Faculty Information |
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Article types | Original article |
Language | English |
Refereed paper | Refereed |
Title | Human HLA-Ev (147) Expression in Transgenic Animals |
Journal | Formal name:Transplantation proceedings Abbreviation:Transplant Proc ISSN code:1873-2623(Electronic)0041-1345(Linking) |
Domestic / Foregin | Foregin |
Volume, Number, Page | 48(4),pp.1323-5 |
Papers・Author | Matsuura R, Maeda A, Sakai R, Eguchi H, Lo P-C, Hasuwa H, Ikawa M, Nakahata K, Zenitani M, Yamamichi T, Umeda S, Deguchi K, Okuyama H, Miyagawa S |
Publication date | 2016/05 |
Papers・Description | BACKGROUND:In our previous study, we reported on the development of substituting S147C for HLA-E as a useful gene tool for xenotransplantation. In this study we exchanged the codon of HLA-Ev (147), checked its function, and established a line of transgenic mice.METHODS:A new construct, a codon exchanging human HLA-Ev (147) + IRES + human beta 2-microgloblin, was established. The construct was subcloned into pCXN2 (the chick beta-actin promoter and cytomegalovirus enhancer) vector. Natural killer cell- and macrophage-mediated cytotoxicities were performed using the established the pig endothelial cell (PEC) line with the new gene. Transgenic mice with it were next produced using a micro-injection method.RESULTS:The expression of the molecule on PECs was confirmed by the transfection of the plasmid. The established molecules on PECs functioned well in regulating natural killer cell-mediated cytotoxicity and macrophage-mediated cytotoxicity. We have also successfully generated several lines of transgenic mice with this plasmid. The expression of HLA-Ev (147) in each mouse organ was confirmed by assessing the mRNA. The chick beta-actin promoter and cytomegalovirus enhancer resulted in a relatively broad expression of the gene in each organ, and a strong expression in the cases of the heart and lung.CONCLUSION:A synthetic HLA-Ev (147) gene with a codon usage optimized to a mammalian system represents a critical factor in the development of transgenic animals for xenotransplantation. |
DOI | 10.1016/j.transproceed.2015.10.069 |
PMID | 27320614 |