Proc Natl Acad Sci U S A 100(2): 628-632

Contribution of Fas to diabetes development

^{The Jackson Laboratory, 600 Main Street, Bar Harbor, ME 04609; and}^{Section of Immunobiology, Yale University School of Medicine, and Howard Hughes Medical Institute, 310 Cedar Street, New Haven, CT 06510}

^{Present address: Juvenile Diabetes Research Foundation/Wellcome Trust Diabetes and Inflammation Laboratory, Cambridge Institute for Medical Research, Addenbrooke's Hospital, Cambridge CB2 2XY, United Kingdom.}

^{To whom correspondence should be addressed. E-mail:}gro.xaj@cva.

Contributed by Richard A. Flavell

Accepted 2002 Dec 4.

Abstract

Fas (Tnfrsf6, Apo-1, CD95) is a death receptor involved in apoptosis induced in many cell types. Fas have been shown to be expressed by insulin-producing beta cells in mice and humans. However, the importance of Fas in the development of autoimmune diabetes remains controversial. To further evaluate the importance of Fas in pathogenesis of diabetes, we generated NOD mice (nonobese diabetic mice developing spontaneous autoimmune diabetes) with beta cell-specific expression of a dominant-negative point mutation in a death domain of Fas, known as lpr^{or Fas}^{. Spontaneous diabetes was significantly delayed in NOD mice expressing Fas}^{, and the effect depended on the expression level of the transgene. However, Fas}^{-bearing mice were still sensitive to diabetes transferred by splenocytes from overtly diabetic NOD mice. At the same time, Fas}^{expression did neutralize the accelerating effect of transgenic Fas-ligand expressed by the same beta cells. Thus, both Fas-dependent and -independent mechanisms are involved in beta cell destruction, but interference with the Fas pathway early in disease development may retard or prevent diabetes progression.}

Abstract

In autoimmune diabetes, pancreatic beta cells are destroyed by effector lymphocytes, leading to the loss of insulin production and hyperglycemia (1, 2). Both CD8^{and CD4}^{T cells contribute to the process (}3), and several mechanisms of beta cell death are most likely involved (reviewed in refs. 4 and 5). We and others (6–8) found that triggering of Fas (Apo-1 and CD95) expressed on beta cells leads to their death and, subsequently, to diabetes. In our transgenic model (6), Fas ligand (FasL) was expressed in the beta cells of nonobese diabetic (NOD) mice. Although the original purpose of these experiments was to protect islet cells against autoaggressive T cells, mice that expressed FasL under the control of rat insulin promoter (RIP-FasL) (6) or human insulin promoter (9) demonstrated acceleration of both spontaneous and transferred diabetes. To explain this acceleration, a model has been suggested that predicts that transferred lymphocytes or their products induce Fas expression on beta cells, making them more susceptible to FasL-mediated killing. Normally, FasL involved in beta cell killing is expressed by effector T cells (10, 11). However, in the case of RIP-FasL transgenics, the ectopic expression of FasL accelerates the killing, most likely by a fratricide mechanism.

To further study the involvement of Fas in the destruction of insulin-producing cells, we used mice with a natural Fas mutation −lpr (12). We also produced NOD mice with dominant-negative mutations of Fas-lpr^{called Fas}^{here (}13), which expressed the transgene selectively in the islets of Langerhans.

Acknowledgments

We thank Jeffrey Bedrozian for technical assistance, Dr. Susan F. Wong (University of Bristol, Bristol, U.K.) for the gift of IS-CD8^{cells and helpful discussions, Dr. S. Nagata (Osaka University Medical School, Osaka) for the kind gift of Fas cDNA, and Drs. L. Matis and Y. Wang (Alexion Pharmaceuticals, New Haven, CT) for the gift of NOD-lpr mice. This work was supported by National Institutes of Health Grant IDDK53561, Juvenile Diabetes Research Foundation International Grants JDRF-166 and JDRF-546 (to A.V.C.), and a Postdoctoral Fellowship from the Juvenile Diabetes Research Foundation International (to A.Y.S.). R.A.F. is an Investigator of the Howard Hughes Medical Institute.}

Acknowledgments

Abbreviations

FasL	Fas ligand
IS-CD8^cells	insulin-specific CD8^{T cells}
LSD	least significant difference
NOD	nonobese diabetic
RIP	rat insulin promoter

