國立臺灣大學醫學院外科李章銘2006-07-262018-07-112006-07-262018-07-112004-07-31http://ntur.lib.ntu.edu.tw//handle/246246/24521以豬器官來源的異種器官，為目前臨床移植所面臨的器官短缺的問題，提供了解決之道。 但是其可行性，目前仍受限於人類對豬隻的異種抗體所誘發的超急性，及亞急性異種器官排 斥，及物種間組織抗原歧異所引起的T 細胞排斥。先前在我們所產製的HLA DPW0401 基因轉殖 豬中，發現人類HLA DP 的抗原性，可完整表達在豬細胞表面，而相較於其他非基因轉殖同胞 豬，HLA DPW0401 基因轉殖豬所引起人類周邊單核球(具HLA DPW0401 基因型者)的增生反應， 有減少的趨勢。 HO-1 是代謝Heme 的主要酵素，而能保護細胞抵抗異種抗體所引發的細胞裂解及凋亡,及 使異種移植器官在異種抗體存在之下，達到”適存化”的狀態。而人類延遲加速因子(hDAF, CD55)為一補體活化鏈的調節者。目前發現 hDAF 的基因轉殖豬器官，包括心臟，腎臟及肝臟 可在靈長類體內， 存活7 天至3 個月。 在過去一年中，我們已經成功地產製了HLA-DR 及HO-1 之基因轉殖豬，其mRNA 及其蛋白 的表達都已得到確定，針對HO-1 保護作用的測試，我們已經成功地完成自然殺手細胞毒性測 試的檢驗模式，我們發現殺手細胞與標的細胞於5：1 的情形有最明顯的毒殺現象，之後我們 將運用於HO-1 轉殖豬細胞的測試，檢測此HO-1 轉殖基因是否有保護作用。我們也完成了 HLA-DR 基因轉殖豬細胞對人類週邊單核球的增生測試，我們發覺這種基因轉殖豬細胞，與非 基因轉殖同胞豬細胞相較，其引起的異種細胞免疫反應，並沒有明顯的差別，至於這些轉殖豬 細胞，是否對於其他的異種免疫反應有保護作用，則有待進一步確定。Pig to human xenotransplatation is a promising strategy to overcome the organ shortage in clinical transplantation. However, the application is hampered by the xenoreactive antibody mediated hyperacute rejection, delayed xenograft rejection and MHC disparity associated T cell rejection. Previous, we have produced the HLA DPW0401 transgenic pig, which was shown equipped with the human HLA antigenecity and, as compared to the non-transgenic litermate pig, induced a minor cellular response to the human PBMCs that came from the HLA DPW 0401(+) human. HO-1 (heme oxygenase-1) is the main enzyme metabolizing the heme, which has been demonstrated to offer a protective effect for the xenograft to achieve accommodation under the presence of xenoreactive antibodies, and help to attenuate xenoantibody mediated celll lysis and apoptosis. To test the protective effect of HO-1 exogene from the NK related cytotoxicity, we have establish the optimal condition for this cytotoxic assay using the NK cell line, K562. The cell killing efficacy will be maximal when the effector/target ratio being 5: 1, 47% of cell death of the total cell population. We have also successfully cultured the aortic endothelial cells of pig (PAEC) from the non-transgenic pig. The culture of the PAEC of the transgenic pig is ongoing now. Because the HLA-DR 1501 has been successfully produced, we also test the xenogenic MLC for the HLA DR transgenic pig. We found that the tendency about xenogenic cellular response was not evidently different between the HLADR transgenic and non-transgenic littermate control pis PBMC. Whether this transgenic pig cell can obtain protective effect in another process of xenogenic immune response required to be examined further.application/pdf114435 bytesapplication/pdfzh-TW國立臺灣大學醫學院外科基因轉殖豬人類白血球抗原人類延遲加速因子第一型Heme 氧化酵素Transgenic PigXenotransplantationHLAHO-1hDAFreporthttp://ntur.lib.ntu.edu.tw/bitstream/246246/24521/1/922314B002226.pdf