Transduction of rat pancreatic islets with pseudotyped adeno-associated virus vectors.

Abstract

BACKGROUND:

Pancreatic islet transplantation is a promising treatment for type I diabetes mellitus, but current immunosuppressive strategies do not consistently provide long-term survival of transplanted islets. We are therefore investigating the use of adeno-associated viruses (AAVs) as gene therapy vectors to transduce rat islets with immunosuppressive genes prior to transplantation into diabetic mice.

RESULTS:

We compared the transduction efficiency of AAV2 vectors with an AAV2 capsid (AAV2/2) to AAV2 vectors pseudotyped with AAV5 (AAV2/5), AAV8 (AAV2/8) or bovine adeno-associated virus (BAAV) capsids, or an AAV2 capsid with an insertion of the low density lipoprotein receptor ligand from apolipoprotein E (AAV2apoE), on cultured islets, in the presence of helper adenovirus infection to speed expression of a GFP transgene. Confocal microscopy and flow cytometry were used. The AAV2/5 vector was superior to AAV2/2 and AAV2/8 in rat islets. Flow cytometry indicated AAV2/5-mediated gene expression in approximately 9% of rat islet cells and almost 12% of insulin-positive cells. The AAV2/8 vector had a higher dependence on the helper virus multiplicity of infection than the AAV 2/5 vector. In addition, the BAAV and AAV2apoE vectors were superior to AAV2/2 for transducing rat islets. Rat islets (300 per mouse) transduced with an AAV2/5 vector harboring the immunosuppressive transgene, tgf beta 1, retain the ability to correct hyperglycemia when transplanted into immune-deficient diabetic mice.

CONCLUSION:

AAV2/5 vectors may therefore be useful for pre-treating donor islets prior to transplantation.

• Authors: Anthony T Craig, Oksana Gavrilova, Nancy K Dwyer, William Jou, Stephanie Pack, Eric Liu, Klaus Pechhold, Michael Schmidt, Victor J McAlister, John A Chiorini, E Joan Blanchette-Mackie, David M Harlan, Roland A Owens
• Journal: Virology journal (Virol J) Vol. 6 Pg. 61 (May 18 2009) ISSN: 1743-422X [Electronic] England
• PMID: 19450275 (Publication Type: Evaluation Study, Journal Article, Research Support, N.I.H., Intramural)

Topics

• Animals
• Cell Line
• Dependovirus (genetics, metabolism)
• Diabetes Mellitus, Type 1 (genetics, surgery, therapy)
• Female
• Genetic Therapy (methods)
• Genetic Vectors (genetics, metabolism)
• Humans
• In Vitro Techniques
• Islets of Langerhans (metabolism, virology)
• Islets of Langerhans Transplantation
• Mice
• Mice, Inbred NOD
• Mice, SCID
• Rats
• Rats, Wistar
• Transduction, Genetic (methods)