The Analysis of CD83 Expression on Human Immune Cells Identifies a Unique CD83 + -Activated T Cell Population This information is current as Xinsheng Ju, Pablo A. Silveira, Wei-Hsun Hsu, Zehra of October 1, 2021. Elgundi, Renz Alingcastre, Nirupama D. Verma, Phillip D. Fromm, Jennifer L. Hsu, Christian Bryant, Ziduo Li, Fiona Kupresanin, Tsun-Ho Lo, Candice Clarke, Kenneth Lee, Helen McGuire, Barbara Fazekas de St. Groth, Stephen R. Larsen, John Gibson, Kenneth F. Bradstock, Georgina J. Clark and Derek N. J. Hart Downloaded from J Immunol 2016; 197:4613-4625; Prepublished online 11 November 2016; doi: 10.4049/jimmunol.1600339 http://www.jimmunol.org/content/197/12/4613 http://www.jimmunol.org/ Supplementary http://www.jimmunol.org/content/suppl/2016/11/11/jimmunol.160033 Material 9.DCSupplemental References This article cites 50 articles, 24 of which you can access for free at: http://www.jimmunol.org/content/197/12/4613.full#ref-list-1 by guest on October 1, 2021 Why The JI? 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The Journal of Immunology The Analysis of CD83 Expression on Human Immune Cells Identifies a Unique CD83+-Activated T Cell Population Xinsheng Ju,* Pablo A. Silveira,*,† Wei-Hsun Hsu,*,† Zehra Elgundi,* Renz Alingcastre,* Nirupama D. Verma,* Phillip D. Fromm,*,† Jennifer L. Hsu,*,‡ Christian Bryant,*,‡ Ziduo Li,*,† Fiona Kupresanin,* Tsun-Ho Lo,*,† Candice Clarke,x Kenneth Lee,†,x Helen McGuire,{ Barbara Fazekas de St. Groth,{ Stephen R. Larsen,†,‡ John Gibson,†,‡ Kenneth F. Bradstock,*,† Georgina J. Clark,*,† and Derek N. J. Hart*,† CD83 is a member of the Ig gene superfamily, first identified in activated lymphocytes. Since then, CD83 has become an important marker for defining activated human dendritic cells (DC). Several potential CD83 mRNA isoforms have been described, including a soluble form detected in human serum, which may have an immunosuppressive function. To further understand the biology of CD83, we examined its expression in different human immune cell types before and after activation using a panel of mouse Downloaded from and human anti-human CD83 mAb. The mouse anti-human CD83 mAbs, HB15a and HB15e, and the human anti-human CD83 mAb, 3C12C, were selected to examine cytoplasmic and surface CD83 expression, based on their different binding charac- teristics. Glycosylation of CD83, the CD83 mRNA isoforms, and soluble CD83 released differed among blood DC, monocytes, and monocyte-derived DC, and other immune cell types. A small T cell population expressing surface CD83 was identified upon T cell stimulation and during allogeneic MLR. This subpopulation appeared specifically during viral Ag challenge. We did not observe human CD83 on unstimulated human natural regulatory T cells (Treg), in contrast to reports describing expression of CD83 on http://www.jimmunol.org/ mouse Treg. CD83 expression was increased on CD4+, CD8+ T, and Treg cells in association with clinical acute graft-versus-host disease in allogeneic hematopoietic cell transplant recipients. The differential expression and function of CD83 on human immune cells reveal potential new roles for this molecule as a target of therapeutic manipulation in transplantation, inflammation, and autoimmune diseases. The Journal of Immunology, 2016, 197: 4613–4625. D83 is a member of the Ig superfamily of receptors, human CD83 (hCD83) are 63% identical, but the mouse protein consisting of a single variable-like Ig domain, a trans- lacks a 10-aa portion of the extracellular Ig domain. How this by guest on October 1, 2021 C membrane domain, and C-terminal cytoplasmic domain influences the expression or function of mCD83 is unknown, but (1). The amino acid sequences of mouse CD83 (mCD83) and differences between the species might be expected (1, 2). hCD83 is expressed both as membrane-bound and soluble CD83 (sCD83) protein (3), but data on naturally derived sCD83 in mice are lacking. *ANZAC Research Institute, Concord Repatriation General Hospital, Sydney, New Likewise, CD83 mRNA splice variants are present in human cells, South Wales 2139, Australia; †Sydney Medical School, University of Sydney, Syd- ney, New South Wales 2006, Australia; ‡Institute of Haematology, Royal Prince but the status of their translation remains unknown and similar splice Alfred Hospital, University of Sydney, Sydney, New South Wales 2050, Australia; variants were not very evident in mice (4, 5). xAnatomical Pathology Department, Concord Repatriation General Hospital, Sydney, New South Wales 2139, Australia; and {Centenary Institute, Royal Prince Alfred CD83 on thymic epithelial cells enables mouse T cells to develop + Hospital, Sydney, New South Wales 2050, Australia from double-negative thymocytes to mature naive CD4 T cells ORCIDs: 0000-0001-9755-9908 (X.J.); 0000-0001-5074-8824 (P.A.S.); 0000- (6), and its engagement on mouse B cells modulates their function 0002-6902-4719 (W.