• ISSN 1674-8301
  • CN 32-1810/R
Volume 33 Issue 4
Jul.  2019
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Article Contents
Wang Ronggen, Ruan Miaomiao, Zhang Runjie, Chen Lei, Li Xiaoxue, Fang Bin, Li Chu, Ren Xueyang, Liu Jiying, Xiong Qiang, Zhang Lining, Jin Yong, Li Lin, Li Rongfeng, Wang Ying, Yang Haiyuan, Dai Yifan. Antigenicity of tissues and organs from GGTA1/CMAH/β4GalNT2 triple gene knockout pigs[J]. The Journal of Biomedical Research, 2019, 33(4): 235-243. doi: 10.7555/JBR.32.20180018
Citation: Wang Ronggen, Ruan Miaomiao, Zhang Runjie, Chen Lei, Li Xiaoxue, Fang Bin, Li Chu, Ren Xueyang, Liu Jiying, Xiong Qiang, Zhang Lining, Jin Yong, Li Lin, Li Rongfeng, Wang Ying, Yang Haiyuan, Dai Yifan. Antigenicity of tissues and organs from GGTA1/CMAH/β4GalNT2 triple gene knockout pigs[J]. The Journal of Biomedical Research, 2019, 33(4): 235-243. doi: 10.7555/JBR.32.20180018

Antigenicity of tissues and organs from GGTA1/CMAH/β4GalNT2 triple gene knockout pigs

doi: 10.7555/JBR.32.20180018
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  • Corresponding author: Haiyuan Yang and Yifan Dai, Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, 101 Longmian Avenue, Nanjing, Jiangsu 211166, China. E-mails: hyyang@njmu.edu.cn and daiyifan@njmu.edu.cn
  • Received: 2018-02-12
  • Revised: 2018-03-30
  • Accepted: 2018-04-23
  • Published: 2018-05-31
  • Issue Date: 2019-07-01
  • 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 β4GalNT2) 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.

     

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