Characterization of swine leukocyte antigen class I genes and homology analysis of the similarity to HLA in GGTA1-/- Wuzhishan minipigs
Received:January 18, 2016  
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KeyWord:GGTA1-/-;Wuzhishan minipigs;Swine leukocyte antigen, SLA;Xenotransplantation;Immune rejection
蒋应弟 甘肃农业大学动物科学技术学院, 兰州 ;中国农业科学院北京畜牧兽医研究所农业部畜禽遗传资源与 种质创新重点实验室, 北京
曾国敏 中国农业科学院北京畜牧兽医研究所农业部畜禽遗传资源与 种质创新重点实验室, 北京 ;甘肃农业大学生命科学技术学院, 兰州
石宁宁 中国农业科学院北京畜牧兽医研究所农业部畜禽遗传资源与 种质创新重点实验室, 北京
李西睿 中国农业科学院北京畜牧兽医研究所农业部畜禽遗传资源与 种质创新重点实验室, 北京
纪慧丽 中国农业科学院北京畜牧兽医研究所农业部畜禽遗传资源与 种质创新重点实验室, 北京
潘登科 中国农业科学院北京畜牧兽医研究所农业部畜禽遗传资源与 种质创新重点实验室, 北京
滚双宝 甘肃农业大学动物科学技术学院, 兰州
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      Objective This study was aimed to characterize the swine leukocyte antigen(SLA) class I genes of GGTA1-/- Wuzhishan minipigs and compare their similarity to human leukocyte antigen(HLA). It has important implications for understanding the cellular rejection in xenotransplantation. Methods Specimens of ear tissue from six founding GGTA1-/- Wuzhishan minipigs were collected, and the SLA class I genes (SLA-1, SLA-3, SLA-2) were amplified by RT-PCR. Purified products were cloned into pEASY-T1 vectors and sequenced, followed by BLAST alignment and using bioinformatc analysis to characterize the SLA class I genes and compare with the similarity to HLA.Results A total of six alleles were detected, among them alleles were previously reported (SLA-1*0703,SLA-2*1102, SLA-3*0401, SLA-3*0403), and the other were novel (SLA-1*0401wz01, SLA-2*11wz01). The homology between alleles of SLA class I genes in Wuzhishan minipigs and HLA was from 70.5% to 72.1%. The homology analysis of critical amino acid residues on HLA binding with human CD8+ molecules showed that SLA-1*0401wz01, SLA-1*0703, SLA-2*11wz01, SLA-2*1102 and SLA-3*0401 occurred mutant at amino acid positions 225 and 228 (T→S,T→M), whereas the other loci were highly conserved. There was a high homology at amino acid level between SLA-2*11wz01, SLA-2*1102 and HLA class I genes which are NK cell KIRs binding sites. Conclusions The amino acid sequences of SLA class I genes of GGTA1-/- Wuzhishan minipigs have a high homology to HLA. From the point of view of cell-mediated xenograft rejection, the amino acid sequences of SLA class I genes of GGTA1-/- Wuzhishan minipigs have a high homology to HLA, therefore, Wzhishan minipigs may become a good potential donor for pig-human xenotransplantation.
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