Mechanisms of oral tolerance to insulin in offspring of rats with experimental gestational diabetes

Öz

Background: The aim of our work was to find out the mRNA gene expression level of Aire, Deaf1, Foxp3, Ctla4 and IL10 in mesenteric lymph nodes in the offspring of rats with experimental gestational diabetes and in conditions of insulin oral tolerance formation.

Material and Methods: Using molecular genetic and immunofluorescence techniques we investigated the mRNA gene expression level of Aire, Deaf1, Foxp3, Ctla4 and IL10 in the offspring of rats with experimental gestational diabetes and in conditions of insulin oral tolerance formation. To determine the level of mRNA studied gene RT-PCR was performed in real time thermocycler CFX96 ™ Real-Time PCR Detection Systems.

Results: We observed such violations of immunotolerance: AIRE gene repression, reduced mRNA levels of Deaf1 and the transcription factor Foxp3. This was accompanied by inhibition of gene expression suppressor cytokine IL-10 and negative costimulatory molecules Ctla4.

Conclusion: Oral insulin during the first 2 weeks graded these changes, causing transcriptional activation of genes AIRE, Deaf1, Foxp3, Ctla4 and Il-10.

Anahtar Kelimeler

• Experimental gestational diabetes,mesenteric lymph nodes,Aire,Deaf1,Foxp3

Kaynakça

1. 1. Metzger T, Anderson M. Control of central and peripheral tolerance by Aire. Immunol Rev 2011; 241: 89-103.
2. 2. Kojima H, Fujimiya M, Matsumura K. Extrapancreatic insulin-producing cells in multiple organs in diabetes. Proc. Natl. Acad. Sci USA 2004; 101: 2458-2463.
3. 3. Cohen J, Tewalt E, Rouhani S. Tolerogenic properties of lymphatic endothelial cells are controlled by the lymph node microenvironment. PLoS ONE 2014; 9:e87740.
4. 4. Yip L, Su L, Sheng D. Deaf1 isoforms control the expression of genes encoding peripheral tissue antigens in the pancreatic lymph nodes during type 1 diabetes. Nat. Immunol. 2009:1026-1033
5. 5. Shevach E, Thornton A. tTregs, pTregs, and iTregs: similarities and differences. Immunol Rev 2014;259:88-102.
6. 6. Wang S, Gao X, Shen G, Wang W, Li J, Zhao J. Interleukin-10 deficiency impairs regulatory T cell-derived neuropilin-1 functions and promotes Th1 and Th17 immunity. Sci Rep 2016;6:24249.
7. 7. Walker LS, Sansom DM. Confusing signals: recent progress in CTLA-4 biology. Trends Immunol 2015;36:63-70.
8. 8. Hammerschmidt SI, Ahrendt M, Bode U, Wahl B, Kremmer E, Förster R, Pabst O. Stromal mesenteric lymph node cells are essential for the generation of gut-homing T cells in vivo. J Exp Med 2008;205:2483-2490.

9. 9. Yang S, Fujikado N, Kolodin D, Benoist C, Mathis D. Immune tolerance. Regulatory T cells generated early in life play a distinct role in maintaining self-tolerance. Science 2015;348:589-594.
10. 10. Macpherson A, Smith K. Mesenteric lymph nodes at the center of immune anatomy. J Exp Med 2006;203:497-500.
11. 11. Kunkel D, Kirchhoff D, Nishikawa S, Radbruch A, Scheffold A. Visualization of peptide presentation following oral application of antigen in normal and Peyer’s patches-deficient mice. Eur J Immunol 2003;33:1292-1301.
12. 12. Spahn TW, Weiner HL, Rennert PD, Lügering N, Fontana A, Domschke W, Spahn T.W. Mesenteric lymph nodes are critical for the induction of high dose oral tolerance in the absence of Peyer’s patches. Eur J Immunol 2002;32:1109-1113.
13. 13. Huang FP, Platt N, Wykes M, Major JR, Powell TJ, Jenkins CD, MacPherson GG. A discrete subpopulation of dendritic cells transports apoptotic intestinal epithelial cells to T cell areas of mesenteric lymph nodes. J Exp Med 2000;191:435-444.
14. 14. Worbs T, Bode U, Yan S, Hoffmann MW, Hintzen G, Bernhardt G. Oral tolerance originates in the intestinal immune system and relies on antigen carriage by dendritic cells. J Exp Med 2006;203:519-527.
15. 15. Roep B, Tree T. Immune modulation in humans: implications for type 1 diabetes mellitus. Nat Rev Endocrinol 2014;10:229-242.
16. 16. Gale EA, Bingley PJ, Emmett CL, Collier T. European Nicotinamide Diabetes Intervention Trial (ENDIT): a randomised controlled trial of intervention before the onset of type 1 diabetes. Lancet 2004;363:925-31.
17. 17. Bonifacio E, Ziegler AG, Klingensmith G, Schober E, Bingley PJ, Rottenkolber M. Effects of high-dose oral insulin on immune responses in children at high risk for type 1 diabetes: the Pre-POINT randomized clinical trial. JAMA. 2015;313:1541-1549.
18. 18. Roep B. Primary prevention for type 1 diabetes mellitus? Nat. Rev. Endocrinology 2015;11:451-452.
19. 19. Zhang ZJ, Davidson L, Eisenbarth G, Weiner HL Zhang J. Suppression of diabetes in nonobese diabetic mice by oral administration of porcine insulin. Proc Natl Acad Sci USA 1991;88:10252-10256.
20. 20. Kamyshny A, Putilin D, Kamyshna V. Reduced deaf1 mRNA expression during STZ-induced diabetes mellitus inhibits foxp3+regulatory T-cells differentiations in rat’s pancreatic lymph nodes. Mediterranean J of Biosciences 2015; (1):20-26.