Effects Of CD86 And NF-Κb In Rat Corneal Allograft Immuse Tolerance Inducted By IL-10 Modified Dendritic Cells

ObjectiveObserve the effect of CD86 and NF-Κb in rat corneal allograft immune tolerance inducted by donor IL-10 modified dendritic cells for providing theoretical and practical basis for anti-rejection of corneal transplantation.Methods1. Based on the classical method with GM-CSF plus IL4,part of cells were harvested on the 6th day consist of 6day-imDC.The rest of cells were be derived two groups,one group continue cultivation with IL-4 to the 12th day,the other group we restrained from mature adding IL-10. During the cultivation,we observed the DC form with light microscope and detected the DC phenotypes with flow cytometry.2. Using Wistar-SD rat penetrating corneal transplantation model. The corneal grafts were evaluated by slit- lamp microscope and scored for opacity, edema and vascularization. Survival time of corneal grafts were observed. On the postoperative 14th day, detect the expression of CD86 and NF-κb by the immunohistochemical staining on the corneal grafts of the recipient.Results1. The growth state and form of DCs: The separated cells were spherical, small and floating in the culture fluid. The quantity of cell increased with the culture time lasting,cells'shape in the IL4 group were uniform,sentus getting to cluster, meanwhile sentus in the IL-10 group were not distinct.The purity and phenotype of DCs: greatly significant difference on the CD86 expression level(P<0.01) within three groups indicated that IL-10 restrain DC from mature.2. Survival time of corneal grafts: The mean survival time in the groups of control was merely (10.10±1.66 )days, but in the pretreated groups was (17.80±1.81) days and (26.40±1.65 )days respectively which was statistically prolonged compared with the control groups(P<0.01),and comparison within the two pretreated group is significantly different(P<0.01).Histopathology of corneal allografts: corneas taken from animals of groups B and C had fewer infiltrating cells, an almost normal corneal thickness, and stroma arrangement in order, the center of the grafts showed no inflammatory cellular infiltration. Immunohistochemistry showed that CD86+ and NF-κb+ cells strongly expressed in the control group corneal grafts epithelium.Conclusions1. Abundance of the higher purity immature dendritie cells can be obtained by culturing in vitro with GM-CSF plus IL-4 and restrained DC from mature with IL-10 effectively.2. Derive from donor imDC could markedly prolong the survival time of corneal allografts, relieve acute immune rejection. And IL-10-treated DC can further extending the rat corneal allgrafts. CD86 and NF-κb may play a potential role in the development of corneal graft rejective reaction.