Regulation Of SOCS1/3 To TLR3 And Tfh Cell Differentiation And Its Role In HT

Objective:1.To investigate the roles and the specific mechanisms of SOCS1,SOCS3 and TLR3 in HT diseases,we tested the expression of SOCS1,SOCS3 and TLR3 protein in thyroid tissue and m RNA in peripheral blood from patients with HT.2.To investigate the role of CD4+CXCR5+T(Tfh)cell in the onset of HT,we detected the percentage of Tfh cells in peripheral blood lymphocytes from HT patients,and tested the expression of its characteristic transcription factors Bcl6 m RNA in peripheral blood.Method:1.The expression and location of SOCS1/3 and TLR3 protein in thyroid tissues which selected form 30 cases of HT,10 cases of GD and 10 cases of normal were detected by immunohistochemistry staining(IHC),and the results were analyzed by quantitative analysis method.2.We collected the peripheral bloods from 30 cases of HT,30 cases of GD and 30 cases of normal,respectively.Following the mononuclear cells(PBMC)were isolated from peripheral blood by using Ficoll density gradient centrifugation,and the total RNA was extracted from lymphocytes.Then the relative expression quantity of SOCS1/3 and TLR3 m RNA in peripheral blood PBMC from three groups of patients were examined by applying real-time PCR.3.The percentage of Tfh cell in fresh peripheral blood lymphocytes from 30 cases of HT,30 cases of GD and 30 cases of normal were detected by using flow cytometry.The mononuclear cells(PBMC)were isolated from peripheral blood by using Ficoll density gradient centrifugation,and the relative expression quantities of Bcl6 m RNA were examined by applying real-time PCR.Results:1.The TLR3 protein was hardly expressed in normal and Grave's thyroid tissues,but in HT thyroid tissues it is mainly expressed in cytoplasm of follicular epithelial cells.The expression of TLR3 protein in HT group is obviously higher than that in normal and GD group,the difference was statistically significant(P < 0.05).2.There is a little expression of SOCS1/3 protein in normal and Grave's thyroid tissues,and it mainly expressed in cytoplasm of follicular epithelial cells.The expression quality of SOCS1/3 protein in HT group is higher than that in normal and GD group,the difference was statistically significant(P < 0.05).3.The relative expression quantity of TLR3,SOCS1/3 m RNA in HT group is higher than normal and GD group,the difference was statistically significant(P < 0.05).4.The percentage of Tfh cells in peripheral blood of HT group is higher than that of GD group and normal group,the difference was statistically significant(P < 0.05).The relative expression quantity of the characteristic transcription factors Bcl6 m RNA for Tfh cells in peripheral blood PBMC from HT group is higher than the other two groups.The difference was statistically significant(P < 0.05).5.The relative expression quantity of SOCS1 m RNA in PBMC of peripheral blood from HT patients is negatively associated with the expression level of TGAb in serum(r=0.393),while there is no relationship between the expression of SOCS1 m RNA and TPOAb,FT3,FT4,TSH(P > 0.05).6.The relative expression quantity of TLR3,SOCS3 m RNA in PBMC of peripheral blood from HT patients is not associated with TGAb,TPOAb,FT3,FT4 and TSH in serum(P > 0.05).7.The differentiation percentage of Tfh cells in peripheral blood of HT patient is positively associated with TGAb(r=0.380)and TPOAb(r=0.374)in serum,while has no correlation with FT3,FT4 and TSH in serum(P>0.05).Additionally,the relative expression quantity of Bcl6 m RNA in HT patient peripheral blood is not associated with TGAb and TPOAb(P > 0.05).8.There is no correlation between the expressions of TLR3 protein and SOCS1/3 protein in thyroid tissues from HT patients(P > 0.05).9.The relative expression quantity of SOCS1 m RNA is positively associated with the expression quantity of TLR3 m RNA in PBMC of peripheral blood from HT patients(r=0.586,P=0.001).Meanwhile,the expression of SOCS3 m RNA is not associated with the expression of TLR3 m RNA in PBMC of peripheral blood from HT patients(P > 0.05).10.The relative expression quantity of SOCS1/3 m RNA is not associated with the percentage of Tfh cells in HT group(P > 0.05).Also there is no correlation between the relative expression quantity of SOCS1/3 m RNA and Bcl6 m RNA(P > 0.05).Conclusion:1.The expressions of TLR3,SOCS1 and SOCS3 in thyroid tissue and peripheral blood from patients with HT are higher than normal and GD group,which indicating that SOCS1,SOCS3 and TLR3 may be associated with the onset of HT.2.The percentage of Tfh cells in peripheral blood and the relative expression quantity of the characteristic transcription factors Bcl6 m RNA for Tfh cells in peripheral blood PBMC from HT group are higher than that of GD group and normal group.This indicates that Tfh cells may participate in the onset of HT.3.The percentage of Tfh cells in peripheral blood from HT patients and autoantibodies TGAb,TPOAb in serum were positively correlated,prompting that Tfh cells may be associated with B cells to producing antibodies of HT.