Transgenic expression of human leukocyte antigen-E attenuates GalKO.hCD46 porcine lung xenograft injury
| Autor: | Xiangfei Cheng, Richard N. Pierson, Natalia Kubicki, Arielle Cimeno, Gheorghe Braileanu, Carol Phelps, Lars Burdorf, Beth M. French, Andrea Bähr, Christopher Laird, Edward So, David Ayares, Dawn Parsell, Wenji Sun, Natalie A. O'Neill, Nikolai Klymiuk, Agnes M. Azimzadeh |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Cytotoxicity Immunologic Pathology medicine.medical_specialty Endothelium Swine Immunology Cell Transplantation Heterologous Hemodynamics Inflammation 030230 surgery Biology Andrology Animals Genetically Modified Membrane Cofactor Protein 03 medical and health sciences 0302 clinical medicine HLA Antigens medicine Leukocytes Animals Humans Platelet activation Cell adhesion Transplantation Lung Graft Survival Endothelial Cells Lung Injury Galactosyltransferases Killer Cells Natural 030104 developmental biology medicine.anatomical_structure Heterografts medicine.symptom Ex vivo |
| Zdroj: | Xenotransplantation. 24(2) |
| ISSN: | 1399-3089 |
| Popis: | Background: Lung xenografts remain susceptible to loss of vascular barrier function within hours in spite of significant incremental advances based on genetic engineering to remove the Gal 1,3-Gal antigen (GalTKO) and express human membrane cofactor protein (hCD46). Natural killer cells rapidly disappear from the blood during perfusion of GalTKO.hCD46 porcine lungs with human blood and presumably are sequestered within the lung vasculature. Here we asked whether porcine expression of the human NK cell inhibitory ligand HLA-E and 2 microglobulin inhibits GalTKO.hCD46 pig cell injury or prolongs lung function in two preclinical perfusion models. Methods: Lungs from pigs modified to express GalTKO.hCD46 (n=37) and GalTKO.hCD46.HLA-E (n=5) were harvested and perfused with human blood until failure or elective termination at 4hours. Airway pressures and pulmonary artery hemodynamics were recorded in real time. Blood samples were also collected throughout the experiment for analysis. Porcine aortic endothelial cells (PAECs) from each genotype were cultured in monolayers in microfluidic channels and used in fluorescent cytotoxicity assays using human NK cells. Results: HLA-E expression on GalTKO.hCD46 PAECs was associated with significantly decreased antibody-dependent and antibody-independent NK-mediated cytotoxicity under in vitro conditions simulating physiologic shear stress. Relative to GalTKO.hCD46 pig lungs perfused with human blood on an ex vivo platform, additional expression of HLA-E increased median lung survival (>4hours, vs 162minutes, P=.012), and was associated with attenuated rise in pulmonary vascular resistance, and decreased platelet activation and histamine elaboration. As expected, HLA-E expression was not associated with a significant difference in NK cell adhesion to endothelial cells in vitro, or NK cell and neutrophil sequestration during organ perfusion. Conclusions: We conclude human NK cell activation contributes significantly to GalTKO.hCD46 pig endothelial injury and lung inflammation and show that expression of HLA-E is associated with physiologically meaningful protection of GalTKO.hCD46 cells and organs exposed to human blood. |
| Databáze: | OpenAIRE |
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