Effect Of Tim-1 And Tim-3 On Th1/Th2 And Th17/Treg Cells Imbalances In Children With Henoch-schonlein Purpuras

objective: The aim of this study was to investigate the changes of the levels of Tim-1?Tim-3 in children with HSP in acute phase and the possible roles of Tim-1 and Tim-3 in pathogenesis in acute phase of HSP,to provide experimental basis for further understanding the pathogenesis of HSP from Tim-1,Tim-3 perspective;to investigate the immunomodulatory effect of Tim-1 and Tim-3 in Th1 / Th2 cells and Th17 / Treg cells imbalances in acute phase of HSP,and to provided the experimental basis to further illustrate the molecular mechanism of Th1/Th2 cells and Th17/Treg cells imbalances.Methods:A total of 45 children with HSP in acute phase admitted in Sichuan Provincial People's Hospital from July 2014 to May 2015 were enrolled in the experimental group,and 45 healthy children were enrolled in the control group.Mononuclear cells(PBMC)were isolated from peripheral blood by density gradient centrifugation,total RNA was extracted from PBMC by Trizol method,and then reverse transcribed into cDNA.The expression levels of target genes Tim-1 mRNA,Tim-3 mRNA and ROR?T mRNA were analyzed by SYBR?Premix Ex Taq TM II fluorescence quantitative PCR.The levels of serum cytokines IFN-?,IL-4,IL-10 and IL-17 A were determined by ELISA.The expression of CD4 + CD25 + Treg cells in T cell subsets was measured by flow cytometry.Results: The results showed that in HSP group the expression levels of Tim-1 mRNA(1.33�0.261),Tim-3 mRNA(1.53�0.247)and ROR?t mRNA(0.81�0.097)were significantly higher than those of controlgroup [Tim-1 mRNA(0.84�0.096),Tim-3 mRNA(0.73�0.091)and ROR?t mRNA(0.46�0.199)respectively p<0.01)].In HSP group the levels of serum IL-4,IL-17A(44.52�16.66 pg/ml,58.32�14.53 pg/ml respectively)were higher than those in the control group(18.32�5.43 pg/ml,31.79�12.80 pg/ml respectively)(P<0.01),while the levels of serum IFN-?,IL-10(32.96�15.14pg/ml,14.60�6.44 pg/ml respectively)were lower than those of control group(49.82�20.60 pg/ml,21.78�7.42 pg/ml respectively)(P<0.01).Compared with the control group(10.06�1.63%),the level of CD4+CD25+ Treg cells in HSP group(6.95�2.33 %)was decreased(P<0.01).In the HSP group,the relative expression level of the target gene Tim-1 mRNA was positively correlated with IL-4,the correlation coefficient was 0.398(P <0.01);and was negatively correlated with IFN-? and CD4+CD25+Treg cell(P <0.01),the correlation coefficients were-0.453(P<0.01)and-0.365(P<0.05)respectively,while there was no correlation between Tim-1 mRNA with IL-17 A and ROR ?t mRNA,the correlation coefficients were 0.229(p = 0.13> 0.05)and 0.087(P = 0.571> 0.05)respectively.The relative expression level of the target gene Tim-3 mRNA was positively correlated with the expression levels of CD4+CD25+Treg cell and IL-4,the correlation coefficients were 0.404(P<0.01)and 0.626(P<0.01)respectively,while was negatively correlated with levels of IFN-??IL-17 A and ROR ?t mRNA,the correlation coefficients were-0.611(P<0.01)?-0.478(P<0.01)?-0.321(P<0.05)respectively.Conclusion:1)the expression level of IFN-? was down-regulated and the expression level of IL-4 was up-regulated,suggesting that there were Th1 / Th2 cell imbalance in the acutephase of HSP,and Th2 cells predominated over Th1 cells;the expression levels of IL-17 A and ROR ?t mRNA were increased and the expression levels of IL-10 and CD4+CD25+Treg cell were decreased,suggesting that there were Th17/Treg cell imbalance in the acute phase of HSP,and Th17 cells predominated over CD4+CD25+Treg cells.2)The expression leves of Tim-1mRNA and Tim-3 mRNA in PBMC of HSP group increased,suggesting that Tim-1 and Tim-3 may play a role in the pathogenesis of HSP in acute phase.3)The relative expression level of Tim-1 mRNA was positively correlated with the level of IL-4 and negatively correlated with the levels of IFN-? and CD4 +CD25 + Treg cells,the relative expression level of Tim-3 mRNA was positively correlated with levels of IL-4 and CD4 + CD25 + Treg cell,while was negatively correlated with the levels of IFN-?,IL-17 A and ROR ?t mRNA.It showed that the expression changes of Tim-1 and Tim-3 may taken a place in the pathogenesis of the Th1/Th2 cells imbalance and Th17/Treg cells imbalance of HSP in acute phase.