Effect Of Cytokines And Serial Subcultivation On The Expressions Of HLA-ABC,HLA-DR And ICAM-1 Molecules Of Human Retinal Pigment Epithelium In Vitro

Objective To investigate the effect of cytokine and serial subcultivation on theimmunological competence of human retinal pigment epithelium (hRPE) in vitro.Methods The immunological competence of hRPE was observed by flow cytometry todetect the expressions of HLA-ABC,HLA-DR and ICAM-1 molecules.Result Normal hRPE cells expressed high levels of HLA-ABC molecules and low levels ofHLA-DR and ICAM-1 molecules. Stimulated hRPE cells were obtained by incubation withrecombinant human interferon-γ(rhIFN-γ), transforming growth factor-β1 (TGF-β1),transforming growth factor-β2 (TGF-β2)and interleukin-10 (IL-10). The expressions ofHLA-ABC, HLA-DR and ICAM-1 molecules were strongly up-regulated in the presence ofrhIFN-γ(P<0.001). TGF-β1 and TGF-β2 did not suppress the hRPE's expression ofHLA-ABC, HLA-DR and ICAM-1 molecules without the IFN-γstimulation (P>0.05). Butthey both suppressed the rhIFN-γinduced up-regulation of HLA-DR molecule (P<0.001) andTGF-β2 suppressed the rhIFN-γinduced up-regulation of HLA-ABC molecules (P=0.48).IL-10 did not suppress the hRPE's expression of HLA-ABC, HLA-DR and ICAM-1molecules without the IFN-γstimulation (P>0.05). But it suppressed the rhIFN-γinducedup-regulation of HLA-DR molecule (P<0.001).The duration of culture in vitro weresignificantly correlated with.the expression of HLA-ABC molecules (The rate of positive cell:r = -0.924,P <0.001 , The mean fluorescence intensity: r = -0.820,P <0.001 ), andICAM-1 molecule (The rate of positive cell: r = 0.914,P <0.001 , The mean fluorescenceintensity: r = 0.828,P <0.001). And no correlations between the duration of culture in vitroand the expressions of HLA-DR molecule (The rate of positive cell: r = 0.106,P = 0.356,The mean fluorescence intensity: r = -0.007,P = 0.948) were observed..Conclussion IFN-γstrengthens the antigenicity of hRPE, which plays a role in theimmunological rejection after hRPE transplantion. TGF-β1, TGF-β2 and IL-10 weaken thatof hRPE, which plays a role in avoiding the immunological rejection after hRPEtransplantion. Chronic rejection is the main type of graft rejective reaction post hRPEtransplantation. The rate of acute rejection is down-regulated and that of chronic rejection isup-regulated when the passaging of hRPE cells are used for transplantation.