title: Prevention of rejection of allogeneic endothelial cells in a biohybrid lung by silencing HLA-class I expression.
abstract: 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 β2-microglobulin (β2m)-specific shRNAs reduced β2m 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 TNFα-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.
authors: Department of Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, 30625 Hannover, Germany; Leibniz Research Laboratories for Biotechnology and Artificial Organs, Department for Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, 30625 Hannover, Germany; Lower Saxony Centre for Biomedical Engineering, Implant Research and Development, 30625 Hannover, Germany; German Centre of Lung Research, 30625 Hannover, Germany.
authors: Bettina
authors: B
authors: Wiegmann
authors: Institute for Transfusion Medicine, Hannover Medical School, 30625 Hannover, Germany. Electronic address: Figueiredo.Constanca@mh-hannover.de.
authors: Constança
authors: C
authors: Figueiredo
authors: Institute for Transfusion Medicine, Hannover Medical School, 30625 Hannover, Germany.
authors: Christiane
authors: C
authors: Gras
authors: Leibniz Research Laboratories for Biotechnology and Artificial Organs, Department for Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, 30625 Hannover, Germany.
authors: Michael
authors: M
authors: Pflaum
authors: Leibniz Research Laboratories for Biotechnology and Artificial Organs, Department for Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, 30625 Hannover, Germany.
authors: Sabrina
authors: S
authors: Schmeckebier
authors: Leibniz Research Laboratories for Biotechnology and Artificial Organs, Department for Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, 30625 Hannover, Germany; Lower Saxony Centre for Biomedical Engineering, Implant Research and Development, 30625 Hannover, Germany.
authors: Sotirios
authors: S
authors: Korossis
authors: Department of Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, 30625 Hannover, Germany; Leibniz Research Laboratories for Biotechnology and Artificial Organs, Department for Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, 30625 Hannover, Germany; Lower Saxony Centre for Biomedical Engineering, Implant Research and Development, 30625 Hannover, Germany; German Centre of Lung Research, 30625 Hannover, Germany.
authors: Axel
authors: A
authors: Haverich
authors: Institute for Transfusion Medicine, Hannover Medical School, 30625 Hannover, Germany.
authors: Rainer
authors: R
authors: Blasczyk
mesh_terms: D021522:Bioartificial Organs; D001672:Biocompatible Materials / Q000737:chemistry*; D049109:Cell Proliferation; D016207:Cytokines / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology* / Q000637:transplantation*; D005312:Fetal Blood / Q000166:cytology; D020868:Gene Silencing; D006084:Graft Rejection; D015395:Histocompatibility Antigens Class I / Q000235:genetics*; D006801:Humans; D016040:Lung Transplantation; D010641:Phenotype; D034741:RNA, Small Interfering / Q000378:metabolism; D014184:Transplantation, Homologous; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D001613:beta 2-Microglobulin / Q000008:administration & dosage
chemical_list: D001672:Biocompatible Materials; D016207:Cytokines; D015395:Histocompatibility Antigens Class I; D034741:RNA, Small Interfering; D014409:Tumor Necrosis Factor-alpha; D001613:beta 2-Microglobulin
keywords: Biohybrid lung; Biomaterial; Endothelial cells; HLA