Genetic engineering of porcine endothelial cell lines for evaluation of human-to-pig xenoreactive immune responses.
Genetic engineering of porcine endothelial cell lines for evaluation of human-to-pig xenoreactive immune responses.
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猪内皮细胞系基因工程用于评估人与猪之间的异种免疫反应。
DOI：
10.1038/s41598-021-92543-y
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发表时间：
2021-06-23
影响因子：
4.6
通讯作者：
Ekser B
中科院分区：
文献类型：
--
作者：
Li P;Walsh JR;Lopez K;Isidan A;Zhang W;Chen AM;Goggins WC;Higgins NG;Liu J;Brutkiewicz RR;Smith LJ;Hara H;Cooper DKC;Ekser B
使用基因工程猪器官的异种移植（跨物种移植）为解决持续的器官短缺提供了一个潜在的解决方案。目前对猪基因修饰的评价主要是监测猪异种器官移植后非人灵长类动物的免疫应答和存活情况。这项措施是临床异种移植试验前的重要步骤，但它耗时，成本高，效率低，变量多。我们开发了一种有效的方法来快速检测体外人对猪的异种免疫应答。猪内皮细胞的特点和永生化的遗传修饰。使用CRISPR/Cas9技术在永生化猪内皮细胞中依次破坏负责产生主要异种抗原（αGal、Neu 5Gc、Sda和SLA-I）的五个基因，包括GGTA 1、CMAH、β 4galNT 2、SLA-I α链和β2-微球蛋白。αGal、Neu 5Gc、Sda和SLA-I的消除显著降低了猪细胞的抗原性，尽管这些细胞仍然保留了其激发人自然杀伤细胞活化的能力。总之，在体外评估人类对转基因猪细胞的免疫应答提供了一种有效的方法来鉴定用于改善猪与人相容性的理想的基因修饰组合，这将加速异种移植到人类的应用。
Xenotransplantation (cross-species transplantation) using genetically-engineered pig organs offers a potential solution to address persistent organ shortage. Current evaluation of porcine genetic modifications is to monitor the nonhuman primate immune response and survival after pig organ xenotransplantation. This measure is an essential step before clinical xenotransplantation trials, but it is time-consuming, costly, and inefficient with many variables. We developed an efficient approach to quickly examine human-to-pig xeno-immune responses in vitro. A porcine endothelial cell was characterized and immortalized for genetic modification. Five genes including GGTA1, CMAH, β4galNT2, SLA-I α chain, and β2-microglobulin that are responsible for the production of major xenoantigens (αGal, Neu5Gc, Sda, and SLA-I) were sequentially disrupted in immortalized porcine endothelial cells using CRISPR/Cas9 technology. The elimination of αGal, Neu5Gc, Sda, and SLA-I dramatically reduced the antigenicity of the porcine cells, though the cells still retained their ability to provoke human natural killer cell activation. In summary, evaluation of human immune responses to genetically modified porcine cells in vitro provides an efficient method to identify ideal combinations of genetic modifications for improving pig-to-human compatibility, which should accelerate the application of xenotransplantation to humans.
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影响因子：
2.2
作者：
Chihara, Ray K.;Lutz, Andrew J.;Tector, A. Joseph
通讯作者：
Tector, A. Joseph
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
影响因子：
3.9
作者：
Fischer, Konrad;Rieblinger, Beate;Schnieke, Angelika
通讯作者：
Schnieke, Angelika
影响因子：
4.4
作者：
Baumann, BC;Forte, P;Seebach, JD
通讯作者：
Seebach, JD
影响因子：
6.2
作者：
Baumann, BC;Schneider, MKJ;Seebach, JD
通讯作者：
Seebach, JD