To Investigate The Effect Of TRAF-6 On IL-2 Induced Immune Tolerance In Graves' Disease

PART1 THE ROLE OF IL-2 IN THE REGULATION OF TH17/TREG CELL BALANCE IN THE PATHOGENESIS OF GRAVES' DISEASEObjective: To investigate the role of IL-2 in the regulation of Th17/Treg cell balance in the pathogenesis of Graves' disease.Methods: Thirty GD newly diagnosed patients(GD),30 GD patients in remission(eGD)and 28 normal controls(NC)were enrolled in this study.Thyroid function(FT3,FT4,TSH)and autoantibodies(TRAb,TPOAb,TgAb)were detected by chemical immunofluorescence.Peripheral blood mononuclear cells(PBMCs)were extracted and real-time PCR was used to detect the expression of ROR?t,Foxp3 and IL-2 mRNA.The plasma expression of IL-2,IL-17 and IL-10 protein were determined by ELISA.Results: 1.The thyroid function of GD group was significantly higher than that of eGD and NC group(P<0.05).The levels of TRAb and TgAb were significantly higher in GD group than in eGD and NC group,and the levels of TRAb and TgAb were significantly higher in eGD group than in NC group(P<0.05).Compared with the NC group,the levels of TPOAb in GD group and eGD group were significantly higher(P<0.05),but there was no significant difference in TPOAb level between GD group and eGD group(P>0.05).2.Compared with the NC group,the expression of IL-2 mRNA and protein,Foxp3 mRNA and IL-10 protein in the GD group decreased,and the expression of ROR?t mRNA and IL-17 protein increased,and the above results are statistical differences(P<0.05).3.In the GD group,the expression level of IL-2 mRNA and IL-17 protein was positively correlated(P<0.05),and the expression level of IL-2 mRNA was not correlated with the expression of ROR?t mRNA,Foxp3 mRNA and IL-10 protein,respectively(P>0.05),and the expression level of IL-2 protein was not correlated with the expression of ROR?t mRNA,IL-17 protein,Foxp3 mRNA and IL-10 protein,respectively(P>0.05).Conclusion: The decreased expression of IL-2 leads to the increased differentiation of Th17 cells and the decreased differentiation of Treg cells,which cannot effectively regulate the balance of Th17/Treg cells and promote the occurrence of GD.PART2 THE IMMUNE TOLERANCE OF TRAF-6 IN GRAVES' DISEASE AND ITS POTENTIAL MECHANISMObjective: To investigate the immunotolerant role of TRAF-6 in Graves' disease and its potential mechanism.Methods: Thirty GD newly diagnosed patients(GD),30 GD patients in remission(e GD)and 28 normal controls(NC)were enrolled in this study.PBMC were extracted and the expression of TRAF6 m RNA was detected by real-time PCR.The protein expression of p-Smad2,p-Smad3 and TRAF6 were detected by Western blot.ELISA was used to detect the expression of TGF-?1 protein in plasma.Results: 1.Compared with the NC group,the expression of TGF-?1 protein in the GD group was decreased,and the difference was statistically significant(P<0.05).The expression of p-Smad2 and p-Smad3 protein in the GD group were higher than those in the NC group.(P>0.05).2.The expression of TRAF-6 m RNA in GD group was lower than that in NC group(P<0.05).The expression of TRAF-6 protein was lower than that in NC group(P>0.05).3.In the GD group,there was no correlation between the expression of TRAF-6 m RNA and p-Smad2 protein,p-Smad3 protein respectively(P>0.05);And the expression level of TRAF-6 protein and p-Smad2 protein were positively correlated(P<0.05),and the expression level of TRAF-6 protein was not correlated with p-Smad3 protein(P>0.05);In the GD group,the expression of p-Smad2 protein and IL-2 m RNA were negatively correlated(P<0.05),and there was no correlation between the expression of p-Smad2 protein and IL-2 protein(P>0.05);There was no correlation between the expression of p-Smad3 protein and the expression of IL-2 m RNA,IL-2 protein respectively in the GD group(P>0.05).Conclusion: TRAF-6 plays an immunotolerant role in the development of GD;the low expression of TRAF-6 is not sufficient to inhibit the phosphorylation of Smad2/3 by TGF-?1,resulting in increased phosphorylation of Smad2/3,down-regulating the expression of IL-2,inducing the differentiation of Th17 cells and inhibiting the differentiation of Treg cells,which makes the body unable to maintain immune tolerance and promote the development of GD.