3+CD8+BY55+ lymphocytes, was expressed exclusively on 15-25% of CD2+ circulating cell subset in asymptomatic HIV-positive individ- circulating lymphocytes, including a major subset within the uals with various numbers of CD4 cells. Results indicated CD3- and the T-cell receptor y6+ lymphocytes and a small that peripheral blood CD3+CD8+BY55+ cells significantly percentage of the CD3+CD8+ cells. Moreover, we have shown increased in HIV-positive individuals. that the BY55 molecule delineated the competent killer circu- lating lymphocytes. In the present report, additional two- and MATERIALS AND METHODS three-color immunofluorescence studies of peripheral blood lymphocytes were done to precisely determine BY55 expression mAbs.
    [Show full text]
  • Immunological Synapse Formation Licenses CD40-CD40L Accumulations at T-APC Contact Sites1
    The Journal of Immunology Immunological Synapse Formation Licenses CD40-CD40L Accumulations at T-APC Contact Sites1 Judie Boisvert, Samuel Edmondson, and Matthew F. Krummel2 The maintenance of tolerance is likely to rely on the ability of a T cell to polarize surface molecules providing “help” to only specific APCs. The formation of a mature immunological synapse leads to concentration of the TCR at the APC interface. In this study, we show that the CD40-CD154 receptor-ligand pair is also highly concentrated into a central region of the synapse on mouse lymphocytes only after the formation of the TCR/CD3 c-SMAC. Concentration of this ligand was strictly dependent on TCR recognition, the binding of ICAM-1 to T cell integrins and the presence of an intact cytoskeleton in the T cells. This may provide a novel explanation for the specificity of T cell help directing the help signal to the site of Ag receptor signal. It may also serve as a site for these molecular aggregates to coassociate and/or internalize alongside other signaling receptors. The Journal of Im- munology, 2004, 173: 3647–3652. uring the onset and propagation of immune responses, T Interactions of T cells with Ag-bearing APCs is associated with cells send and receive signals via Ag-dependent and Ag- a coalescence of TCRs, other related receptors such as CD2, CD4, independent ligands. Whereas peptide-class II MHC CD28, and signaling molecules such as p56lck, fyn, linker for ac- D ϩ complexes on APC are recognized by the TCR on CD4 helper T tivation of T cells, and protein kinase C␪ into the central portion of cells, separate cell surface receptors modulate T cell-APC inter- an “immunological synapse” (15–23).
    [Show full text]
  • A 39-Kda Protein on Activated Helper T Cells Binds CD40 and Transduces the Signal for Cognate Activation of B Cells RANDOLPH J
    Proc. Natl. Acad. Sci. USA Vol. 89, pp. 6550-6554, July 1992 Immunology A 39-kDa protein on activated helper T cells binds CD40 and transduces the signal for cognate activation of B cells RANDOLPH J. NOELLE*t, MEENAKSHI RoY*, DAVID M. SHEPHERD*, IVAN STAMENKOVICO, JEFFREY A. LEDBETTER§, AND ALEJANDRO ARUFFO§ *Department of Microbiology, Dartmouth Medical School, One Medical Center Drive, Lebanon, NH 03756; *Bristol-Myers Squibb Pharmaceutical Research Institute, Seattle, WA 98121; and *Department of Pathology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114 Communicated by Leon E. Rosenberg, April 22, 1992 (receivedfor review February 5, 1992) ABSTRACT CD40 is a B-cell surface molecule that has MATERIALS AND METHODS been shown to induce B-cell growth upon ligation with mono- clonal antibodies. This report shows that triggering via CD40 Mice. Female DBA/2J mice (The Jackson Laboratory) is essential for the activation of resting B cells by helper T cells were used for the preparation of filler cells to support the (Th). A soluble fusion protein of CD40 and human immuno- growth of Th clones and in the preparation of resting B cells. globulin, CD40-Ig, inhibited the induction ofB-cell cycle entry, Th Clones. D1.6, an I-Ad-restricted, rabbit Ig-specific ThO proliferation, and differentiation by activated Thi and Tb2. clone, and CDC35, an I-Ad-restricted, rabbit Ig-specific Th2 The ligand for CD40 was identified as a 39-kDa membrane clone, were obtained from David Parker (University of protein that was selectively expressed on activated Tb. A Massachusetts, Worcester). In this paper, D1.6 will be re- monoclonal antibody specific for the 39-kDa protein inhibited ferred to as Thl and CDC35 as Tb2 (7).
