Relativity Analysis Of Th17/Treg Balance And IL-35 In Oral Lichen Planus Patients

Objective: To investigate the variations of Th17/Treg immune balance and the expression of IL-35 in oral lichen planus(OLP) patients peripheral blood and focal mucosa,and to explore the relationship between IL-35 and Th17/Treg balance and these possible roles in the pathogenesis of OLP. Methods: 1. Forty OLP patients(23 patients with non-erosive OLP and 17 patients with erosive OLP) and 15 healthy controls were enrolled in the study. The levels of serum IL-17 and TGF-β1 were measured using enzyme-linked immunosorbent assay. The m RNA expression of RORc, FOXP3, IL-17, Ebi3, and p35 in peripheral blood mononuclear cells(PMBCs) were detected by real-time fluorescent quantitative PCR(q PCR). The correlations between the above molecule expressions and clinical characteristics of patients with OLP were analyzed, respectively. 2. Forty-one cases of OLP(20 cases of non-erosive OLP and 21 cases of erosive OLP) and 12 normal oral mucosa specimens were included. The expressions and distributions of proteins about IL-17, FOXP3, TGF-β1 and Ebi3 in OLP lesions were examined by immunohistochemical staining(IHC). The m RNA expression levels of RORc, FOXP3, IL-17 and IL-35 subunits(p35/EBI3) in OLP lesions were evaluated through quantitative real-time PCR. The correlations between the above molecule expressions and clinical characteristics of patients with OLP were analyzed, respectively. Results: 1. The levels of serum IL-17 were significantly higher in erosive OLP patients compared to healthy controls and non-erosive OLP patients(P<0.05) and a positive correlations of IL-17 level with the scoring of signs were observed(r=0.311, P=0.017).But no statistical differences were observed in the serum levels of TGF-β1 between OLP patients and normal controls(P>0.05). Meanwhile, q PCR result showed that the m RNA expression of IL-17,RORc, FOXP3 and IL-35 subunits p35 and EBI3 were prominently elevated in OLP patients compared with those in the control group respectively(P<0.05). Considering the subtypes of the OLP groups, m RNA expression of FOXP3 was dramatically higher in non-erosive OLP patients than in erosive OLP patients, while others showed no statistical differences of these arguments between the reticular form group and the erosive form group. Additionally, RORc/FOXP3 ratio in erosive OLP was significant higher than that in control group, moreover, increased RORc/FOXP3 ratio in non-erosive OLP was found compared to control group, whereas there was no marked difference were indicated. The Spearman correlation analysis showed that both FOXP3 and p35 expressions were negatively correlated with the RAE score and the scoring of signs and the m RNA expressions of IL-17, p35 and EBI3 were positively correlated with RORc and FOXP3 respectively in PBMCs of OLP patients. 2. IHC statistical analysis showed that the expression levels of IL-17, FOXP3, Ebi3 and TGF-β1 were sharply increased in OLP groups compared with that in controls, and protein Ebi3 in lesions with OLP was related to liquefaction degeneration of basal cells. Meanwhile,The m RNA levels for Th17 and Treg molecules(IL-17, RORc and FOXP3, Ebi3 and p35) in OLP lesions were higher than those in controls(P<0.05). The RORc/FOXP3 ratio was increased in erosive OLP lesions when compared with controls, nevertheless, there was no significant correlations were observed between the non-erosive OLP and control groups. Conclusion:(1) Th17 and Treg cell levels were elevated in peripheral blood and lesions tissues in OLP patients and there was Th17/Treg imbalance in the erosive OLP, but not in non-erosive OLP.(2) The upregulation of IL-35 has an immune regulatory effect on Th17/Treg balance, which is related to the formation of chronic inflammation in OLP.