Effect Of LFA-1 Gene Deletion On The Differentiation And Suppressive Function Of CD4~+CD25~+Foxp3~+ Regulatory T Cells Induced By Mice Naive T Cells In Vitro

Background and Aims:To explore the role of LFA-1 gene deletion on the differentiation and suppressive function of CD4+CD25+Foxp3+regulatory T cells induced by mice naive T cell in vitro.Methods:CD4+CD62L+ naive T cells of LFA-1 deficient mice and wild C57/B6 mice (control group) were separated with MACS and the purity was analyzed by flow cytometry(FCM). Naive T cells were cultured in 96-well microplate with bound anti-CD3mAb and anti-CD28 mAb together with soluble murine IL2 and human TGF-β1-1 at 37℃ for 90-108 hours. The ratio of CD4+CD25+Foxp3+ regulatory T was analyzed by FCM. The Foxp3 mRNA of cultured cells was measured by qRT-PCR. All type murine CD4+T cells separated by MACS were stained by CFSE,which were then co-cultured with iTregs in proportion to 1:1.The proliferation index of CD4+ T cells was detected by FCM on 48h-72h.Meanwhile,the cell supernatant was collected for the detection of IL-10 expression.Results:The purity of naive T cells separated by MACS was satisfied for further study. The number of iTregs cells and expression of Foxp3mRNA induced by naive T from LFA-1 deficient mice were lower than that of wild type mice. LFA-1 gene deletion affects differentiation of naive T cells in vitro. Fewer CD4+ CD25+ Foxp3+ regulatory T cells were induced by naive T cells from LFA-1 deletion mice. FCM results shew that LFA-1 gene deletion iTreg cells had stronger inhibition capability than blank control iTreg cells.Howerer,there had no statistical difference between LFA-1 gene deletion group and wild control group iTreg cells. The expression of IL-10 among three groups and comparison between them had statistical differences. IL-10 activity in LFA-1 gene deletion group was highest and the blank control group was lowest.Conclusions:Deficiency of the LFA-1 gene could affect the differentiation of CD4+CD25+Foxp3+ regulatory T cells induced by mice naive T cells in vitro.And IL-10 was involved in this process.Howerer,More evidence are necessary to prove that the repressive effect of LFA-1 deficient iTreg cells to CD4+ T cells has damage.