Related Research Of The Expression Of Transcription Factors GATA-3,p-STAT6 And P-STAT3 In Intermingled Skin Transplantation Induced By B7-H1 Expressed On Auto-keratinocytes

ObjectiveThis research aim to study the expression of the transcription factors GATA-3,p-STAT6 and p-STAT3 in intermingled skin transplantation and the relationship between GATA-3,p-STAT6 and p-STAT3 in lymphocytes and B7-H1 expressed on auto-keratinocytes in intermingled skin transplantation,and to explore the mechanism of the local immuno-suppression in intermingled skin transplantation.Methods 1.lymphocytes,epidermic cells and keratinocytes were seperated from suckling mice and then identifyied the keratinocytes by immunohistochemistry.2.Mixed culture of lymphocytes,epidermic cells and keratinocytes in vitro and established the model of intermingled skin grafting.3.After 24 h,Isolated keratinocytes and lymphocytes in MELC.Western-blot was performed to detect the expressions of B7-H1 in MELC.The expressions of GATA-3,STAT6,STAT3 and IL-10 mRNA in lymphocytes were detected by Real-time quantitative PCR.We did the same operation after 48 h.4.Added the anti-B7-H1 monclonal antibody to the MELC with auto-keratinocytes and detected the expressions of GATA-3,STAT6,STAT3 and IL-10 mRNA in lymphocytes by Real-time quantitative PCR.Results 1.The activity of three Primary cells was good and the culture was stable in vitro.The immunohistochemistry indicated the cells could express PKC and confirmed the cells were keratinocytes.2.Through Western-blot,in the MELC with auto-keratinocytes,the expression of B7-H1 were rising trend in both 24 h and 48 h.But the expression of B7-H1 did not have the trend in MELC without auto-keratinocytes,3.Compared with the comtrol groups without auto-keratinocytes,RT-PCR assay indicated that the relative levels of GATA-3,STAT6,STAT3 andIL-10 mRNA expression were significant raised in both 24 h and 48h(P<0.05).4.Compared with the MELC without anti-B7-H1 monclonal antibody and theMELC withcontrol antibody,the expression of GATA-3,STAT6,STAT3andIL-10 mRNA in the MELC with anti-B7-H1 monclonal antibody were significant decreased in both 24 h and 48h(P<0.05).ConclusionIn the intermingled skin grafting model,the auto-keratinocytes could raise the expression of GATA-3,p-STAT6,p-STAT3 and IL-10 in lymphocytes and it depended on B7-H1.It could suppressed the immune response in the intermingled skin grafting model.