Preliminary Study On The Function Of Peripheral Blood CD4~+NKG2D~+IL17~+T Cell In NOD Mice

Type ? diabetes is a self-reactive bovine cell-mediated autoimmune disease,the main mechanism for autoimmune lymphocyte infiltration of islets to kill islet ? cells,resulting in absolute lack of insulin secretion leading to diabetes.Type ? diabetes is dominated by CD4+T cells and CD8+ T cells,with a variety of lymphocytes such as NK cells,NKT cells,??T and other cells involved in the process of type I diabetes.In recent years,studies at home and abroad have confirmed that NKG2D expression is not only expressed on the surface of NK cells in NOD mice,but also on the surface of CD4+ T cells and CD8-positive T cells,while Th17 cells play an important inflammatory condition in NOD mice with type I diabetes effect.The expression of NKG2D and IL-17 in CD4 + T cells of NOD mice and the expression of other cytokines and nuclear transcription factors were detected by flow cytometry.The function and phenotype of CD4 +NKG2D + T cells were deduced.Part 1:Expression of CD4 + T cells in NOD mice NKG2D is associated with the duration of type 1 diabetes mellitusObjective:To monitor the changes of blood glucose and the frequency of CD4+ NKG2D+T cells in NOD mice,and to elucidate the relationship between CD4+ NKG2D+ T cells and type I diabetes mellitus.Methods:Peripheral blood glucose changes were monitored weekly at 12 weeks of age in NOD mice.The frequency of CD4 + NKG2D + T cells in peripheral blood of mice was detected by flow cytometry at 8w,10w,12w and 14w every 2 weeks.The frequency of CD4 + NKG2D+/CD4 + cells was counted and the correlation between NKG2D-expressing CD4 + T cells and type? diabetes mellitus was analyzed.Results:The blood glucose of NOD mice increased gradually with the disease and reached the highest at 28 weeks of age.The morbidity statistics showed that the onset of the mice at 12 weeks of age was the highest at 28 weeks of age.Flow cytometry analysis CD4 + NKG2D + T cells with the development of type I diabetes gradually increased in the incidence of cell frequency reached the highest.Indicating that CD4+ NKG2D+ T cells are involved in the progression of NOD disease.Part ?:Characterization and functional analysis of NKG2D+CD4+T cellsObjective:To cdetect tthe expression and secretion of cytokines in CD4 + NKG2D + T cell membrane and to speculate the function and phenotype of CD4 + NKG2D + T cells.Methods:The expression of CD4 + NKG2D + T cell membrane and the expression of IL-17,ESNF?,IL10 and other cytokines were detected by flow cytometry at 15w,20w,25w and 30w weeks of age.Results:The level of IL17 secretion in the progression of NOD mice was gradually increased to the highest level.IFN? disease prodromal period and the level of secretion has been in a high state.FasL,CD 107a in the disease progression and disease stage low secretion or no secretion,suggesting that CD4 + NKG2D + T cells without cytotoxic activity.IL10 secretion levels in the entire course of the process of low secretion may not have regulatory activity.CD25 and CD69 surface protein were expressed,indicating that CD4 + NKG2D + T cells in the activation state.The level of IL-1 secreted by CD4 + NKG2D + T cells in the prophylaxis and disease period was similar,which was presumably similar to that of Th17 cells.CD4 + NKG2D+ T cells do not have cytotoxic and immunomodulatory effects.Part ?:Characteristic analysis of CD4+ NKG2D+IL17+ T cells Objective:To investigate the relationship between CD4+ NKG2D+ T cells and Th17 cells by analyzing the characteristics of CD4 + NKG2D+IL17+ T cells by flow cytometry.Methods:The transcription factors ROR?T,intracellular factor TGFO,and membrane molecules NK1.1,CCR6 were detected by flow cytometry in peripheral blood of mice CD4 +NKG2D+T cells according to the pathogenesis of mouse disease after 30 weeks And to predict the relationship between CD4+ NKG2D+IL17+ T cells and Th 17 cells.Results:The expression of RORyT in peripheral blood CD4 + NKG2D + T cells of NOD mice increased gradually,reached the peak at the onset stage,and the level of TGF? was lower,which was not statistically significant due to individual differences.Disease prophase phase and disease phase compared to CCR6 secretion levels increased.The expression of NK1.1 in CD4 +NKG2D + T cells was significantly higher than that in CD4 + NKG2D + NK1.1-cells.Thus,CD4+NKG2D+ T cells are a group of cells that function similarly to Thl7 cells.The expression of RORyT in prophylaxis and disease stage,but also promote the low secretion of TGF? in Th17 cells.CD4 + NKG2D+ NK1.1 is mainly negative and high expression of CCR6.It can be deduced that CD4+ NKG2D+ T cells play an important role in the pathogenesis of type 1 diabetes mellitus Role,and similar to the function of Th17 cells,but not the same group of cells.