Introduction: Mesenteric lymph nodes (MLN) is a major transition point for recirculating lymphocytes of GALT. Homing of lymphocytes in MLN is regulated with adressin MAdCAM-1, chemokine receptors CXCR4 and CCR7. Sphingosine-1-phosphate receptors S1PR1 activate T-cell exit from MLN.
Methods: We use RT-PCR method for investigating of mRNA expression levels of genes MAdCAM-1, CXCR4, CCR7 and S1PR in MLN of the offspring of rats with experimental gestational diabetes. To determine the level of target genes was performed RT-PCR in real-time by thermocycler CFX96 Real-Time PCR Detection Systems. The relative level of gene expression were studied with rat reference genes GAPDH by the method ΔΔCt. Statistical analysis were conducted using available software «Bio-Rad CFX Manager 3.1» (Bio-Rad, USA).
Results: Expression analysis of homing receptors in MLN revealed an expected significant increasing of CCR7 and MAdCAM-1 mRNA in offspring of animals with EGD, indicating the activation of the immune cells in the GALT, which is accompanied by intensification of lymphocytes homing and confirms the involvement of these receptors in the pathogenesis of diabetes. We were unable to detect changes in the mRNA levels of another regulator CXCR4 in MLN of the offspring of rats with EGD. Increased expression level of S1PR1 mRNA of MLN lymphocytes in the offspring of animals with diabetes confirms its important role in the progression of diabetes.
Conclusions: Signals of chemokine receptors affect the activation of different Th cells subsets and we may assume their pivotal role in the development of autoimmune diseases, particularly diabetes mellitus. The revealed changes evidence of abuse of formation of peripheral immunological tolerance and can trigger the development of AID in the offspring of mothers with EHD.
20 - 23 May 2017
European Society of Endocrinology