Gainer A L, Suarez-Pinzon W L, Min W P, Swiston J R, Hancock-Friesen C, Korbutt G S, Rajotte R V, Warnock G L, Elliott J F
Surgical-Medical Research Institute and Department of Surgery, University of Alberta, Edmonton, Canada.
Transplantation. 1998 Jul 27;66(2):194-9. doi: 10.1097/00007890-199807270-00009.
Pancreatic islet transplantation is limited because of immune rejection of the transplanted tissue. Long-term survival of allogeneic pancreatic islet grafts in the absence of systemic immunosuppressive agents should be possible by transfecting the islets directly with DNA encoding immunoregulatory molecules. Localized production of these molecules should affect only the immune cells that come into the vicinity of the foreign tissue. We investigated whether local expression of human CTLA4-Ig or soluble human Fas ligand from biolistically transfected mouse islets would have a protective effect on allograft survival.
Isolated CBA (H2k) islets were biolistically transfected using the gene gun. The experimental groups were naked gold particles (n=6), empty vector DNA (n=5), DNA encoding human CTLA4-Ig (n=8), or soluble human Fas ligand (n=5). Secretion of the transfected gene product was confirmed by screening islet culture supernatants for protein production using a sandwich ELISA. The blasted islets were transplanted under the kidney capsule of alloxan-diabetic BALB/c (H2d) recipients.
Control grafts survived for 23 days, on average. CTLA4-Ig-transfected islets showed a bimodal distribution: 50% of cases survived > or = 46 days and 50% were similar to the controls. In the soluble human Fas ligand group, 80% of grafts survived > or = 50 days. There was no correlation between graft survival times and pretransplant levels of protein production.
Our results indicate that local production of human CTLA4-Ig or soluble human Fas ligand by biolistically transfected islets can promote allograft survival. This approach should be valuable as a potential immunoprotective therapeutic strategy in tissue transplantation.
由于移植组织的免疫排斥反应，胰岛移植受到限制。通过用编码免疫调节分子的DNA直接转染胰岛，在不使用全身免疫抑制剂的情况下，异体胰岛移植的长期存活应该是可行的。这些分子的局部产生应该只影响进入外来组织附近的免疫细胞。我们研究了通过生物弹道转染的小鼠胰岛局部表达人CTLA4-Ig或可溶性人Fas配体是否会对同种异体移植存活产生保护作用。
使用基因枪对分离的CBA（H2k）胰岛进行生物弹道转染。实验组为裸金颗粒（n = 6）、空载体DNA（n = 5）、编码人CTLA4-Ig的DNA（n = 8）或可溶性人Fas配体（n = 5）。通过夹心ELISA筛选胰岛培养上清液中的蛋白质产生，以确认转染基因产物的分泌。将轰击后的胰岛移植到四氧嘧啶糖尿病BALB/c（H2d）受体的肾包膜下。
对照移植物平均存活23天。CTLA4-Ig转染的胰岛呈现双峰分布：50%的病例存活≥46天，50%与对照组相似。在可溶性人Fas配体组中，80%的移植物存活≥50天。移植物存活时间与移植前蛋白质产生水平之间没有相关性。
我们的结果表明，通过生物弹道转染的胰岛局部产生人CTLA4-Ig或可溶性人Fas配体可以促进同种异体移植存活。这种方法作为组织移植中一种潜在的免疫保护治疗策略应该是有价值的。
