Effect Of LncRNA ITGAL On The Differentiation Of CD4~+T Cells Into Treg And Th17 Cells In Corneal Allograft Rejection

Objective(s):To study the effect of lnc RNA ITGAL on the differentiation of CD4~+T cells into Treg and Th17 cells in corneal allograft rejection,and explore the mechanism of corneal allograft rejectionMethods:1.Construct lnc RNA ITGAL overexpression(oe-ITGAL)and small interfering RNA(si-ITGAL)lentiviral vectors,infect normal human peripheral blood CD4~+T cells by cell infection technology,and then perform differentiation induction culture,q PCR to detect the expression level of lnc RNA ITGAL in each group of cells;Flow cytometry detected the differentiation of Th1,Th2,Th17 and Treg,and analyzed the effect of lnc RNA ITGAL on the changes of Treg and Th17cells.2.Construct an allogeneic rat corneal transplantation model,which is divided into control group and intervention group.In the intervention group,lnc RNA ITGAL overexpression and interference with lentiviral subconjunctival injection were used after surgery,and corneal transplantation rejection scores were observed after surgery,and the changes of the number of Treg and Th17 cells in rat whole blood and cervical lymph nodes were measured,and the effect of lnc RNA ITGAL on the changes of Treg and Th17 cells was analyzed.Results:In normal human peripheral blood,the proportion of Th1,Th2,and Th17 cells in the lnc RNA ITGAL overexpression group significantly increased(P<0.0001),while the proportion of Treg cells significantly decreased(P<0.0001);After decreasing the expression of lnc RNA ITGAL,the proportion of Th1,Th2,and Th17 cells significantly decreased(P<0.0001),while the proportion of Treg cells significantly increased(P<0.01).In corneal transplantation rats,there was no significant difference in RI scores between the groups at 7 days and 2 weeks after surgery(P>0.05).In the whole blood of corneal transplantation rats,the proportion of Treg cells and Th17 cells in the lnc RNA ITGAL overexpression group was significantly higher than that in the NC group(P<0.05);The proportion of Treg cells and Th17 cells in the lnc RNA ITGAL interference group was significantly lower than that in the control group NC group(P<0.0001).In the cervical lymph nodes of corneal transplantation rats,the proportion of Treg cells and Th17 cells in the lnc RNA ITGAL overexpression group was significantly higher than that in the NC group(P<0.05);The proportion of Treg cells and Th17 cells in the lnc RNA ITGAL interference group was significantly lower than that in the control group NC group(P<0.05).Conclusion(s):In normal human peripheral blood,the increased expression of lnc RNA ITGAL plays a regulatory role in the differentiation of Th17 and Treg cells,which promotes the differentiation of CD4+T cells to Th17 cells,and inhibits their differentiation to Treg cells,and may participate in the occurrence and development of corneal transplantation rejection through this mechanism.In vivo experiments of corneal transplantation rats,the number of Treg and Th17 cells was positively correlated with the expression level of ITGAL.The expression of lnc RNA ITGAL in the cornea promoted the inflammatory reaction,increased the total number of Treg and Th17 cells in the whole blood and lymph nodes,but did not affect the differentiation of CD4+T cells.