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目的研究转人补体调节蛋白hCD59/hMCP双基因抑制人补体激活从而抑制补体介导的超急性反应、延长体外灌注小鼠心脏功能的作用。方法利用显微注射法建立转hCD59/hMCP双基因小鼠的模型,其子代中hCD59单基因阳性小鼠8只、hMCP单基因阳性小鼠11只和hCD59/hMCP双基因阳性小鼠8只分别为3个实验组,同窝非转基因小鼠10只为对照组。用新鲜人血浆体外灌注小鼠心脏,研究小鼠表达抑制人补体激活的人补体调节蛋白后,对补体介导的异种移植超急性排斥反应的抑制作用。结果转双基因组心脏搏动时间(138min±25min)明显长于单基因组(hCD59组78min±27min,hMCP组43min±21min)及非转基因组(20min±12min)(Dunnett′s T检验,均P<0.01);免疫荧光染色及免疫酶组织化学染色显示转双基因组心脏内补体C3c及C9沉积也明显减少。结论转人补体调节蛋白hCD59/hMCP双基因能够有效地抑制补体介导的异种移植超急性排斥反应。
Objective To study the effect of hCD59 / hMCP double gene transfection on human complement activation, thereby inhibiting complement-mediated hyperacute response and prolonging cardiac perfusion in vitro. Methods The hCD59 double transgenic mouse model was established by microinjection. There were 8 hCD59 single gene positive mice, 11 hMCP single gene positive mice and 8 hCD59 / hMCP double gene positive mice Respectively, three experimental groups, 10 littermate non-transgenic mice as control group. Mouse hearts were infused with fresh human plasma in vitro to study the inhibitory effect of complement inhibition on hyperacute rejection of xenotransplantation in mice expressing inhibitory human complement-activated human complement regulatory proteins. Results The bipedal heart beat time (138min ± 25min) was significantly longer than that of the unicellular group (78min ± 27min in the hCD59 group, 43min ± 21min in the hMCP group) and 20min ± 12min in the non-transgenic group (Dunnett’s T test, all P <0.01) Immunofluorescence staining and immunoenzymatic histochemical staining showed that the deposition of C3c and C9 in the complement of the transgene in the dual genome was also significantly reduced. Conclusion Transfection of hCD59 / hMCP double gene can effectively suppress complement-mediated hyperacute rejection of xenotransplantation.