A Preliminary Study On The Interaction Of CXCL10-CXCR3/CCL17-CCR4 In The Pathogenesis Of Oral Lichen Planus

Background: At present,the specific pathogenesis of oral lichen planus(OLP)is still unclear.The pathogenesis of OLP may include immune,mental,endocrine,infectious and genetic factors.Previous studies of this research group and related literature reports mainly focused on the exploration of the expression of single chemokine-receptor axis in peripheral blood and tissues of OLP patients,but there are few studies on the correlation between the chemokine-receptor axis.Literature reports have pointed out that chemokines and their receptors play a certain role in the pathogenesis of OLP,but there is no relevant report on the cross-linking effect of each chemokine-receptor axis in the complex chemokine immune regulation network of OLP.Objective: Through antagonistic intervention of peripheral blood T cells,flow cytometry and enzyme-linked immunosorbent assays to detect the expression of related receptors and ligands,and in vitro culture of human T lymphocyte lines for stimulation and reverse verification,to preliminarily explore the onset of OLP What is the correlation and interaction between the CXCL10-CXCR3 and CCL17-CCR4.Methods:1.Pass11 software and literature review were used to determine the clinical sample size,and peripheral blood was collected from OLP patients(non-erosive,erosive group)and healthy controls,and T lymphocytes were separated and identified for purity.T cells were co-cultured with three groups: blank,anti-CXCR3 and anti-CCR4.2.Flow cytometry was used to examine the expression of CXCR3 and CCR4 on the surface of T lymphocytes in OLP patients(non-erosive and erosive group)and healthy controls.The correlation between chemokine receptor CXCR3 and CCR4 was preliminarily investigated.3.Enzyme-linked immunosorbent assay(ELISA)was used to monitor the level of the corresponding ligands CXCL10 and CCL17 in each group.The changes in ligand trends were verified and the correlation between the two chemokine receptor axes was speculated.4.In vitro experiments on human H9 T lymphocyte lines were used to verify the above results,and CCK8 method was used to determine the stimulation concentration and time of chemokines CXCL10 and CCL17 in each group.Flow flow and ELISA were used to detect the results.Graph Pad 6 software was used to analyze and compare the expression results of OLP patients and healthy controls,and the data of each group were statistically processed and analyzed.Results:1.The purities of CD3+T cells were all >95% in the peripheral blood of OLP patients(non-erosive,erosive group)and controls(P>0.05).And there was no statistically prominent difference between the groups.2.Flow cytometry was used to detect the expression of CXCR3 and CCR4 receptors.Compared with the blank group,the expression of anti-CXCR3 and anti-CCR4 group in OLP was down-regulated(P>0.05).The expression of CCR4 in antagonistic CCR4 group was significantly down-regulated(P<0.05),and the expression of CXCR3 was up-regulated(P>0.05).The receptor and ligand trends of healthy controls were consistent with those of OLP,but there was no remarkable difference between the anti-CXCR3 or anti-CCR4 group and the blank group(P>0.05).The consequences of CXCR3 and CCR4 in OLP patients and healthy controls were compared,and there were statistically significant differences in the blank group,antagonistic CXCR3 and CCR4 groups(P<0.05).3.Enzyme linked immunosorbent assay(ELISA)was used to detect the expression of ligand CXCL10 and CCL17.Compared with blank group,the level of CXCL10 in anti-CXCR3 or anti-CCR4 group was notably down-regulated in OLP(P<0.05),the situation of CCL17 was also down-regulated(P>0.05).The expression of CCL17 in anti-CCR4 group was dramatically down-regulated(P<0.05),and the level of CXCL10 was up-regulated(P>0.05).The receptor and ligand trends of healthy controls were consistent with those of OLP,but there was no significant difference between the antagonist group and the blank group(P>0.05).Comparing the CXCL10 results of OLP patients with healthy controls,there was statistically significant difference between the blank group and the anti-CCR4 group(P<0.05),while there was no statistically striking difference between the anti-CXCR3 group(P>0.05).The level of CCL17 showed that the difference in blank group was statistically noteworthy(P<0.05),while there was no statistically significant difference in antagonistic CXCR3 and CCR4 groups.4.The results showed that the purity of human H9 CD4+T lymphocyte lines were all >95%.CCK8 method confirmed that the optimal concentration and time of CXCL10 and CCL17 were 50ng/ml,96 h and 100ng/ml,72 h,respectively.Flow cytometry and ELISA conclusions indicateded that,the levels of CXCL10 and CXCR3 were increased in the CXCL10 stimulation group,and the expression of CCR4 was also increased,compared with the blank group.While CCL17 had no significant change.After CCL17 stimulation,CXCR3 expression decreased,CCR4 expression increased,CXCL10 and CCL17 expression did not change significantly.After statistical analysis,there was no statistical significance among all groups.Conclusions:1.There are related interactions between chemokine receptors CXCR3 and CCR4 in the peripheral blood of OLP patients,suggesting mixed inflammatory responses in the local immune microenvironment of OLP.2.The chemokine ligands CXCL10,CCL17 and the corresponding receptors interact with each other in the chemokine immune regulation network,and each plays a certain role.3.CXCL10-CXCR3 and CCL17-CCR4 axis are both involved in the pathogenesis of OLP,and mainly play a role in promoting the development of OLP under the influence of local inflammatory microenvironment.