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Effector Memory αβ T Lymphocytes Can Express Fc γRIIIa and Mediate Antibody-Dependent Cellular Cytotoxicity This information is current as Béatrice Clémenceau, Régine Vivien, Mathilde Berthomé, of September 27, 2021. Nelly Robillard, Richard Garand, Géraldine Gallot, Solène Vollant and Henri Vié J Immunol 2008; 180:5327-5334; ; doi: 10.4049/jimmunol.180.8.5327 http://www.jimmunol.org/content/180/8/5327 Downloaded from References This article cites 43 articles, 21 of which you can access for free at: http://www.jimmunol.org/content/180/8/5327.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • 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 by guest on September 27, 2021 *average 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 © 2008 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Effector Memory ␣ T Lymphocytes Can Express Fc␥RIIIa and Mediate Antibody-Dependent Cellular Cytotoxicity1 Be´atrice Cle´menceau,*† Re´gine Vivien,*† Mathilde Berthome´,*† Nelly Robillard,‡ Richard Garand,‡ Ge´raldine Gallot,*† Sole`ne Vollant,*† and Henri Vie´2*† Human memory T cells are comprised of distinct populations with different homing potential and effector functions: central memory T cells that mount recall responses to Ags in secondary lymphoid organs, and effector memory T cells that confer immediate protection in peripheral tissues. In the present study we demonstrate that a proportion of effector memory T cells express Fc␥RIIIa (CD16), are perforin positive, and directly mediate Ab-dependent cytotoxicity ex vivo. This particular ␣ T lymphocyte subset has the morphology of large granular lymphocytes, increases proportionately in vivo during reactive lympho- cytosis, and can be detected in vitro among EBV-specific T lymphocytes after stimulation with EBV Ags. Consequently, during a normal immune response, amplification of these effector memory T lymphocytes that are capable of Ab-dependent cytotoxicity Downloaded from may have beneficial or harmful consequences depending on the presence of pathogen- or tissue-specific Abs, respectively. The Journal of Immunology, 2008, 180: 5327–5334. he vast majority of cells that express the IgG FcR re- tionated T cell population with respect to reactivity with anti-CD3, sponsible for Ab-dependent cellular cytotoxicity -CD4, and -CD5 mAb (OKT3, OKT4, and OKT5, respectively) 3 T (ADCC ;Fc␥RIIIa or CD16) belong to the innate im- while the T␥-enriched population contained, in fact, only 10–20% http://www.jimmunol.org/ mune system (1). These include monocytes/macrophages and of T cells as defined by OKT3 (10–12). The authors of these stud- NK cells. Among PBMC, it is assumed that essentially all ies concluded appropriately that the T␥ cells included a small pop- ADCC activity is mediated by NK cells. Nevertheless, the ex- ulation reactive with OKT3 and a large population reactive with istence of T lymphocytes bearing FcRs for Ig has long been OKM1, most likely corresponding to NK cells (10). Based on recognized, although data concerning ADCC mediated by T these findings, it was unclear whether ADCC attributed to the T␥ lymphocytes are rare. subset was mediated by the CD3- (non-T) NK fraction or the small Indeed, in the past, initial attempts to discriminate subsets proportion of CD3ϩ T cells that express FcR for IgG. Later on, as among T lymphocytes relied on the use of either antisera or dif- stated above, essentially all NK activity present among PBMC was by guest on September 27, 2021 ferential FcR binding. Using the latter approach, two human T cell attributed to the NK population. In the early 1980s, several mAb subsets could be separated: those capable of binding the Fc portion were produced (anti-Leu-11, VEP 13, B73.1, and 3G8) that spe- of IgM (T ) and those capable of binding the Fc portion of IgG cifically reacted with the IgG FcR responsible for ADCC ␥ (T ) (2–6). However, at that time, discrimination between differ- (Fc␥RIIIa, CD16) (13–16) and the presence in most individuals of ent lymphocyte subsets was limited because the separation tech- cells with the CD3ϩCD16ϩ phenotype was confirmed (14, 17, 18). nique relied on density-gradient centrifugation of the subpopula- These cells usually comprise Ͻ2% of total PBL, with rare ex- tions of lymphocytes that form rosettes with erythrocytes coated ceptions (18). In addition, it was specified that human T lym- with rabbit IgG or IgM. Using this technique, the so-called “T␥” phocytes expressing the ␥␦ TCR can express CD16 and mediate and “T” populations were found to display different morpholog- ADCC (19–21). In contrast, the link between the expression of ical, histochemical, and