In vitro investigation of pig cells for resistance to human antibody-mediated rejection
In vitro investigation of pig cells for resistance to human antibody-mediated rejection
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DOI：
10.1111/j.1432-2277.2008.00736.x
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发表时间：
2008-12-01
影响因子：
3.1
通讯作者：
Cooper, David K. C.
中科院分区：
文献类型：
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作者：
Hara, Hidetaka;Long, Cassandra;Cooper, David K. C.
尽管α 1，3-半乳糖基转移酶基因敲除（GTKO）猪细胞的人补体依赖性细胞毒性（CDC）明显弱于野生型（WT）细胞，但成功的异种移植将需要具有多种遗传修饰的猪。通过（i）流式细胞术检测来自健康人的血清的IgM/IgG结合，和（ii）针对来自五种类型的猪的外周血单核细胞和猪主动脉内皮细胞的CDC测定-WT、GTKO、H-转移酶转基因的GTKO（GTKO/HT）、人补体调节蛋白CD46转基因的WT（CD46）和GTKO/CD46。与WT和CD 46细胞的平均IgM/IgG结合显著高于与GTKO、GTKO/HT和GTKO/CD 46的平均IgM/IgG结合，但GTKO、GTKO/HT和GTKO/CD 46细胞之间无差异。WT的平均CDC显著高于GTKO、GTKO/HT、CD 46和GTKO/CD 46细胞，但GTKO和GTKO/HT之间无差异。GTKO/CD 46细胞的裂解率明显低于GTKO或CD 46细胞。CD46表达提供了对来自对GTKO猪心脏致敏的狒狒的血清的部分保护。GTKO/CD 46细胞对人血清和致敏狒狒血清的裂解具有显著抵抗性。总之，通过Gal表达的缺失和CD46表达的存在的组合获得了对CDC的最大保护，但是HT的表达似乎没有提供优于GTKO的优势。来自GTKO/CD46猪的器官可能对CDC的敏感性显著降低。
Although human complement-dependent cytotoxicity (CDC) of alpha 1,3-galactosyltransferase gene-knockout (GTKO) pig cells is significantly weaker than that of wild-type (WT) cells, successful xenotransplantation will require pigs with multiple genetic modifications. Sera from healthy humans were tested by (i) flow cytometry for binding of IgM/IgG, and (ii) CDC assay against peripheral blood mononuclear cells and porcine aortic endothelial cells from five types of pig - WT, GTKO, GTKO transgenic for H-transferase (GTKO/HT), WT transgenic for human complement regulatory protein CD46 (CD46) and GTKO/CD46. There was significantly higher mean IgM/IgG binding to WT and CD46 cells than to GTKO, GTKO/HT, and GTKO/CD46, but no difference between GTKO, GTKO/HT, and GTKO/CD46 cells. There was significantly higher mean CDC to WT than to GTKO, GTKO/HT, CD46, and GTKO/CD46 cells, but no difference between GTKO and GTKO/HT. Lysis of GTKO/CD46 cells was significantly lower than that of GTKO or CD46 cells. CD46 expression provided partial protection against serum from a baboon sensitized to a GTKO pig heart. GTKO/CD46 cells were significantly resistant to lysis by human serum and sensitized baboon serum. In conclusion, the greatest protection from CDC was obtained by the combination of an absence of Gal expression and the presence of CD46 expression, but the expression of HT appeared to offer no advantage over GTKO. Organs from GTKO/CD46 pigs are likely to be significantly less susceptible to CDC.