Prevention of rejection of allogeneic endothelial cells in a biohybrid lung by silencing HLA-class I expression
Prevention of rejection of allogeneic endothelial cells in a biohybrid lung by silencing HLA-class I expression
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DOI：
10.1016/j.biomaterials.2014.06.007
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发表时间：
2014-09-01
期刊：
影响因子：
14
通讯作者：
Blasczyk, Rainer
中科院分区：
文献类型：
--
作者：
Wiegmann, Bettina;Figueiredo, Constanca;Blasczyk, Rainer
人类白细胞抗原（HLA）的变异仍然是异体细胞产品应用的一个障碍。由于生物杂交肺的自体内皮细胞收集不足，异体人脐带血来源的内皮细胞（HCBEC）被用于聚4-甲基-1-戊烯（PMP）气体交换膜的内皮化。因此，HLA I类的表达在HCBECs中被稳定地沉默以防止排斥反应。研究了HLA - i类沉默HCBEC消除异体免疫反应的能力、其功能特性和对PMP膜内皮化的适应性。传递β 2-微球蛋白（β 2m）特异性shrna可将β 2m mRNA水平降低高达90%，并导致HLA I类表达下调高达85%。hla沉默的HCBEC消除了t细胞反应，避免了抗体介导的补体依赖性细胞毒性。表达hla或沉默的HCBEC之间的EC表型和细胞因子分泌谱保持不变。EC特异性激活（如ICAM）和血栓形成标志物（如血栓调节素）不受hla沉默的影响，但它们的表达在TNF α刺激下上调。此外，hla沉默的HCBECs表现出高增殖率，并在PMP膜上构建EC单层。该研究代表了细胞和器官移植领域的一种新的治疗理念，可能使生物人工肺作为肺移植的替代方案更接近现实。(C) 2014 Elsevier Ltd.版权所有。
Variability in Human Leukocyte Antigens (HLA) remains a hurdle to the application of allogeneic cellular products. Due to insufficient autologous endothelial cell harvesting for the biohybrid lung, allogeneic human cord blood derived endothelial cells (HCBEC) were used for the endothelialization of poly-4-methyl-1-pentene (PMP) gas exchange membranes. Therefore, HLA class I expression was silenced stably in HCBECs to prevent rejection. The capacity of HLA class I-silenced HCBEC to abrogate allogeneic immune responses, their functional properties and suitability for endothelialization of PMP membranes were investigated. Delivery of beta 2-microglobulin (beta 2m)-specific shRNAs reduced beta 2m mRNA levels by up to 90% and caused a knockdown of HLA class I expression by up to 85%. HLA-silenced HCBEC abrogated T-cell responses and escaped antibody-mediated complement-dependent cytotoxicity. The EC phenotype and cytokine secretion profiles between HLA-expressing or -silenced HCBEC remained unaltered. EC specific activation (e.g. ICAM) and thrombogenic markers (e.g. thrombomodulin) remained unaffected by HLA-silencing, but their expression was upregulated by TNF alpha-stimulation. Furthermore, HLA-silenced HCBECs showed high proliferation rates and built an EC monolayer onto PMP membranes. This study represents a new therapeutic concept in the field of cell and organ transplantation and may bring the bioartificial lung as an alternative to lung transplantation closer to reality. (C) 2014 Elsevier Ltd. All rights reserved.