The Effect Of LFA-1 On Plasticity Of Treg In Inflammation Conditions In Vitro

Background and Aims:In recent years,the plasticity of CD4+T cells was researched entensively,especially in Treg cells and Th17 cells on adaptive immune response to different internal environment changing.Lymphocyte function associated antigen 1(LFA-1)has important role on the function and migration of lymphocyte differentiation by providing coordinated stimulus receptors and signal to affect lymphocyte cells' activation,proliferation and differentiation.The study was to observe the effect and mechanism of LFA-1 on the function and plasticity of Treg cells in different inflammatory conditions.Methods:Collecting PBMC cells were collected in spleen from wild type and LFA-1-/-mice and the naive T cells were purified by the magnetic bead separation method.Naive T cells was directional differentiation to Treg induced by cytokine IL-2,TGF-beta in vitro for 5 days and then the expresion of Foxp3 measured by flow cytometry.Meanwhile,the directional differentiation cells were separated by magnetic beads to obtain high purity of Treg cells.The above two kinds of cells were placed in the complete culture medium containing IL-6,IL-23 and TGF-? in the simulated inflammatory environment.On the 8th day,the expression of IL-17 and Foxp3 was detected by flow cytometry,and the expression of STAT3 and ROR gamma t was detected by PCR.The levels of IL-10,IL-17 in supernatant were measured by ELSIA.Result:Both of the wild mice Treg cells and LFA-1-/-mice Treg cells were shown the plasticity in inflammatory conditions,but LFA-1-/-mice Treg cells revealed a higher percentage of Foxp3/IL-17 co-expressing5 and the expression of STAT3 and ROR gamma t in LFA-1-/-mice Treg was also increased than that in wild mice Treg cells.Furthermore,both of the two types cell supernatants were shown that IL-17 is higher with IL-10 decreasing than that of treatment before,and the change is obvious in LFA-1-/-mice Treg cells.Conclusions:Inflammatory factors in inflammatory environments can mediate the transformation of functional plasticity of Treg cells,which may increase the production of pathogenic factor IL-17 by increasing the expression of IL-17 and Foxp3 and reducing the production of anti-inflammatory factor IL-1O.The deletion of LFA-1 gene makes the change of functional plasticity of Treg is more obvious.