Effects Of IL-35 On Th17/Treg Balance In Peripheral Blood Of Patients With Oral Lichen Planus

Objective: To study the condition of Th17/Treg balance in peripheral blood of patients with oral lichen planus(OLP)and to investigate the effects of IL-35 on Th17/Treg balance in peripheral blood of patients with OLP.Methods: 1.12 cases of patients with OLP(4 cases of reticular OLP and 8 cases of erosive OLP)were enrolled in this study,and 13 cases of healthy people served as controls.Peripheral blood mononuclear cells(PBMCs)were extracted in sterile and CD4+T cells were sorted by flow cytometry(FCM),the expression of Foxp3 and ROR?t m RNA were detected by real-time PCR.The cells were cultured in vitro by adding / without adding rIL-35.After culture finished,the cells were collected and the same factors were tested by real-time PCR.2.15 cases of patients with OLP(6 cases of reticular OLP and 9 cases of erosive OLP)were enrolled in this study,and 14 cases of healthy people served as controls.Peripheral blood mononuclear cells(PBMCs)were extracted in sterile and CD4+CD25+T cells were sorted by flow cytometry(FCM),the expression of Foxp3,EBI3,p35 mRNA and IL-10 were detected by real-time PCR and ELISA,respectively.The cells were cultured in vitro by adding / without adding rIL-35.After culture finished,the cells and cells culture supernatant were collected and the same factors were tested by real-time PCR and ELISA.Results: 1.(1)The expression of Foxp3 and ROR?t mRNA in CD4+T cells of patients with OLP were higher than normal controls(p<0.05),the ratio of ROR?t/Foxp3 mRNA in erosive OLP group was higher than that in reticular OLP group and control groups(p<0.05).Spearman correlation analysis showed there was a positive correlation between the ratio of ROR?t/Foxp3 and symptom score(r=0.661,p=0.019).(2)The expression of Foxp3 mRNA was down-regulated after adding rIL-35 in normal groups(p<0.05),the expression of ROR?t mRNA showed no significant difference(p>0.05),and the ratio of ROR?t/Foxp3 was increased after adding rIL-35(p<0.05).(3)The expression of Foxp3 mRNA was up-regulated after adding rIL-35 in CD4+T cells of erosive OLP group(p<0.05),the expression of ROR?t mRNA showed no significant difference(p>0.05),and the ratio of ROR?t/Foxp3 was declined after adding rIL-35(p<0.05).2.(1)The expression of Foxp3,EBI3,p35 mRNA and IL-10 in peripheral blood Tregs cells of patients with OLP were higher than those in normal controls(p<0.05).(2)The expression of Foxp3 mRNA and IL-10 were down-regulated after adding rIL-35 in normal groups(p<0.05),the two subunits EBI3 and p35 of IL-35 also showed a decline in mRNA levels(p<0.05).(3)The expression of Foxp3 m RNA and IL-10 were up-regulated after adding rIL-35 in peripheral blood Tregs cells of patients with OLP(p<0.05),the two subunits EBI3 and p35 of IL-35 also showed an increase in mRNA levels(p<0.05).Conclusion: 1.The expression of Th17 and Tregs cells in the peripheral blood of OLP was increased,and it was involved in the pathogenesis of OLP,and there was a Th17/Treg imbalance with predominanted by Th17 cells,which was associated with long-term chronic inflammation of OLP.2.Exogenous IL-35 has a different effect on both healthy groups and OLP patients.In healthy controls,IL-35 can downregulate the expression of Tregs cells and secretion of IL-10 and IL-35,while the expression of Tregs cells and their secretion of IL-10 and IL-35 were up-regulated in OLP,suggesting that exogenous IL-35 has an immunoregulatory effect on Th17 / Treg balance,and it is speculated that this difference may be one of the causes of OLP.