Multiple genetically modified GTKO/hCD46/HLA-E/h2-mg porcine hearts are protected from complement activation and natural killer cell infiltration during ex vivo perfusion with human blood
Multiple genetically modified GTKO/hCD46/HLA-E/h2-mg porcine hearts are protected from complement activation and natural killer cell infiltration during ex vivo perfusion with human blood
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DOI：
10.1111/xen.12390
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发表时间：
2018-09-01
影响因子：
3.9
通讯作者：
Bauer, Andreas
中科院分区：
文献类型：
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作者：
Abicht, Jan-Michael;Sfriso, Riccardo;Bauer, Andreas
在猪-人异种移植中，早期细胞排斥反应是由自然杀伤细胞（NK细胞）介导的。HLA-E通过CD94/NKG2A受体抑制人NK细胞。为了保护猪移植物免受人类NK细胞反应的影响，已经产生了表达hCD46和HLA-E的转基因GTKO猪。本研究的目的是测试这种基因修饰对异种，特别是人类NK细胞反应的影响，使用猪心脏与人类血液的离体灌注模型。方法用新鲜抽取的人血，在离体灌注系统中8小时进行再灌注试验，对移植的转基因（gm）猪心脏（GTKO/hCD46/HLA-E/h2-微球蛋白）和野生型（wt）对照（n=6）进行再灌注试验。在心脏工作模式下，在165分钟的时间内评估心功能。灌注结束时对心肌损伤、抗体沉积、补体活化和凝血参数进行组织学评价。流式细胞术检测灌注液中NK细胞的数量；用免疫荧光显微镜对冷冻切片进行NKp46染色，定量观察NK细胞的组织浸润情况。结果IgG沉积(1.21 × 10(7) vs 8.8 +/- 2.9 × 10(6)；P
BackgroundIn pig-to-human xenotransplantation, early cellular rejection reactions are mediated by natural killer cells (NK cells). Human NK cells are inhibited by HLA-E via CD94/NKG2A receptors. To protect porcine grafts against human NK cell responses, transgenic GTKO pigs expressing hCD46 and HLA-E have been generated. The aim of this study was to test the effect of this genetic modification on xenogeneic, and in particular human NK cell response, using an ex vivo perfusion model of pig hearts with human blood.MethodsCardiopleged and explanted genetically modified (gm) pig hearts (GTKO/hCD46/HLA-E/h2-microglobulin) and wild-type (wt) controls (n=6 each) were reperfused and tested in an 8hours ex vivo perfusion system using freshly drawn human blood. Cardiac function was evaluated during a 165-minute period in working heart mode. Myocardial damage, antibody deposition, complement activation, and coagulation parameters were evaluated histologically at the end of perfusion. The number of NK cells in the perfusate was determined by flow cytometry at baseline and at 8hours; tissue infiltration by NK cells was quantified by immunofluorescence microscopy using NKp46 staining of frozen sections.ResultsDeposition of IgG (1.21x10(7) vs 8.8 +/- 2.9x10(6); P