In Situ β Cell Death Promotes Priming of Diabetogenic CD8 T Lymphocytes Yiqun Zhang, Bronwyn O'Brien, Jacqueline Trudeau, Rusung Tan, Pere Santamaria and Jan P. Dutz This information is current as of September 26, 2021. J Immunol 2002; 168:1466-1472; ; doi: 10.4049/jimmunol.168.3.1466 http://www.jimmunol.org/content/168/3/1466 Downloaded from References This article cites 64 articles, 39 of which you can access for free at: http://www.jimmunol.org/content/168/3/1466.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 26, 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. In Situ ␤ Cell Death Promotes Priming of Diabetogenic CD8 T Lymphocytes1 Yiqun Zhang,* Bronwyn O’Brien,‡ Jacqueline Trudeau,†‡ Rusung Tan,† Pere Santamaria,§ Jan P. Dutz2* CTLs are important mediators of pancreatic ␤ cell destruction in the nonobese diabetic mouse model of type 1 diabetes. Cross- presentation of Ag is one means of priming CTLs. The death of Ag-bearing cells has been implicated in facilitating this mode of priming. The role of ␤ cell death in facilitating the onset of spontaneous autoimmune diabetes is unknown. Here, we used an adoptive transfer system to determine the time course of islet-derived Ag presentation to naive ␤ cell-specific CD8 T cells in nonobese diabetic mice and to test the hypothesis that ␤ cell death enhances the presentation of ␤ cell autoantigen. We have determined that ␤ cell death enhances autoantigen presentation. Priming of diabetogenic CD8 T cells in the pancreatic lymph nodes was negligible before 4 wk, progressively increased until 8 wk of age, and was not influenced by gender. Administration of Downloaded from multiple low doses of the ␤ cell toxin streptozotocin augmented in situ ␤ cell apoptosis and accelerated the onset and magnitude of autoantigen presentation to naive CD8 T cells. Increasing doses of streptozotocin resulted in both increased pancreatic ␤ cell death and significantly enhanced T cell priming. These results indicate that in situ ␤ cell death facilitates autoantigen-specific CD8 T cell priming and can contribute to both the initiation and the ongoing amplification of an autoimmune response. The Journal of Immunology, 2002, 168: 1466–1472. http://www.jimmunol.org/ ype 1 diabetes mellitus (DM)3 is an autoimmune disorder transfer of naive CD4 T cells specific for a pancreatic ␤ cell Ag, in which the pancreatic ␤ cells are destroyed by the cel- it has been demonstrated that these cells can be primed in the T lular immune system (1, 2). The nonobese diabetic pancreatic lymph nodes as early as, but not before, 14 days of age (NOD) mouse is a useful and well-studied animal model for hu- (14). Similarly, CD8 T cells responding to transgenically overex- man type 1 DM. Adoptive T cell transfer studies using T cells from pressed Ags in the pancreas cannot be primed in mice of 5–10 days prediabetic NOD mice have demonstrated that transfer of diabetes of age (14, 15). The timing and mechanism by which CD8 T cells into immunodeficient mice requires both CD4 and CD8 T cells are primed to pancreatic ␤ cell Ag in spontaneous autoimmunity (3–6). CD8 T cells are known to participate in pancreatic ␤ cell remains to be determined. Cross-presentation of Ag, where tissue- destruction (7–9). The importance of CD8 T cells in diabetogen- specific Ag is processed by APCs and presented to T cells via class by guest on September 26, 2021 esis is underscored by the fact that neither DM nor insulitis de- I MHC, is one means of priming CD8 T cells (16). Cross-presen- velop in NOD mice deficient in MHC class I expression and CD8 tation of cellular Ags expressed by tumors (17), viruses (18), and T cells (10, 11). Recent data suggest that CD8 T cells play a piv- self (19), as well as of soluble Ags, has been described (20). Cell otal role in the earliest initiative phases of autoimmune pancreatic death, specifically by apoptosis, can provide an excellent source of ␤ cell destruction (12, 13). cellular Ag for cross-presentation (21, 22). Cell death, by either The activation of naive (resting and previously unactivated) T apoptosis (23) or necrosis (24), can also provide cell-derived sig- cells requires activation of the TCR and the simultaneous delivery nals that activate APCs and promote immune responses. There- of a costimulatory signal by a specialized APC . Using adoptive fore, we hypothesized that ␤ cell death could promote the initiation and/or progression of spontaneous autoimmune diabetes. The aims of the present study were first to delineate the timing