五指山小型猪-藏酋猴异种肝移植的术前检测

摘要

目的 本研究为了减少五指山小型猪-藏酋猴异种肝移植手术的超急性排斥反应(hyperacute rejection,HAR)和急性排斥反应(acute rejection,AR)发生率,进行全面术前检测.方法 分别利用PCR技术和植物凝集素细胞染色对供体GGTA1基因敲除类型进行鉴定;30只藏酋猴分别与五指山小型猪进行淋巴细胞毒实验;供受体ABO血型鉴定.结果 供体为GGTA1基因敲除猪,GTKO猪外周血单个核细胞(peripheral blood mononuclear cell,PBMC)植物凝集素细胞染色未检测出荧光,表明GGTA1基因敲除成功,与基因型鉴定结果一致.通过淋巴细胞毒实验选定28#与供体PBMC毒性最小的藏酋猴做为备选受体.血型鉴定显示供受体血型一致.结论 通过对GTKO五指山小型猪-藏酋猴异种肝移植的术前检测选定合适的受体,可有效减少异种移植HAR和AR的发生率.%Objective To perform comprehensive preoperative testing in order to reduce the risk of hyperacute rejection (HAR) and acute rejection(AR) in the Wuzhishan miniature pig-Tibetan monkey xenotransplantation. To Minimize the influence of immunological factors on recipients survival in order to prolong the recipients survival. Methods The donor GGTA1 gene knockout type was identified by PCR and lectin cell staining, respectively. A total number of 30 Tibetan monkey and Wuzhishan miniature pigs were tested for lymphocyte toxicity and ABO blood group identification. Results The donor was GGTA1 knockout pigs, and the fluorescence was not detected in GTKO pig peripheral blood mononuclear cell(PBMC)with lectin cell staining, it indicated that the GGTA1 gene was successfully knocked out,which was consistent with the genotype identification results. The Tibetan monarch monkey 28# with the least toxicity to the donor PBMC through lymphocyte toxicity test was selected as a candidate recipient. Blood type test showed identical blood type between donor and recipients. Conclusion The preoperative test of GTKO Wuzhishan miniature pigs-Tibetan monkey xenotransplantation has greatly reduced the risk of HAR and AR after xenotransplantation.