TY - JOUR
T1 - Transfection of human pancreatic islets with an anti-apoptotic gene (bcl-2) protects β-cells from cytokine-induced destruction
AU - Rabinovitch, Alex
AU - Suarez-Pinzon, Wilma
AU - Strynadka, Ken
AU - Ju, Qida
AU - Edelstein, Diane
AU - Brownlee, Michael
AU - Korbutt, Gregory S.
AU - Rajotte, Ray V.
PY - 2000/6
Y1 - 2000/6
N2 - Apoptosis has been identified as a mechanism of pancreatic islet β- cell death in autoimmune diabetes. Proinflammatory cytokines are candidate mediators of β-cell death in autoimmune diabetes, and these cytokines can induce β-cell death by apoptosis. In the present study, we examined whether transfection of human islet β-cells with an anti-apoptotic gene, bcl-2, can prevent cytokine-induced β-cell destruction. Human islet β-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 β-galactosidase reporter gene (HSVlac). Two- color immunohistochemical staining revealed that 95 ± 3% of β-cells transfected with HSVbcl-2 expressed Bcl-2 protein compared with 14 ± 3% of β-cells transfected with HSVlac and 19 ± 4% of nontransfected β-cells. The bcl-2-transfected β-cells were fully protected from impaired insulin secretion and destruction resulting from incubation for 5 days with the cytokine combination of interleukin (IL)-1β, tumor necrosis factor (TNF)- α, and interferon (IFN)-γ. 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 β-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 β-cells in pancreatic islets, or islet grafts, from immune- mediated damage in type 1 diabetes.
AB - Apoptosis has been identified as a mechanism of pancreatic islet β- cell death in autoimmune diabetes. Proinflammatory cytokines are candidate mediators of β-cell death in autoimmune diabetes, and these cytokines can induce β-cell death by apoptosis. In the present study, we examined whether transfection of human islet β-cells with an anti-apoptotic gene, bcl-2, can prevent cytokine-induced β-cell destruction. Human islet β-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 β-galactosidase reporter gene (HSVlac). Two- color immunohistochemical staining revealed that 95 ± 3% of β-cells transfected with HSVbcl-2 expressed Bcl-2 protein compared with 14 ± 3% of β-cells transfected with HSVlac and 19 ± 4% of nontransfected β-cells. The bcl-2-transfected β-cells were fully protected from impaired insulin secretion and destruction resulting from incubation for 5 days with the cytokine combination of interleukin (IL)-1β, tumor necrosis factor (TNF)- α, and interferon (IFN)-γ. 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 β-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 β-cells in pancreatic islets, or islet grafts, from immune- mediated damage in type 1 diabetes.
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U2 - 10.2337/diabetes.48.6.1223
DO - 10.2337/diabetes.48.6.1223
M3 - Article
C2 - 10342808
AN - SCOPUS:0345161818
SN - 0012-1797
VL - 48
SP - 1223
EP - 1229
JO - Diabetes
JF - Diabetes
IS - 6
ER -