| Autor: |
Park, Kwang Wook, Kim, Eun Jung, Han, Gui Seung, Choi, Ki Myung, Hong, Seung Pyo, Yoo, Ji Yeon, Kim, Soo Hyun, Park, Young Chul, Seol, Jae Goo |
| Zdroj: |
Biology of Reproduction; May 2008, Vol. 78 Issue: 1, Number 1 Supplement 1 p293-293, 1p |
| Abstrakt: |
The pig has been identified as a suitable organ donor for humans. However multiple immune barriers preclude clinical applications of pig's organ. In particular, the severe type of rejection in xenotransplantation is cellular rejection by host CD8+ cytotoxic T lymphocyte (CTL) cells and natural killer (NK) cells. The endoplasmic reticulum-resident human cytomegalovirus (HCMV) glycoprotein unique short (US) 6 and US11 genes prevent loading of MHC class antigen presentation to CD8+ T cells. Expression of Kaposi's sarcoma-associated herpesvirus (KSHV) K3 and K5 genes also inhibit the cytotoxic activity of CTL cells. Furthermore, K5 gene can inhibit the NK activating receptors. As a consequence, expression of these genes in porcine cells would inhibit CTL and NK cell mediated cytotoxicity in xenotransplantation. In this study, we transfected the US6, US11, K3 and K5 genes into minipig's fetal fibroblasts and established two transgenic clonal cell lines. The integration of these genes into minipig's fetal fibroblasts was confirmed by PCR and Southern blot. The reduction of Swine Leukocyte Antigen (SLA)-I by these genes was also detected by FACS assay. The level (46% to 55%) of SLA-I expression in complex cell lines was decreased relative to the control. In CTL assay, the rate of CD8+ T cell-mediated cytotoxicity was significantly reduced to 8.0 ± 7.0% comparing to the control (54.0 ± 6.0%, p<.05). In NK cytotoxicity assay, the rate of NK-92MI mediated cytotoxicity was significantly reduced to 43.4 ± 11.5% comparing to the control (76.7 ± 14.1%, p<.05). In conclusion, these results indicate that the expression of US6, US11, K3, K5 genes on porcine cells can effectively control CTL and NK cell mediated cytolysis. This strategy would provide a new approach to overcome CTL and NK-mediated rejection in xenotransplantation. |
| Databáze: |
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