Study About Effects Of Il-10-treated Dendritic Cells On Rat Corneal Transplantation

Objective1.The purpose of this section was to study the changes of interleukin-10-treated dendritic cells in phenotype and function.2.By establishing stable rat penetrating keratoplasty model, we looked for new methods inhibiting corneal transplantation rejection.3.Through RT-PCR and pathological methods, we detected the expressions of cytokines and inflammatory reactions and discussed corneal immune tolerance mechanism induced by immature dendritic cells.Methods1.With lymphocyte separation medium of rat, we extracted precursors of dendritic cells from rat bone marrow. We started cell primary culture by induction of cytokines. At the sixth day of the culture, the experiment was divided into three groups: the sixth day-DC(6-DC); the twelfth day-DC(12-DC); the IL-10-treated-DC(IL-10-DC). We compared differences in morphology, phenotype and function among the three groups.2.Trials were divided into three groups: the positive control group, 6-DC group, IL-10-DC group, eight one group.we collected 6-DC, IL-10-DC from Wistar rats, which were given to receptor SD rats by tail intravenous per 2 x 106cells and the positive control group given PBS. After three days, we established penetrating corneal transplantation model. Every day we observed the postoperative corneal allografts and recorded the survival time.3.After operation, we took out 6 rats respectively to detect the expressions of cell factors IL-12, IL-2, IFN-gamma,IL-4 and IL-10 by RT-PCR on the 14th day. The rest were used to check inflammatory responses by histopathological examination.Results1.Dentritic cells had typical morphologies. The results of the flow cytometry revealed that there were significantly statistic differences among the three groups. The expression rates of CD86 and CD83 were the highest in 12-DC and the lowest in IL-10-DC. MTT displayed the same results.2.Survival time between 6-DC group and IL-10-DC group was apparently prolonged.There were statistically significant differences.3. The results of RT-PCR indicated Th1 cell factors and IL-12 were obvious in the positive control group and high expressions of Th2 cytokines in 6-DC group and IL-10-DC group. In the positive control group, we observed corneal epithelial edema, the stromal layer fiber structure disorder, inflammatory cells infiltrating and visible new blood vessels. 6-DC group and IL-10-DC group showed few inflammatory reactions.Conclusions1.We successfully cultivated dendritic cells. IL-10 could inhibit the maturities of dendritic cells.2.Immature dendritic cells could inhibit the corneal transplantation rejection. The effect of IL-10-DC inducing immune tolerance was much better.3.Th1 cell factors and IL-12 were related to the corneal transplantation rejection, Th2 cytokines related to immunodepression. IL-10-DC could inhibit inflammatory reactions of cornea.