-H.H.); 0000-0002-4456-9368 (R.A.); 0000-0001-7047- (7). Cell surface CD83 is induced on mouse activated dendritic 1113 (P.D.F.); 0000-0001-9920-0720 (J.L.H.); 0000-0002-4173-6394 (Z.L.); 0000-0003-0229-6004 (T.-H.L.); 0000-0001-8700-5581 (K.L.); 0000-0003- cells (DC) (8) and macrophages (9). Recently, it was described on 2047-6543 (H.M.); 0000-0001-6817-9690 (B.F.d.S.G.); 0000-0001-7894- unactivated mouse regulatory T cells (Treg) (10, 11). Surface 232X (G.J.C.); 0000-0003-2620-1612 (D.N.J.H.). CD83 expression is upregulated upon human DC activation (1) Received for publication March 2, 2016. Accepted for publication October 10, 2016. and is commonly used to identify DC maturation/activation. In This work was supported by Australian Government National Health Medical Research addition, it is induced on activated human B cells (12, 13), mono- Council Program Grant 543727 (to D.N.J.H. and K.F.B.) and Cancer Institute New cytes (14), macrophages (14), neutrophils (15), and a regulatory South Wales Translational Program Grant 11/TPG/3-02 (to D.N.J.H. and K.F.B.). The Cooperative Research Centre for Biomarker Translation also provided research support. subset of NK cells (16). We and others described CD83 on activated Address correspondence and reprint requests to Prof. Derek N.J. Hart, ANZAC human T cells (6, 10, 17), but its expression on specific T cell Research Institute, Concord Repatriation General Hospital, Gate 3, Hospital Road, subsets has not been well scrutinized (18). Sydney, NSW 2139, Australia. E-mail address: [email protected] We previously identified sCD83 in human serum, described its The online version of this article contains supplemental material. release from activated DC and B lymphocytes (19), and suggested Abbreviations used in this article: AF, AlexaFluor; alloHCT, allogenic hematopoietic that sCD83 was produced by the shedding of cell surface CD83 cell transplant; AlloPC, allophycocyanin; BV, Brilliant Violet; CEF, PepMix CMV, EBV, influenza A; Cy, cyanine; DC, dendritic cell; GVHD, graft-versus-host disease; molecules, although translation from a specific CD83 splice var- hCD83, human CD83; mCD83, mouse CD83; Mo-DC, monocyte-derived DC; pDC, iant remains a formal possibility (3). Recombinant sCD83-Fc plasmacytoid DC; sCD83, soluble CD83; TGN, trans–Golgi network 46; Treg, reg- fusion proteins have been shown to inhibit T cell proliferation ulatory T cell. (4, 20) and DC maturation (21), suppress chronic inflammatory Copyright Ó 2016 by The American Association of Immunologists, Inc. 0022-1767/16/$30.00 diseases (22), and prolong allogeneic corneal and renal transplant www.jimmunol.org/cgi/doi/10.4049/jimmunol.1600339 4614 CD83 EXPRESSION ON HUMAN IMMUNE CELLS survival in murine models (23, 24). Increased serum levels of lated by MACS positive selection using CD14 microbeads (Miltenyi sCD83 have been reported in patients with hematopoietic malig- Biotec). T cells were isolated with an EasySep human T cell isolation kit nancies and rheumatoid arthritis (25, 26). (STEMCELL Technologies). The CD304 (BDCA-4) microbead kit (Mil- tenyi Biotec) was used to deplete plasmacytoid DC (pDC) from PBMC. In Investigations in CD83-deficient, mutant, and transgenic re- the experiments indicated, DC, monocytes, B cells, NK cells, or T cells porter mice (6, 7, 27) have suggested both direct and indirect roles were sorted using an Influx Cell Sorter System (BD Biosciences), and the for CD83 in T cell development and function. CD83-deficient gating strategy is shown (Supplemental Fig. 1). + mice have reduced numbers of CD4 T cells, attributed to de- Abs fects in thymic T cell selection secondary to altered Ag presen- tation by thymic epithelial cells (6). The CD83 transmembrane The anti-human Ab conjugates used included the following: CD3 (SP34-2; Brilliant Violet [BV]450 or AlexaFluor [AF]700), CD4 (L200; PerCP- region