Endothelial cells derived from pigs lacking Galα(1,3)Gal:: no reduction of human leukocyte adhesion and natural killer cell cytotoxicity
Endothelial cells derived from pigs lacking Galα(1,3)Gal:: no reduction of human leukocyte adhesion and natural killer cell cytotoxicity
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DOI：
10.1097/01.tp.0000157231.11083.7c
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发表时间：
2005-05-15
期刊：
影响因子：
6.2
通讯作者：
Seebach, JD
中科院分区：
文献类型：
--
作者：
Baumann, BC;Schneider, MKJ;Seebach, JD
背景半乳糖-α（1，3）半乳糖（Gal）在猪细胞上的表达代表了异种移植的地图或屏障。克服这一障碍的Gal（-/-）猪的产生将研究的重点转向了其他排斥机制，包括细胞免疫。本体外研究调查了（1）人白细胞亚群与来自近交系Ga-/-和Gal（+/+）猪的原代内皮细胞之间的粘附相互作用，以及（2）此类Ga-/-猪内皮细胞对人自然杀伤（NK）细胞毒性的易感性。从Gal（-/-）（PAEC-Gal（-/-））和Ga+/+（PAEC-Gal（+/+））猪分离原代猪主动脉内皮细胞（PAEC）。从健康志愿者中分离人外周血单个核细胞（PBMC）、多形核中性粒细胞（PMN）和NK细胞，并进行功能粘附和细胞毒性试验。人PBMC、PMN或纯化的NK细胞在PAEC-Gal（-/-）细胞上的粘附与在PAEC-Gal（+/+）细胞上的粘附没有差异。比较PBMC的不同白细胞亚群，检测到NK和B细胞对PAEC-Gal（-/-）和PAEC-Gal（+/+）的优先粘附。肿瘤坏死因子-α刺激PAEC-Gal（-/-）和PAEC-Gal（+/+）诱导CD 62 E和CD 106表达增加，并增加细胞粘附，特别是PMN。PAEC-Gal（-/-）细胞上Gal表达的缺乏不能阻止新鲜分离的或白细胞介素-2激活的NK细胞介导的异种人NK细胞的细胞毒性。人白细胞粘附和异种NK细胞对PAEC的细胞毒性均未因缺乏Gal而受损，表明Gal不是细胞排斥的主要靶点。
Background. The expression of galactose-alpha (1,3) galactose (Gal) on porcine cells represents a map or barrier to xenotransplantation. The generation of Gal(-/-) pigs to overcome this barrier redirected the focus of research to other rejection mechanisms, including cellular immunity. The present in vitro study investigated (1) the adhesive interactions between human leukocyte subsets and primary endothelial cells derived from inbred Ga-/- and Gal(+/+) pigs, and (2) the susceptibility of such Ga-/- porcine endothelial cells to human natural killer (NK) cell cytotoxicity.Methods. Primary porcine aortic endothelial cells (PAEC) were isolated from Gal(-/-) (PAEC-Gal(-/-)) and Ga+/+ (PAEC-Gal(+/+)) pigs. Human peripheral blood mononuclear cells (PBMC), polymorphonuclear neutrophils (PMN), and NK cells were isolated from healthy volunteers and tested in functional adhesion and cytotoxicity assays.Results. Adhesion of human PBMC, PMN, or purified NK cells on PAEC-Gal(-/-) cells was not different from that on PAEC-Gal(+/+) cells. Comparing the different leukocyte subsets of PBMC, a preferential adhesion of NK and B cells on both PAEC-Gal(-/-) and PAEC-Gal(+/+) was detected. Tumor-necrosis factor-a stimulation of PAEC-Gal(-/-) and PAEC-Gal(+/+) induced an increase of CD62E and CD106 expression and increased cellular adhesion, in particular, of PMN. The lack of Gal expression on PAEC-Gal(-/-) cells did not prevent xenogeneic human NK-cell cytotoxicity mediated by freshly isolated or interleukin-2-activated NK cells.Conclusions. Neither human leukocyte adhesion nor xenogeneic NK-cell cytotoxicity against PAEC are impaired by the lack of Gal, indicating that Gal is not a dominant target of cellular rejection.