Abbreviations

References

1. Tisch R, McDevitt H O. Cell. 1996;85:291–297.[PubMed]
2. Delovitch T L, Singh B. Immunity. 1997;7:727–738.[PubMed]
3. Wong F S, Janeway C A., Jr J Autoimmun. 1999;13:290–295.[PubMed]
4. Chervonsky A V. Curr Opin Immunol. 1999;11:684–688.[PubMed]
5. Eizirik D L, Mandrup-Poulsen T. Diabetologia. 2001;44:2115–2133.[PubMed]
6. Chervonsky A V, Wang Y, Wong F S, Visintin I, Flavell R A, Janeway C A, Jr, Matis L A. Cell. 1997;89:17–24.[PubMed]
7. Nakayama M, Nagata M, Yasuda H, Arisawa K, Kotani R, Yamada K, Chowdhury S A, Chakrabarty S, Jin Z Z, Yagita H, et al Diabetes. 2002;51:1391–1397.[PubMed][Google Scholar]
8. Amrani A, Verdaguer J, Thiessen S, Bou S, Santamaria P. J Clin Invest. 2000;105:459–468.
9. Petrovsky N, Silva D, Socha L, Slattery R, Charlton B. Ann NY Acad Sci. 2002;958:204–208.[PubMed]
10. Amrani A, Verdaguer J, Anderson B, Utsugi T, Bou S, Santamaria P. J Clin Invest. 1999;103:1201–1209.
11. Wong F S, Visintin I, Wen L, Flavell R A, Janeway C A. J Exp Med. 1996;183:67–76.
12. Watanabe-Fukunaga R, Brannan C I, Copeland N G, Jenkins N A, Nagata S. Nature. 1992;356:314–317.[PubMed]
13. Matsuzawa A, Moriyama T, Kaneko T, Tanaka M, Kimura M, Ikeda H, Katagiri T. J Exp Med. 1990;171:519–531.
14. Lo D, Burkly L C, Widera G, Cowing C, Flavell R A, Palmiter R D, Brinster R L. Cell. 1988;8:159–168.[PubMed]
15. Guerder S, Picarella D E, Linsley P S, Flavell R A. Proc Natl Acad Sci USA. 1994;91:5138–5142.
16. Chirgwin J M, Przybyla A E, MacDonald R J, Rutter W J. Biochemistry. 1979;18:5294–5299.[PubMed]
17. Siegling A, Lehmann M, Platzer C, Emmrich F, Volk H D. J Immunol Methods. 1994;177:23–28.[PubMed]
18. Wong F S, Karttunen J, Dumont C, Wen L, Visintin I, Pilip I M, Shastri N, Pamer E G, Janeway C A. Nat Med. 1999;9:1026–1031.[PubMed]
19. Savinov A Y, Wong F S, Chervonsky A V. J Immunol. 2001;167:6637–6643.[PubMed]
20. Yabuki A, Suzuki S, Matsumot M, Kurohmaru M, Hayashi Y, Nishinakagawa H. J Vet Med Sci. 2001;63:1339–1342.[PubMed]
21. Venables W N, Ripley B D Modern Applied Statistics with S-PLUS. 3rd Ed. New York: Springer; 1999. [PubMed][Google Scholar]
22. Kang S M, Schneider D B, Lin Z, Hanahan D, Dichek D A, Stock P G, Baekkeskov S. Nat Med. 1997;3:738–743.[PubMed]
23. Allison J, Georgiou H M, Strasser A, Vaux D L. Proc Natl Acad Sci USA. 1997;94:3943–3947.
24. Nelson D P, Setser E, Hall D G, Schwartz S M, Hewitt T, Klevitsky R, Osinska H, Bellgrau D, Duke R C, Robbins J. J Clin Invest. 2000;105:1199–1208.
25. Shudo K, Kinoshita K, Imamura R, Fan H, Hasumoto K, Tanaka M, Nagata S, Suda T. Eur J Immunol. 2001;31:2504–2511.[PubMed]
26. Lewinsohn D M, Bargatze R F, Butcher E C. J Immunol. 1987;138:4313–4321.[PubMed]
27. Lagasse E, Weissman I L. J Immunol Methods. 1996;197:139–150.[PubMed]
28. Itoh N, Imagawa A, Hanafusa T, Waguri M, Yamamoto K, Iwahashi H, Moriwaki M, Nakajima H, Miyagawa J, Namba M, et al J Exp Med. 1997;186:613–618.[Google Scholar]
29. Kim S, Kim K A, Hwang D Y, Lee T H, Kayagaki N, Yagita H, Lee M S. J Immunol. 2000;164:2931–2936.[PubMed]
30. Chu J L, Ramos P, Rosendorff A, Nikolic-Zugic J, Lacy E, Matsuzawa A, Elkon K B. J Exp Med. 1995;181:393–398.
31. Allison J, Strasser A. Proc Natl Acad Sci USA. 1998;95:13818–13822.
32. Su X, Hu Q, Kristan J M, Costa C, Shen Y, Gero D, Matis L A, Wang Y. J Immunol. 2000;164:2523–2532.[PubMed]
33. Huang B, Eberstadt M, Olejniczak E T, Meadows R P, Fesik S W. Nature. 1996;384:638–641.[PubMed]
34. Itoh N, Nagata S. J Biol Chem. 1993;268:10932–10937.[PubMed]
35. Boldin M P, Varfolomeev E E, Pancer Z, Mett I L, Camonis J H, Wallach D. J Biol Chem. 1995;270:7795–7798.[PubMed]
36. Ogata Y, Kimura M, Shimada K, Wakabayashi T, Onoda H, Katagiri T, Matsuzawa A. Cell Immunol. 1993;148:91–102.[PubMed]