Establish lentiviral vector system and apply to islet transplantation in autoimmune diabetes

• Main Authors: Yung-Liang Liu, 劉勇良
• Other Authors: Huey-Kang Sytwu
• Format: Others
• Language: en_US
• Published: 2004
• Online Access: http://ndltd.ncl.edu.tw/handle/04998604185143782754

碩士 === 國防醫學院 === 微生物及免疫學研究所 === 92 === Insulin-dependent diabetes mellitus (IDDM) is caused by a progressive autoimmune destruction of the insulin-producing βcells in the pancreatic islets of Langerhans. Islet transplantation in conjunction with glucocorticoid-free immunosuppressive regimen has proven to be a successful and curative approach to treating diabetes. However toxicity and possible infections accompanying the life-long recipient immunosuppression still limit the current application for islet transplantation. Transplantation of genetically engineered islets, a localized approach to regulating autoimmune responses to islet antigens in addition to allogeneic transplantation responses will be possible to be free of life-long immunosuppression. Based on the idea, we use an ideal gene transfer vector, lentivirus, which can transduce non-dividing cells, integrate into the cell genome and be non-immunogenic, to carry the genes with immunomodulatory potential, including surface-linked single chain antibody against CTLA-4 (scFv anti-CTLA-4), PD-L1 or PD-L2, and to transduce NIT-1 cells in various combinations in order to investigate the most potential regiemen and apply to islet transplantation. From in vitro proliferation assay, our preliminary data show that NIT-1 cells overexpressing surface-linked single chain antibody against CTLA-4 reveal inhibitory effects. Also in allograft experiments, islet cells transduced by lentiviral vectors carrying scFv anti-CTLA-4 in vitro for 2 days and transplanted to diabetic NOD mice, a widely used animal model for dissecting immunopathological mechanism in IDDM, show longer survival than the control group. In summary, we have established lentivirus and islet transplantation systems in this study. Cotransducing other transgenes to islets by lentiviral vectors to get better and longer survival is ongoing work in our lab.