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Control of Autoimmune Diabetes in NOD Mice by GAD Expression or Suppression in Beta Cells

Overview. Antisense GAD gene. Beta cell specific autoimmunity. Control of Autoimmune Diabetes in NOD Mice by GAD Expression or Suppression in Beta Cells . Conclusion. By Nicole Clarke. What is Type-1 Diabetes ?.

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Control of Autoimmune Diabetes in NOD Mice by GAD Expression or Suppression in Beta Cells

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  1. Overview Antisense GAD gene Beta cell specific autoimmunity Control of Autoimmune Diabetes in NOD Mice by GAD Expression or Suppression in Beta Cells Conclusion By Nicole Clarke

  2. What is Type-1 Diabetes ? • Type one Diabetes is also referred to as Insulin Dependant Diabetes Mellitus. It usually occurs in people under 30 years of age but may also appear at any age.

  3. GAD (Glutamic Acid Decarboxylase ) • Glutamic acid decarboxylase (GAD65) is an enzyme that is produced primarily by pancreatic islet cells.

  4. Is GAD Expression Required for Diabetes? Yes • Rational: Gad expression is necessary for diabetes • Data: Generation of a transgenic mouse that expresses antisense GAD. • Conclusion: suppression of Beta cell GAD expression was achieved by producing transgenic NOD mice with an antisense GAD transgene (RNA).

  5. Expression of antisense GAD gene at cDNA level Six lines of antisense GAD 65.67 transgenic NOD mice were established.

  6. Expression of Antisense GAD gene at the RNA level • Rational: Expression of antisense GAD gene at the RNA level • Data: viewing the transgenic (-) low, medium, and high expression of GAD • Conclusion: High antisense animals thus have antisense GAD expression of antisense GAD mRNA

  7. Expression of antisense GAD Gene at the Protein Level • Rational: Expression of antisense GAD gene at the protein level • Data: Western blot analysis showed that no GAD protein was produced in H-AS-GAD animals. • Conclusion: successful production of transgenic animal and high expression of antisense GAD meant low GAD protein

  8. Immunohistochemistry Stain of Pancreatic Cells • Rational: viewing of immunohisto-chemical stain of pancreatic cells of animals. • Data: viewing of anti-GAD, Anti-Insulin, and HE • Conclusion:A high antisense GAD mice has no GAD or DNA expressed

  9. Is GAD expression in the -cells required for the development of autoimmune diabetes in NOD mice? • Data: out of 15 animals, none of the H-AS GAD mice developed diabetes by 40 weeks of age. • Conclusion: H-AS GAD animals do not get diabetes when compared with the Tg(-).

  10. Do B-cells specific Suppression of GAD Expression Affects -cell specific autoimmunity? • Data: Islet cells are E and F while G and H are salivary cells • Conclusion: Autoimmunity will not be affected in other tissues but only in H-AS GAD NOD mice

  11. Pancreatic islets of transgenic (GAD) and transgene-negative lines severe lymphocytic infiltration Healthy Cell

  12. Do Spleen cells produce Diabetogenic T lymphocytes? • Data: Diabetic T cells of NOD SCID mice • Conclusion: There was no induction of diabetogenic T cell response by the NOD SCID mice.

  13. Splenic T- cell proliferative response to islet antigens • Data: Todetermine the proliferation of Splenic T-cells in response to islet antigen exposure thymidine was used • Conclusion: resistance of GAD transgenic NOD islet is a specific effect, because env suppressed islets were not resistant to the cytotoxic effect of diabetogenic T-cells

  14. Are the GAD suppressed islets prone to attack by diabetogenic T-cells ? • all recipients developed diabetes upon receipt of the GAD expressing islets.

  15. Findings • Yoon et al. have shown that GAD suppression via transgenic mice is specifically responsible for preventing diabetes.   • In addition, CD4+ and CD8+ T cells cannot act without GAD • GAD expression is essential for the induction of diabetogenic T cells

  16. References • CONTROL OF AUTOIMMUNE DIABETES IN NOD MICE BY GAD EXPRESSION OR SUPPRESSION IN [small beta, Greek] CELLS; Volume 284(5417) pp11183-1187; 14 May 1999 Yoon, Ji-Won; Yoon, Chang-Soon; Lim,Hye-Won; Huang, Qi Quan; Kand, Yupp; Pyun, Kwang Ho; Hirasawa, Kensuke; Sherwin, Robert S.;Jun, Hee-Sook. • Picture on slide 2 • Picture on slide 14 www.coolfun.com/mice • Movie on slide 15 www.coolfun.com/mice.mov • Dr. Peter Lin

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