CD30 Expression Identifies the Predominant Proliferating T Lymphocyte Population in Human Alloimmune Responses This information is current as Keith W. Chan, Corwyn D. Hopke, Sheri M. Krams and of October 5, 2021. Olivia M. Martinez J Immunol 2002; 169:1784-1791; ; doi: 10.4049/jimmunol.169.4.1784 http://www.jimmunol.org/content/169/4/1784 Downloaded from References This article cites 39 articles, 23 of which you can access for free at: http://www.jimmunol.org/content/169/4/1784.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 *average by guest on October 5, 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 © 2002 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology CD30 Expression Identifies the Predominant Proliferating T Lymphocyte Population in Human Alloimmune Responses1 Keith W. Chan,* Corwyn D. Hopke,† Sheri M. Krams,*† and Olivia M. Martinez2*† CD30 is an inducible member of the TNFR superfamily that is expressed on activated T and B cells and some lymphoid malig- nancies. We have previously shown that human CD30؉ T cells elicited with allogeneic APC are a major source of IFN-␥ and IL-5 production. In the present study we have used alloantigen, as well as anti-CD3 plus anti-CD28 mAb stimulation, to further -characterize human CD30؉ T cells with respect to function and the expression of other activation-dependent cell surface mole cules, including the related TNFR family members OX-40 and 4-1BB (CD137). Our results indicate that human CD30؉ T cells are a subset of activated T cells that also express CD25 and CD45RO. Moreover, we observed that allogeneic APC consistently induced a greater proportion of CD30؉ cells within the activated T cell population than did stimulation with plate-bound anti-CD3 plus anti-CD28 mAb or stimulation with soluble anti-CD3 plus anti-CD28 and autologous APC. The enhanced induction of CD30 expression by alloantigen was not common to other inducible TNFR family members because anti-CD3 plus anti-CD28 mAbs were Downloaded from far more effective in inducing expression of 4-1BB and OX-40. Furthermore, CD30 expression marked the predominant prolif- erating T cell population induced by alloantigen as determined by CFSE staining and flow cytometry. These results indicate that CD30, but not 4-1BB or OX-40, is preferentially induced by alloantigen, suggesting that CD30 may be important in human alloimmune responses. The Journal of Immunology, 2002, 169: 1784–1791. D30 is a 120-kDa type 1 transmembrane glycoprotein to induce apoptosis during negative selection in the thymus (12, http://www.jimmunol.org/ and a member of the TNFR superfamily (1). CD30 was 13), although some studies fail to support a role for CD30 in this C originally identified as a molecule expressed on Hodgkin process (14). and Reed Sternberg cells in Hodgkin’s disease but has subse- Other inducible members of the TNFR family expressed on T quently been detected on certain non-Hodgkin’s malignancies as cells, including 4-1BB and OX-40, have also been shown to have well as some virally transformed cells (2). CD30 is also expressed costimulatory function. The interaction between 4-1BB (CD137) on a subset of normal human T lymphocytes following activation and 4-1BB ligand (4-1BBL), expressed on activated B cells, mac- with Ag or mitogen (3, 4). Expression of CD30 has been proposed rophages, and dendritic cells, is particularly important in CD8- to preferentially mark Th2 cells (5), although CD30 has also been mediated responses to alloantigen (15–17), viruses (15, 18), and by guest on October 5, 2021 detected on CD4ϩ Th0 and Th1 cells, indicating that CD30 is not tumors (19), although recent studies also suggest a role for 4-1BB/ restricted to Th2 cells (6, 7). We (8) and others (9) have demon- 4-1BBL interactions in CD4ϩ T cell function (20). OX-40, in con- strated that primary stimulation of human T cells with alloantigen trast, primarily promotes cytokine production by CD4ϩ T cells or anti-CD3 mAbs and IL-2 elicits a population of CD30ϩ T cells (21–23) and has been implicated in T cell function in animal mod- that is a major source of IFN-␥ and IL-5 production. els of allogeneic graft-vs-host disease (GVHD) (24), experimental Several lines of evidence indicate that CD30 acts as a costimu- allergic encephalomyelitis (25), leishmaniasis (26), inflammatory latory molecule in T cell responses. The human CD30 ligand bowel disease (27), and tumor immunity (28). 