Adenovirus-Mediated Expression of the Anticoagulant Hirudin in Human Islets: A Tool to Make the Islets Biocompatible to Blood

Cell Transplantation

Article

Sanja Cabric¹  Olle Korsgren¹  Bo Nilsson¹  Graciela Elgue¹  Peter Schmidt¹ 

[1] Division of Clinical Immunology, The Rudbeck Laboratory, Uppsala University, Uppsala, Sweden;

关键词: Transplantation;    Coagulation;    Human islets of Langerhans;    Hirudin;    Adenoviral vector;   

DOI  :  10.3727/000000006783464390

 received in 2006-02-23, accepted in 2006-05-29,  发布年份 2006

来源: Sage Journals

【 摘 要 】

Human islets induce an injurious clotting reaction at the time of transplantation. A potential strategy to counteract this reaction would be to allow the islets to express hirudin, a protein with direct anticoagulative activity. Human islets were transduced with an adenoviral vector encoding hirudin, an empty corresponding vector, or left untreated. Islet culture supernatants were analyzed for hirudin using an ELISA, a chromogenic substrate assay based on the thrombin-binding properties of hirudin and in a whole blood viscosimetry assay. Immunohistochemical evaluation and determination of hirudin content revealed an abundant expression of hirudin after transduction. Hirudin content in transduced islets was in the range of the insulin content levels. A delay in human whole blood clotting time could be observed after addition of supernatants taken from islet cultures expressing hirudin. However, transduced islets showed an impaired glucose-stimulated insulin release, but could readily be retrieved 6 weeks after transplantation to athymic mice. A marked expression and secretion of hirudin with functional capacity can be induced in human islets using an adenoviral vector. The impairment in glucose-stimulated insulin release in hirudin-secreting islets, compared to controls, indicates that the additional protein synthesis affects the functional capacity of the islets.

【 授权许可】

Unknown   
© 2006 Cognizant Comm. Corp.