双重敲除 N-羟乙酰神经氨酸和半乳糖 α-1,3-半乳糖的猪降低了异种移植的体液屏障。
Double knockout pigs deficient in N-glycolylneuraminic acid and galactose α-1,3-galactose reduce the humoral barrier to xenotransplantation.
机构信息
Department of Surgery, IU Health Transplant Institute, Indiana University School of Medicine, Indianapolis, IN 46202, USA.
出版信息
Xenotransplantation. 2013 Jan-Feb;20(1):27-35. doi: 10.1111/xen.12019.
BACKGROUND
Clinical xenotransplantation is not possible because humans possess antibodies that recognize antigens on the surface of pig cells. Galα-1,3-Gal (Gal) and N-glycolylneuraminic acid (Neu5Gc) are two known xenoantigens.
METHODS
We report the homozygous disruption of the α1, 3-galactosyltransferase (GGTA1) and the cytidine monophosphate-N-acetylneuraminic acid hydroxylase (CMAH) genes in liver-derived female pig cells using zinc-finger nucleases (ZFNs). Somatic cell nuclear transfer (SCNT) was used to produce healthy cloned piglets from the genetically modified liver cells. Antibody-binding and antibody-mediated complement-dependent cytotoxicity assays were used to examine the immunoreactivity of pig cells deficient in Neu5Gc and Gal.
RESULTS
This approach enabled rapid production of a pig strain deficient in multiple genes without extensive breeding protocols. Immune recognition studies showed that pigs lacking both CMAH and GGTA1 gene activities reduce the humoral barrier to xenotransplantation, further than pigs lacking only GGTA1.
CONCLUSIONS
This technology will accelerate the development of pigs for xenotransplantation research.
背景
临床异种移植是不可能的，因为人类具有识别猪细胞表面抗原的抗体。Galα-1,3-Gal（Gal）和 N- 糖基神经氨酸（Neu5Gc）是两种已知的异种抗原。
方法
我们使用锌指核酸酶（ZFNs）报告了在肝源性雌性猪细胞中同源敲除 α1,3-半乳糖基转移酶（GGTA1）和胞苷单磷酸-N-乙酰神经氨酸羟化酶（CMAH）基因。体细胞细胞核移植（SCNT）用于从基因修饰的肝细胞中产生健康的克隆猪。抗体结合和抗体介导的补体依赖性细胞毒性试验用于检测缺乏 Neu5Gc 和 Gal 的猪细胞的免疫反应性。
结果
这种方法能够快速生产缺乏多个基因的猪品系，而无需广泛的繁殖方案。免疫识别研究表明，缺乏 CMAH 和 GGTA1 基因活性的猪降低了异种移植的体液屏障，比仅缺乏 GGTA1 的猪更有效。
结论
这项技术将加速用于异种移植研究的猪的开发。
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