Antigenicity of tissues and organs from GGTA1/CMAH/β4GalNT2 triple gene knockout pigs
Ronggen Wang, Miaomiao Ruan, Runjie Zhang, Lei Chen, Xiaoxue Li, Bin Fang, Chu Li, Xueyang Ren, Jiying Liu, Qiang Xiong, Lining Zhang, Yong Jin, Lin Li, Rongfeng Li, Ying Wang, Haiyuan Yang, Yifan Dai
Antigenicity of tissues and organs from GGTA1/CMAH/β4GalNT2 triple gene knockout pigs
Clinical xenotransplantations have been hampered by human preformed antibody-mediated damage of the xenografts. To overcome biological incompatibility between pigs and humans, one strategy is to remove the major antigens [Gal, Neu5Gc, and Sd(a)] present on pig cells and tissues. Triple gene (GGTA1, CMAH, and b4GalNT2) knockout (TKO) pigs were produced in our laboratory by CRISPR-Cas9 targeting. To investigate the antigenicity reduction in the TKO pigs, the expression levels of these three xenoantigens in the cornea, heart, liver, spleen, lung, kidney, and pancreas tissues were examined. The level of human IgG/IgM binding to those tissues was also investigated, with wildtype pig tissues as control. The results showed that αGal, Neu5Gc, and Sd(a) were markedly positive in all the examined tissues in wildtype pigs but barely detected in TKO pigs. Compared to wildtype pigs, the liver, spleen, and pancreas of TKO pigs showed comparable levels of human IgG and IgM binding, whereas corneas, heart, lung, and kidney of TKO pigs exhibited significantly reduced human IgG and IgM binding. These results indicate that the antigenicity of TKO pig is significantly reduced and the remaining xenoantigens on porcine tissues can be eliminated via a gene targeting approach.
pig / xenotransplantation / GGTA1 / CMAH / β4GalNT2 / antigenicity
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