    [Show full text]
  • Features of Human CD3+CD20+ T Cells
    Published July 13, 2016, doi:10.4049/jimmunol.1600089 The Journal of Immunology Features of Human CD3+CD20+ T Cells Elisabeth Schuh,*,† Kerstin Berer,‡ Matthias Mulazzani,x,{ Katharina Feil,x Ingrid Meinl,* Harald Lahm,‖ Markus Krane,‖,# Rudiger€ Lange,‖,# Kristina Pfannes,** Marion Subklewe,** Robert Gurkov,€ †† Monika Bradl,† Reinhard Hohlfeld,*,‡‡ Tania Kumpfel,*€ Edgar Meinl,*,1 and Markus Krumbholz*,1,2 Monoclonal Abs against CD20 reduce the number of relapses in multiple sclerosis (MS); commonly this effect is solely attributed to depletion of B cells. Recently, however, a subset of CD3+CD20+ T cells has been described that is also targeted by the anti-CD20 mAb rituximab. Because the existence of cells coexpressing CD3 and CD20 is controversial and features of this subpopulation are poorly understood, we studied this issue in detail. In this study, we confirm that 3–5% of circulating human T cells display CD20 on their surface and transcribe both CD3 and CD20. We report that these CD3+CD20+ T cells pervade thymus, bone marrow, and secondary lymphatic organs. They are found in the cerebrospinal fluid even in the absence of inflammation; in the cerebrospinal fluid of MS patients they occur at a frequency similar to B cells. Phenotypically, these T cells are enriched in CD8+ and CD45RO+ memory cells and in CCR72 cells. Functionally, they show a higher frequency of IL-4–, IL-17–, IFN-g–, and TNF-a–producing cells compared with T cells lacking CD20. CD20-expressing T cells respond variably to immunomodulatory treatments given to MS patients: they are reduced by fingolimod, alemtuzumab, and dimethyl fumarate, whereas natalizumab disproportionally increases them in the blood.
    [Show full text]
  • Laboratory Guidelines for Enumerating CD4 T Lymphocytes in the Context of HIV/AIDS SEA-HLM-392 Distribution: Limited
    Laboratory Guidelines for enumerating CD4 T Lymphocytes in the context of HIV/AIDS SEA-HLM-392 Distribution: Limited Laboratory Guidelines for enumerating CD4 T Lymphocytes in the context of HIV/AIDS Regional Office for South-East Asia New Delhi, June 2007 WHO Library Cataloguing-in-Publication Data World Health Organization, Regional Office for South-East Asia. Laboratory guidelines for enumerating CD4 T lymphocytes in the context of HIV/AIDS 1. T lymphocytes – methods 2. CD4-positive T lymphocytes – immunology 3. HIV infections – diagnosis 4. Quality assurance, health care 5. Laboratory techniques and procedures – methods 6. Flow cytometry – methods 7. Guidelines. ISBN 978-92-9022-298-9 (NLM classification: WH 200) © World Health Organization 2007 This document is not issued to the general public, and all rights are reserved by the World Health Organization (WHO). The document may not be reviewed, abstracted, quoted, reproduced or translated, in part or in whole, without the prior written permission of WHO. No part of this document may be stored in a retrieval system or transmitted in any form or by any means – electronic, mechanical or other – without the prior written permission of WHO. The mention of specific companies or of certain manufacturers’ products does not imply that they are endorsed or recommended by the World Health Organization in preference to others of a similar nature that are not mentioned. Errors and omissions excepted, the names of proprietary products are distinguished by initial capital letters. The views expressed in documents by named authors are solely the responsibility of those authors. Contents Page 1. Natural history of HIV infection ..........................................................................
