Evaluation of human and non-human primate antibody binding to pig cells lacking GGTA1/CMAH/β4GalNT2 genes.
Evaluation of human and non-human primate antibody binding to pig cells lacking GGTA1/CMAH/β4GalNT2 genes.
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DOI：
10.1111/xen.12161
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发表时间：
2015-05
影响因子：
3.9
通讯作者：
Tector J
中科院分区：
文献类型：
--
作者：
Estrada JL;Martens G;Li P;Adams A;Newell KA;Ford ML;Butler JR;Sidner R;Tector M;Tector J
猪GGTA 1和CMAH基因的同时失活消除了人抗体识别的碳水化合物异种抗原。β 4GalNT 2糖基转移酶也可以合成异种抗原。为了进一步表征基于聚糖的物种不相容性，我们检查了人和非人灵长类动物抗体与来自缺乏这些碳水化合物修饰基因的转基因猪的细胞的结合。Cas9内切核酸酶和gRNA用于产生缺乏GGTA 1、GGTA 1/CMAH或GGTA 1/CMAH/β 4GalNT 2基因的猪。从这些动物中分离外周血单核细胞，并检查其与人、恒河猴和狒狒IgM和IgG的结合。与缺乏GGTA 1和CMAH的细胞相比，来自GGTA 1/CMAH/β 4GalNT 2缺陷型猪的细胞表现出降低的人IgM和IgG结合。非人灵长类动物抗体与来自各种猪的细胞的反应性表现出与在人类中观察到的反应性略有不同的反应模式。与仅缺乏GGTA 1或缺乏GGTA 1/CMAH/β 4GalNT 2的细胞相比，GGTA 1和CMAH基因的同时失活增加了非人灵长类动物抗体结合。β 4GalNT 2基因的失活降低了人和非人灵长类动物抗体结合，导致猪异种抗原性降低。与缺乏GGTA 1或GGTA 1/CMAH/β 4GalNT 2的猪相比，非人灵长类动物对GGTA 1/CMAH缺陷型细胞的体液免疫增强，突出了碳水化合物异种抗原的复杂性，并表明非人灵长类动物模型在检查某些遗传修饰方面存在潜在局限性。通过猪GGTA 1/CMAH/β 4GalNT 2基因的失活，人免疫球蛋白识别的猪异种抗原逐渐减少，表明异种移植的抗体屏障可以通过基因工程最小化。
Simultaneous inactivation of pig GGTA1 and CMAH genes eliminates carbohydrate xenoantigens recognized by human antibodies. The β4GalNT2 glycosyltransferase may also synthesize xenoantigens. To further characterize glycan-based species incompatibilities, we examined human and non-human primate antibody binding to cells derived from genetically modified pigs lacking these carbohydrate-modifying genes. The Cas9 endonuclease and gRNA were used to create pigs lacking GGTA1, GGTA1/CMAH, or GGTA1/CMAH/β4GalNT2 genes. Peripheral blood mononuclear cells were isolated from these animals and examined for binding to IgM and IgG from humans, rhesus macaques, and baboons. Cells from GGTA1/CMAH/β4GalNT2 deficient pigs exhibited reduced human IgM and IgG binding compared to cells lacking both GGTA1 and CMAH. Nonhuman primate antibody reactivity with cells from the various pigs exhibited a slightly different pattern of reactivity than that seen in humans. Simultaneous inactivation of the GGTA1 and CMAH genes increased nonhuman primate antibody binding compared to cells lacking either GGTA1 only or to those deficient in GGTA1/CMAH/β4GalNT2. Inactivation of the β4GalNT2 gene reduces human and nonhuman primate antibody binding resulting in diminished porcine xenoantigenicity. The increased humoral immunity of nonhuman primates towards GGTA1/CMAH-deficient cells compared to pigs lacking either GGTA1 or GGTA1/CMAH/β4GalNT2 highlights the complexities of carbohydrate xenoantigens and suggests potential limitations of the nonhuman primate model for examining some genetic modifications. The progressive reduction of swine xenoantigens recognized by human immunoglobulin through inactivation of pig GGTA1/CMAH/β4GalNT2 genes demonstrates that the antibody barrier to xenotransplantation can be minimized by genetic engineering.
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DOI：
10.1111/ajt.12744
发表时间：
2014-08
期刊：
American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons
影响因子：
--
作者：
Burlak C;Paris LL;Lutz AJ;Sidner RA;Estrada J;Li P;Tector M;Tector AJ
通讯作者：
Tector AJ
DOI：
10.1073/pnas.1310478110
发表时间：
2013-10-08
影响因子：
11.1
作者：
Tan, Wenfang;Carlson, Daniel F.;Fahrenkrug, Scott C.
通讯作者：
Fahrenkrug, Scott C.
DOI：
10.1089/clo.2006.0079
发表时间：
2007-03-01
期刊：
CLONING AND STEM CELLS
影响因子：
--
作者：
Estrada, Jose;Sommer, Jeffrey;Piedrahita, Jorge A.
通讯作者：
Piedrahita, Jorge A.
影响因子：
3.9
作者：
Padler-Karavani V;Varki A
通讯作者：
Varki A
DOI：
10.4049/jimmunol.1402059
发表时间：
2014-12-01
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Reyes LM;Estrada JL;Wang ZY;Blosser RJ;Smith RF;Sidner RA;Paris LL;Blankenship RL;Ray CN;Miner AC;Tector M;Tector AJ
通讯作者：
Tector AJ