Immunogenicity of Renal Microvascular Endothelial Cells From Genetically Modified Pigs
| Autor: | Zheng Yu Wang, Jessica B. Montgomery, Jose L. Estrada, A. Joseph Tector, Ross L. Blankenship, Ping Li, Matthew Tector, Shunji Nagai, Susan M. Downey, James R. Butler |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Antigenicity Glycan Swine 030230 surgery Kidney Transfection Peripheral blood mononuclear cell Mixed Function Oxygenases Flow cytometry Animals Genetically Modified Gene Knockout Techniques 03 medical and health sciences 0302 clinical medicine Antigens Heterophile medicine Animals Humans Gene silencing Cells Cultured Transplantation medicine.diagnostic_test biology Chemistry Immunogenicity Graft Survival Endothelial Cells Galactosyltransferases Molecular biology In vitro Phenotype 030104 developmental biology Immunoglobulin M Immunoglobulin G Microvessels biology.protein N-Acetylgalactosaminyltransferases Antibody |
| Zdroj: | Transplantation. 100:533-537 |
| ISSN: | 0041-1337 |
| Popis: | BACKGROUND Disrupting the porcine GGTA1 and CMAH genes [double knockout (DKO)] that produce the gal-α(1,3)-gal and N-glycolylneuraminic acid xenoantigens reduces human antibody binding to porcine peripheral blood mononuclear cells. It is important to examine rejection pathways at an organ-specific level. The object of this study is to evaluate the human preformed antibody reactivity against DKO renal microvascular endothelial cells (RMEC) in vitro. METHODS Characteristics of DKO RMEC were analyzed using flow cytometry. Human IgG/M binding to primary RMEC, immortalized RMEC (iRMEC), and iRMEC-deficient in B4GALNT2 genes were examined using flow cytometric crossmatch assay. RESULTS Porcine RMEC expressed gal-α(1,3)-gal, N-glycolylneuraminic acid, and Dolichos biflorus agglutinin glycans recognized by human preexisting antibodies in humans. Antigenicity of DKO RMEC was lower than GGTA1 KO RMEC. The disruption of B4GALNT2 gene in DKO iRMEC further reduced human IgG/IgM binding. CONCLUSIONS Silencing the porcine GGTA1, CMAH, and B4GALNT2 genes is an effective strategy to reduce human preformed antibody binding to RMEC. Porcine RMEC will be a useful reagent for the further study of xenoimmunology. |
| Databáze: | OpenAIRE |
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