    [Show full text]
  • Overcoming Challenges for CD3-Bispecific Antibody Therapy In
    cancers Review Overcoming Challenges for CD3-Bispecific Antibody Therapy in Solid Tumors Jim Middelburg 1 , Kristel Kemper 2, Patrick Engelberts 2 , Aran F. Labrijn 2 , Janine Schuurman 2 and Thorbald van Hall 1,* 1 Department of Medical Oncology, Oncode Institute, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands; [email protected] 2 Genmab, 3584 CT Utrecht, The Netherlands; [email protected] (K.K.); [email protected] (P.E.); [email protected] (A.F.L.); [email protected] (J.S.) * Correspondence: [email protected]; Tel.: +31-71-5266945 Simple Summary: CD3-bispecific antibody therapy is a form of immunotherapy that enables soldier cells of the immune system to recognize and kill tumor cells. This type of therapy is currently successfully used in the clinic to treat tumors in the blood and is under investigation for tumors in our organs. The treatment of these solid tumors faces more pronounced hurdles, which affect the safety and efficacy of CD3-bispecific antibody therapy. In this review, we provide a brief status update of this field and identify intrinsic hurdles for solid cancers. Furthermore, we describe potential solutions and combinatorial approaches to overcome these challenges in order to generate safer and more effective therapies. Abstract: Immunotherapy of cancer with CD3-bispecific antibodies is an approved therapeutic option for some hematological malignancies and is under clinical investigation for solid cancers. However, the treatment of solid tumors faces more pronounced hurdles, such as increased on-target off-tumor toxicities, sparse T-cell infiltration and impaired T-cell quality due to the presence of an Citation: Middelburg, J.; Kemper, K.; immunosuppressive tumor microenvironment, which affect the safety and limit efficacy of CD3- Engelberts, P.; Labrijn, A.F.; bispecific antibody therapy.
    [Show full text]
  • Lymphokine-Activated Killer Activity in Long-Term Cultures with Anti-CD3 Plus Interleukin 2: Identification and Isolation of Effector Subsets1
    (CANCER RESEARCH 49. 963-968. February 15. 1989] Lymphokine-activated Killer Activity in Long-Term Cultures with Anti-CD3 plus Interleukin 2: Identification and Isolation of Effector Subsets1 Augusto C. Ochoa,2 Diane E. Hasz, Rebecca Rezonzew, Peter M. Anderson, and Fritz H. Bach Immunobiology Research Center, University of Minnesota Hospital and Clinic, Minneapolis, Minnesota 55455 ABSTRACT short-term LAK cultures, while CD3+ cells from such cultures have low lytic activity against NK-resistant targets (8-11). Peripheral blood lymphocytes cultured in recombinant interleukin 2 We have recently reported the generation of large numbers during 3 to 5 days (short-term cultures) develop the ability to lyse natural of cells with LAK activity using long-term (14 to 21 day) killer-resistant tumor lines and fresh tumor cells, i.e., express lympho- cultures of cells stimulated with anti-CD3 (OKT3) moAb and kine-activated killer (LAR) function. Phenotypic analysis has shown these cells to be natural killer cells, i.e., CD16+ and/or Leu 19+ cells. IL2 (6). Cells stimulated with OKT3 + IL2 undergo a high CD3*,CD16~ T-cells, instead, develop very low LAK function in these increase in cell number while maintaining specific LAK activity cultures. comparable to that of cells cultured for 3 to 5 days. LAK We recently reported the development of long-term (up to 21 days) activity of cells stimulated with OKT3 + IL2 for 14 days is cultured cells with LAK activity by stimulation with OKT3 + interleukin significantly increased when incubated for the last 48 h of 2 (11.2).These culture conditions repeatedly resulted in a several hundred culture in medium containing ILl-ß,IFN-7, or -ßinaddition fold expansion in cell number.
    [Show full text]