人CD59/MCP双基因对人血灌注转基因小鼠离体心脏功能的保护作用

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。

Coexpression of human complement regulatory protein genes, CD59 and MCP in transgenic mice protects isolated mouse heart function in ex vivo human plasma perfusion model

OBJECTIVE: To investigate if the expression of human complement regulatory protein genes in transgenic donor protects against hyperacute rejection (HAR) in the recipient. METHODS: Kunming mice were transfected with 2 human complement regulatory protein genes, CD59 and MCP, so as to establish a transgenic hCD59/hMCP mouse model. Eight hCD59 expression mice, 11 hMCP expression mice, and 8 hCD59/hMCP expression mice were used as experimental groups, and 10 transgenic negative littermates were used as control group. The hearts of the mice were taken out to be perfused with 10% pooled human blood of B type. During the perfusion electrocardiography was carried out to observe the beating time. After the hearts stopped beating, immunofluorescence staining and immunohistochemistry were used to detect the deposition of complements C(9) and C(3c) in the heart tissue. RESULTS: The mean heart beating time was 138 +/- 25 minutes in the hCD59/hMCP expression group, 78 +/- 27 minutes in the hCD59 expression group, 43 +/- 21 minutes in the hMCP group, and 20 +/- 12 minutes in the wild type nontransgenic control group (all P < 0.01, Dennett's T test for all 3 other groups relative to the hCD59/hMCP group). Deposition of the complement C(9) and that of C(3c) were not found in the hearts of the hCD59/hMCP group, however, could be found in the hearts of the 2 monotransgenic groups and nontransgenic group. CONCLUSION: The coexpression of human complement regulatory protein genes, CD59 and MCP in the xenografts effectively inhibits the complement of xenograft-mediated HAR.