Exogenous Gene Of IL-10Modified DC Induced Immune Tolerance On Cornea Transplant In Rats

Objective1、 To investigate the phenotypes and biological characteristics ofinterleukin-10gene modified dendritic cell(DC).2、By establishing rat penetrating keratoplasty model,we research aboutexogenous gene of IL-10modified DC induced immune tolerance on corneatransplant in rats.Methods1、The DCs were trained six days,after which were randomly divided intofour groups,the GFP-interleukin-10gene modified dendritic cell(IL-10-GFP-DC), the GFP gene modified dendritic cell(GFP-DC),the eighthdayt-DC(8-DC)，the twelfth day-DC(12-DC). the protein of IL-10was detected byWestern Blot,the flow cytometry was used to detect the phenotypes,and theproliferation experiment shows the proliferation activity of each group.2、Animal experiments were divided into five groups: Negative control group,Positive control group,GFP-DC group,8-DCgroup and IL-10-GFP-DC group.Wecollect cells of GFP-DC,8-DC and IL-10-GFP-DC from donor Wistar rats,andwhith were injected into receptor SD rats though tail vein2×106cell, thepositive control group given the same amount PBS.After three days, ratpenetrating keratoplasty model would be established. we observed thecondition of corneal plants, recording the survival time and pathologicalchanges.the change of IL-10、TGF-βand IL-2would be detected by RT-PCR,ELISA was used to detect the change of IL-4and IFN-γ. Result1、The result of Western Blot show that IL-10could be the most effectiveexpressed in IL-10-GFP-DC,The phenotypes of CD83and CD86were theLowest expression in IL-10-GFP-DC in three groups and the proliferation activityof allogeneical T cell was the same lowly.2、The corneal plants survival time of IL-10-GFP-DC group is the longest andlittle inflammatory reaction,the expression of IL-10、TGF-βand IL-4weremuch more than the others,and IL-2and IFN-γ were lowest expression ofIL-10-GFP-DC group.Conclusion1、The modified by IL-10gene could inhibit the mature dendritic cells,andmaking basic of rat penetrating keratoplasty model.2、Immature dendritic cells inhibit corneal transplantion rejection, prolong thecorneal plants survival time. the ratio imbalance of Th1/Th2and activation ofTGF-βplayed an important role in immune tolerance.