Detection Of Regulatory T Cell And Its Functional Phenotype In Peripheral Blood Of Patients With Autoimmune Thyroid Disease And Its Clinical Significance

Objective1.To observe the proportion of Treg(CD3 + CD4 + CD25 + Foxp3 +)cells in peripheral blood of patients with autoimmune thyroid disease,and to explore the correlation between the etiology of autoimmune thyroid disease and the number of Treg cells.2.To observe the expression of Helios +,PD-1 +,aTreg(CD3 + CD4 + CD25 + Foxp3hiCD45RO+).sTreg(C D 3 + C D 4 + C D 2 5 + F o x p 3 1o C D 4 5 R O +)?r T r e g(CD3+CD4+CD25+Foxp310CD45RO-)in peripheral blood Treg cells of patients with autoimmune thyroid disease.To investigate the relationship between autoimmune thyroid disease and Treg cell function.3.To observe the distribution of Helios + and PD-1 + cells in peripheral blood aTreg,sTreg and rTreg cells of patients with autoimmune thyroid disease,and to explore the correlation between autoimmune thyroid disease and Treg cell function in different subtype.4.To observe the ratio of CD45RO + T effect and CD45RO-T effect in peripheral blood T effect cells of patients with autoimmune thyroid disease,and the distribution of PD-1 +,Helios + cells in CD45RO + T effect and CD45RO-T effect,to explore the changes of effector T cells during the pathogenesis of autoimmune thyroid disease.Method:Take the normal control,Graves' patients,Hashimoto thyroiditis patients with peripheral blood 3-4ml,Ficoll density gradient centrifugation was used to separate PBMCs.The cells were stained with flow cytometry(FCM)and stained on the cell surface and intracellularly.Step by step detection:(1)The percentage of Foxp3 +CD25 + Treg?Teffect in peripheral blood CD4 + T cells(2)Percentage of PD-1 +,Helios + cells,aTreg,sTreg,rTreg in Foxp3 + CD25 + Treg cells;(3)Detection of Helios +,PD-1 + in aTreg,sTreg and rTreg cells,respectively(4)Detection of memory Teffect cells and initial Teffect cells in Teffect cells,the percentage of PD-1+ Helios + cells in the memory Teffect cells and initial Teffect cells was measured respectively.Result:1.CD3 + CD4 + Foxp3 + T cells accounted for CD3 + CD4 + T cells in the Hashimoto thyroiditis group was higher than the control group,it's(7.34 ± 2.35)%vs(5.53 ± 1.43)%,respectively(P<0.05),the difference was statistically significant;Graves disease patients and control group and Hashimoto thyroiditis group no significant difference(P>0.05).2.The expression of CD3 + CD4 + CD25 + Foxp3 + Treg cells in CD3 + CD4 + T cells was significantly higher than that in control group(5.23 ± 1.57)%vs(4.25 ±1.38)%,(P<0.05)in patients with Hashimoto thyroiditis,the difference was statistically significant;Graves disease patients and control group and Hashimoto thyroiditis group no significant difference(P>0.05).3.The average fluorescence intensity of the transcription factor Foxp3 in patients with Hashimoto thyroiditis was higher than that in the control group 1213.88 ±384.41 vs 911.85 ± 229.33(P<0.05),the difference was statistically significant.There was no significant difference between Graves disease and control group and Hashimoto thyroiditis group(P>0.05).4.The proportion of PD-1 + cells in Treg cells was higher in patients with Hashimoto thyroiditis than in the control and Graves' patients,it's(30.90 ± 9.74)%vs(18.93 ± 7.38)%(P<0.05),(30.90 ± 9.74)%vs(23.75 ± 10.06)%(P<0.05).The difference was statistically significant.Graves disease patients and the control group no significant difference(P>0.05).5.The percentage of aTreg cells in Treg cells was significantly higher in Hashimoto thyroiditis than that in control group(16.38 ± 6.44)%vs(11.68 ± 6.97)%(P<0.05),and the difference was statistically significant.There was no significant difference between Graves disease and control group and Hashimoto thyroiditis group(P>0.05).6.The proportion of PD-1 + cells in sTreg cells was higher in patients with Hashimoto thyroiditis and Graves disease than in the control group,The expression levels were(33.76 ± 6.22)%vs(23.52 ± 8.93)%(P<0.05),(32.51 ±8.88)4%vs(23.52 ± 8.93)%,respectively(P<0.05).The difference was statistically significant;Graves disease patients with Hashimoto thyroiditis group no significant difference(P>0.05).7.The expression of sTreg cells in the total number of CD4 + T cells in Hashimoto thyroiditis was higher than that in the control group(16.38 ± 6.44)%vs(11.68 ±6.97)%(P<0.05),and the difference was statistically significant.There was no significant difference between Graves disease and control group and Hashimoto thyroiditis group(P>0.05).8.The expression of aTreg cells in the total number of CD4 + T cells in Hashimoto thyroiditis was higher than that in the control group(0.84±0.36)%vs(0.50±0.37)%(P<0.05),and the difference was statistically significant.There was no significant difference between Graves' patients and control group and Hashimoto thyroiditis group(P>0.05).9.CD45RO + Teffect cells accounted for T effect cells in the Hashimoto thyroiditis group was higher than the control group(52.91 ± 11.75)%vs(35.89 ±11.13)%(P<0.05),the difference was statistically significant;there was no significant difference between Graves' patients and control group and Hashimoto thyroiditis group(P>0.05).10.CD45RO-Teffect cells accounted for T effect cells in the Hashimoto thyroiditis group was lower than the control group(47.09±11.78)%vs(64.13±11.15)%(P<0.05),the difference was statistically significant;there was no significant difference between Graves'patients and control group and Hashimoto thyroiditis group(P>0.05).11.The percentage of PD-1 + cells in CD45RO + Teffect cells in the Hashimoto thyroiditis group was higher than that in the control group(47.46 ± 9.28)%vs(37.68 ± 9.72)%(P<0.05),the difference was statistically significant.There was no significant difference between the Graves' and the control group and Hashimoto thyroiditis group(P>0.05).Conclusion:The proportion of peripheral blood Treg cells in Hashimoto thyroiditis patients was higher than that in the normal control group,especially the aTreg,but there was no significant change in the patients with Graves disease,which indicated that it played a role in Hashimoto thyroiditis immunological damage.The expression of PD-1 was also increased in the peripheral blood of patients with Hashimoto thyroiditis.However,the expression of PD-1 on the surface of memory-induced T cells was increased in patients with Graves disease.Suggesting that the number and functioa of Treg cells were mainly performed in patients with Hashimoto thyroiditis,The early onset of Graves mainly for Thl/Th2 cell imbalance.