functional characteristics (7, 8). In partic- CD16 and ␣ TCR on T lymphocytes has remained unclear. ular, T␥ cells have been shown to mediate ADCC (9). Later on, Although cells from large granular lymphocyte leukemia have when mAb targeting lymphoid populations became available, it been shown to coexpress CD16 and an ␣ TCR (22, 23), the became apparent that the T population was similar to the unfrac- existence of “normal” T cells harboring this phenotype has been suggested in only rare instances (24–27) and these cells have not been characterized. The present study was initiated to di- *Institut National de la Sante´et de la Recherche Me´dicale, Unite´601, Nantes, France; †l’Universite´Nantes, Nantes, France; and ‡Laboratoire d’He´matologie, Centre Hos- rectly assess the proportion, the phenotype, and the functional ϩ pitalier de l’Universite´Nantes, Nantes, France capabilities of CD16 ␣ TCR T cells. We demonstrate that ϩ Received for publication September 14, 2007. Accepted for publication February CD16 ␣ TCR T cells, which are present in small numbers in 5, 2008. all individuals 1) belong to the memory effector subset, 2) are The costs of publication of this article were defrayed in part by the payment of page perforin positive, 3) are capable of mediating ADCC immedi- charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. ately ex vivo, 4) increase proportionately in vivo during lym- 1 B.C. was supported by Association Francaise Contre les Myopathies. phocytosis, and 5) appear in vitro in all cultures of EBV-spe- 2 Address correspondence and reprint requests to Dr. Henri Vie´, Institut National de cific CTL. Consequently, because of their ADCC potential and la Sante´et de la Recherche Me´dicale, Unite´601, 9 Quai Moncousu, 44093 Nantes their presence among virus-specific T cells during a specific T cedex, France. E-mail address: [email protected] cell response, amplification of these CD16ϩ ␣ TCR effector 3 Abbreviations used in this paper: ADCC, Ab-dependent cellular cytotoxicity; memory T lymphocytes during a regular immune response may BLCL, B lymphoblastoid cell line; LGL, large granular lymphocyte. be involved in beneficial or harmful consequences depending on Copyright © 2008 by The American Association of Immunologists, Inc. 0022-1767/08/$2.00 the presence of pathogen- or tissue-specific Abs. www.jimmunol.org 5328 ADCC BY Fc␥RIIIa ␣ TCR T LYMPHOCYTES Materials and Methods trol. For ADCC assays, the indicated mAb was incubated with target cells Samples and cell lines for 20 min before addition of effector cells. After a 4-h incubation at 37°C, 25 l of supernatant were removed from each well, mixed with 100 lof PBMCs were prepared by Ficoll (PAA Laboratories) gradient centrifuga- scintillation fluid, and 51Cr activity was counted in a scintillation counter. tion of blood obtained from adult volunteers or patients with hyperlym- Each test was performed in triplicate. The results are expressed as the phocytosis that were recruited nonselectively from the Department of In- percentage of lysis, which is calculated according to the following equa- fectious Diseases (Centre Hospitalier de l’Universite´de Nantes, Nantes, tion: (experimental release Ϫ spontaneous release)/(maximal release Ϫ France). All individuals gave informed consent. EBV B lymphoblastoid spontaneous release) ϫ 100, where experimental release represents the cell lines (BLCL) were derived from PBMCs by in vitro infection using mean cpm for the target cells in the presence of effector cells, spontaneous EBV-containing culture supernatant from the Marmoset B95.8 cell line release represents the mean cpm for target cells incubated without effector purchased from the American Type Culture Collection in the presence of cells, and maximal release represents the mean cpm for target cells incu- 1 g/ml cyclosporine A. The K562 cell line was cultured in complete bated with 1% Triton X-100. For the assessment of ex vivo ADCC by ϩ medium consisting of RPMI 1640 (Sigma-Aldrich), 10% heat-inactivated CD16 ␣ T lymphocytes, cells positively sorted by the 3G8-PC5 Ab FCS, 2 mM glutamine (Sigma-Aldrich), 100 U/ml penicillin, and 10 g/ml were washed and incubated overnight in complete medium (without any streptomycin (Sigma-Aldrich). cytokines) before the functional ADCC assay. mAbs and flow cytometric analysis Statistical analysis The following mAbs and their isotype controls were used: anti-␣-FITC Differences between subjects with hyperlymphocytosis and healthy con- (BMA031; Serotec), anti-CCR7-PE (150503; RD Systems), anti-CD28-PE trols were analyzed using the t test. A p value of Ͻ0.05 was considered (L293), and anti-perforin-PE (␦69) (BD Biosciences).