4-1BB and OX-40, (CD30L3; CD153) is expressed in the periphery on activated T like CD30, tend to be expressed relatively late following activa- cells, monocytes, B cells, neutrophils, and eosinophils. Engage- tion, suggesting these molecules may be important in sustaining T ment of CD30 by CD30Lϩ cells or agonist anti-CD30 mAbs in- cell responses after the initial CD28/B7-1 and B7-2 interactions. duces IFN-␥ production (9) and proliferation (10) in anti-CD3- Despite the observed similarities in expression and function of stimulated peripheral blood T cells, NF-␬B activation, and viral CD30, OX-40, and 4-1BB in murine systems, relatively little is expression in an HIV-infected T cell line (11), and Ca2ϩ flux in known about the expression patterns of these inducible TNFR fam- Jurkat cells (3). Signaling through CD30 has also been suggested ily members on human T lymphocytes. The purpose of the current study was to examine the relationship of CD30 expression to the expression of other inducible TNFR family members on human T *Department of Surgery and †Program in Immunology, Stanford University School of cells and to further characterize the phenotype and function of Medicine, Stanford, CA 94305 CD30ϩ T cells in alloimmune responses. Received for publication October 11, 2001. Accepted for publication June 13, 2002. The costs of publication of this article were defrayed in part by the payment of page Materials and Methods charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. Abs and reagents 1 This work was supported by National Institutes of Health Grant RO1 DK47810 (to The following Abs were purchased from DAKO (Carpinteria, CA): anti- O.M.M.). K.W.C. was supported by a Howard Hughes Medical Institute Fellowship. CD3-FITC, anti-CD3-PECy5, anti-CD4-PE, anti-CD4-PECy5, anti-CD8- 2 Address correspondence and reprint requests to Dr. Olivia M. Martinez, Stanford PECy5, anti-CD30-FITC, anti-CD25-PE, IgG1-FITC, IgG1-PE, and PE- University School of Medicine, 1201 Welch Road, Medical School Lab Surge P312, conjugated rabbit anti-mouse Ig. PE-conjugated mAbs specific for CD25, Stanford, CA 94305-5492. E-mail address: [email protected] OX-40, 4-1BB, CD27, and CD45RO, as well as IgG1-PECy5, were ob- 3 Abbreviations used in this paper: CD30L, CD30 ligand; 4-1BBL, 4-1BB ligand; tained from BD PharMingen (San Diego, CA). Anti-CD28 and anti-CD30 GVHD, graft-vs-host disease; LCL, lymphoblastoid B cell line; NHS, normal human mAbs were purchased from BD PharMingen and anti-CD3 mAb was pur- serum; P/S, penicillin-streptomycin. chased from Coulter (Miami, FL). Human rIL-2 was obtained from the Copyright © 2002 by The American Association of Immunologists, Inc. 0022-1767/02/$02.00 The Journal of Immunology 1785 Biological Resources Branch of the National Cancer Institute (Frederick, anti-CD4-PECy5, or anti-CD8-PECy5 in addition to FITC- and PE-conju- MD). Propidium iodide was purchased from Sigma-Aldrich (St. Louis, gated mAbs, the cells were further gated on the CD3-, CD4- or CD8- MO) and CFSE was obtained from Molecular Probes (Eugene, OR). positive populations to analyze T cell subsets. Cell preparation CFSE labeling and analysis of cell division by flow cytometry Lymphoblastoid B cell lines (LCL) were maintained in RPMI 1640 with Isolated PBMC were washed, counted, and resuspended at 5 ϫ 107/ml in 10% FCS and 1% penicillin-streptomycin (P/S) in a 5% CO2 atmosphere RPMI 1640 with 1% P/S. A stock solution of CFSE (5 mM) was diluted at 37°C. Blood was collected from healthy donors by venipuncture in hep- 1/10 with RPMI 1640 and 15 ␮l of the CFSE was added per milliliter of arinized tubes. PBMC were isolated from blood samples by Ficoll- cells. The cell suspension was incubated for 10 min at 37°C. The cells were Hypaque (Amersham Pharmacia Biotech, Piscataway, NJ) density centrif- then washed and cultured in RPMI 1640 supplemented with 10% NHS and ugation. PBMC were washed, counted, and cultured in RPMI 1640 with 1% P/S on plates precoated with anti-CD3 plus anti-CD28 mAbs or with 10% normal human serum (NHS; Sigma-Aldrich) and 1% P/S at 37°Cin allogeneic APC in MLR as described above. Cells were collected on days a humidified incubator with 5% CO2. T cells were isolated by negative 3–6, washed with PBS/FCS, and stained with unconjugated mouse anti- selection using the MiniMACS (Miltenyi Biotec, Auburn, CA) separation human CD30 mAbs for 20 min at 4°C.