Departments of *Medicine and †Pathology and Laboratory Medicine, British Colum- bia Research Institute of Children and Women’s Health, University of British Co- of CD8 T cell priming to pancreatic ␤ cell-associated Ag in NOD lumbia, Vancouver, British Columbia, Canada; ‡Department of Kinesiology, Simon mice and, second, to determine whether ␤ cell death can promote Fraser University, Burnaby, British Columbia, Canada; and §Department of Micro- biology and Infectious Diseases, Faculty of Medicine, University of Calgary, Calgary, the priming of CD8 T cells to a diabetogenic self-Ag. We used an Alberta, Canada adoptive transfer system using CD8 T cells derived from a mouse Received for publication September 19, 2001. Accepted for publication November line that transgenically expresses a TCR of a diabetogenic T cell 13, 2001. clone (8). This clone uses a TCR rearrangement expressed by a The costs of publication of this article were defrayed in part by the payment of page preponderance of CD8 T cells found in early insulitic lesions of charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. NOD mice (13). The results of our investigations show that prim- ing of diabetogenic CD8 T cells occurs in the pancreatic lymph 1 This work was supported by a grant from the Canadian Institutes of Health Re- search/Juvenile Diabetes Research Foundation International Program II (to J.P.D.), nodes and is undetectable before 3–4 wk of age and then progres- and grants from the Canadian Institutes of Health Research and Canadian Diabetes sively increases with age to reach maximal levels at 8 wk of age. Association (to P.S.). J.P.D. is a Junior Scholar of the Arthritis Society. P.S. is a Senior Scholar for the Alberta Heritage Foundation for Medical Research. All authors An increased incidence of apoptosis in the pancreas after the ad- are members of the Beta Cell Apoptosis Network. ministration of low doses of the ␤ cell toxin streptozotocin (STZ) 2 Address correspondence and reprint requests to Dr. Jan P. Dutz at the current ad- augmented CD8 T cell priming and allowed it to occur at ages dress: The Skin Care Center, 835 West Tenth Avenue, Vancouver, British Columbia when it does not occur spontaneously. These results suggest that ␤ V5Z 4E2, Canada. E-mail address: [email protected] cell death precedes the priming of T cells in autoimmune diabetes 3 Abbreviations used in this paper: DM, diabetes mellitus; NOD, nonobese diabetic; ␤ STZ, streptozotocin; CM, complete medium; DC, dendritic cell; PARP, poly(A)DP and implicate in situ cell death in the facilitation of autoantigen- ribose polymerase. specific CD8 T cell priming in autoimmunity. Copyright © 2002 by The American Association of Immunologists 0022-1767/02/$02.00 The Journal of Immunology 1467 Materials and Methods achieved with 3,3Ј-diaminobenzidine tetrahydrochloride as the substrate. Mice and cell lines To provide a negative control for the TUNEL, TdT was omitted from the labeling mix. The ␤ cell origin of the apoptosis was verified by double NOD mice were obtained from Taconic Farms (Germantown, NY) and staining for insulin using guinea pig anti-porcine insulin Ab (DAKO, Mis- bred in a specific pathogen-free environment. The 8.3-NOD mice, express- sissauga, Ontario, Canada), a biotinylated anti-guinea pig IgG as secondary ing the rearranged TCR genes of the diabetogenic CTL clone NY8.3, have Ab (Vector Laboratories, Burlington, Ontario, Canada), and the chromogen been previously described (8) and were bred at the University of Calgary New Fuschin (DAKO). Cells with brown nuclei (TUNEL positive) and (Calgary, Alberta, Canada). C57BL/6 mice were obtained from The Jack- pink cytoplasm (insulin positive) were identified as apoptotic ␤ cells. Ap- son Laboratory (Bar Harbor, ME). All animal experiments were approved optosis was quantified in all ␤ cells present in stained sections from each by the Animal Care Committee of the University of British Columbia. EL4 animal, each section being separated from the next by at least 40 ␮m. thymoma and E.G7-OVA cells (EL4 cells transfected to express OVA) Approximately 1000 ␤ cells were counted for each animal by a single (20) were obtained from the American Type Culture Collection (Manassas, observer, blinded to the treatment status of the animal. The number of VA) and cultured in complete medium (CM) consisting of RMPI 1640 apoptotic ␤ cells present was expressed as a percentage of the total number medium (Invitrogen, Burlington, Ontario, Canada) supplemented with of ␤ cells observed.