Transfection of human pancreatic islets with an anti-apoptotic gene (bcl-2) protects β-cells from cytokine-induced destruction
Transfection of human pancreatic islets with an anti-apoptotic gene (bcl-2) protects β-cells from cytokine-induced destruction
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DOI：
10.2337/diabetes.48.6.1223
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发表时间：
1999-06-01
期刊：
影响因子：
7.7
通讯作者：
Rajotte, RV
中科院分区：
文献类型：
--
作者：
Rabinovitch, A;Suarez-Pinzon, W;Rajotte, RV
细胞凋亡已被确定为自身免疫性糖尿病中胰岛β细胞死亡的机制。促炎性细胞因子是自身免疫性糖尿病中β细胞死亡的候选介质，并且这些细胞因子可以通过凋亡诱导β细胞死亡。在本研究中，我们检测了用抗凋亡基因bcl-2转染人胰岛β细胞是否可以防止甘氨酸诱导的β细胞破坏。用表达bcl-2基因的复制缺陷型单纯疱疹病毒（HSV）扩增子载体（HSVbcl-2）转染人胰岛β细胞，并用表达β-半乳糖苷酶报告基因的相同HSV载体（HSVlac）作为对照。双色免疫组化染色显示，95 +/- 3%的转染HSV bcl-2的β细胞表达Bcl-2蛋白，而转染HSV Vlac的β细胞为14 +/- 3%，未转染的β细胞为19 +/- 4%。bcl-2转染的β细胞被完全保护免于受损的胰岛素分泌和与白细胞介素（IL）-1 β、肿瘤坏死因子（TNF）-α和干扰素（IFN）-γ的细胞因子组合孵育5天所导致的破坏。此外，bcl-2转染的胰岛细胞被显著保护免于奎宁诱导的脂质过氧化和DNA断裂。这些结果表明，甜菜碱诱导的β细胞功能障碍和死亡涉及受抗凋亡蛋白Bcl-2调节的机制。因此，bcl-2基因治疗具有保护胰岛或胰岛移植物中的人β细胞免受1型糖尿病中免疫介导的损伤的潜力。
Apoptosis has been identified as a mechanism of pancreatic islet beta-cell death in autoimmune diabetes. Proinflammatory cytokines are candidate mediators of beta-cell death in autoimmune diabetes, and these cytokines can induce beta-cell death by apoptosis, In the present study, we examined whether transfection of human islet beta-cells with an anti-apoptotic gene, bcl-2, can prevent cytokine-induced beta-cell destruction. Human islet beta-cells were transfected by a replication-defective herpes simplex virus (HSV) amplicon vector that expressed the bcl-2 gene (HSVbcl-2) and, as a control, the same HSV vector that expressed a beta-galactosidase reporter gene (HSVlac). Two-color immunohistochemical staining revealed that 95 +/- 3% of beta-cells transfected with HSVbcl-2 expressed Bcl-2 protein compared with 14 +/- 3% of beta-cells transfected with HSVlac and 19 +/- 4% of nontransfected beta-cells. The bcl-2-transfected beta-cells were fully protected from impaired insulin secretion and destruction resulting from incubation for 5 days with the cytokine combination of interleukin (IL)-1 beta, tumor necrosis factor (TNF)-alpha, and interferon (IFN)-gamma, In addition, the bcl-2-transfected islet cells were significantly protected from cytokine-induced lipid peroxidation and DNA fragmentation. These results demonstrate that cytokine-induced beta-cell dysfunction and death involve mechanisms subject to regulation by an anti-apoptotic protein, Bcl-2. Therefore, bcl-2 gene therapy has the potential to protect human beta-cells in pancreatic islets, or islet grafts, from immune-mediated damage in type 1 diabetes.