Efficient ex vivo transduction of pancreatic islet cells with recombinant adeno-associated virus vectors

Terence Flotte, Anupam Agarwal, Jianming Wang, Sihong Song, Elizabeth S. Fenjves, Luca Inverardi, Kye Chesnut, Sandra Afione, Scott Loiler, Clive Wasserfall, Matthias Kapturczak, Tamir Ellis, Harry Nick, Mark Atkinson

Research output: Contribution to journalArticle

75 Scopus citations

Abstract

The ability to transfer immunoregulatory, cytoprotective, or antiapoptotic genes into pancreatic islet cells may allow enhanced posttransplantation survival of islet allografts and inhibition of recurrent autoimmune destruction of these cells in type 1 diabetes. However, transient transgene expression and the tendency to induce host inflammatory responses have limited previous gene delivery studies using viral transfer vectors. We demonstrate here that recombinant adeno-associated virus (rAAV) serotype 2, a vector that can overcome these limitations, effectively transduces both human and murine pancreatic islet cells with reporter genes as well as potentially important immunoregulatory cytokine genes (interleukin-4, interleukin-10), although a very high multiplicity of infection (10,000 infectious units/islet equivalent) was required. This requirement was alleviated by switching to rAAV serotype 5, which efficiently transduced islets at a multiplicity of infection of 100. Although adenovirus (Ad) coinfection was required for efficient ex vivo expression at early time points, islets transduced without Ad expressed efficiently when they were transplanted under the renal capsule and allowed to survive in vivo. The rAAV delivered transgenes did not interfere with islet cell insulin production and were expressed in both β- and non-β-cells. We believe rAAV will provide a useful tool to deliver therapeutic genes for modulating immune responses against islet cells and markedly enhance long-term graft survival.

Original languageEnglish (US)
Pages (from-to)515-520
Number of pages6
JournalDiabetes
Volume50
Issue number3
DOIs
StatePublished - Jan 1 2001

ASJC Scopus subject areas

  • Internal Medicine
  • Endocrinology, Diabetes and Metabolism

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    Flotte, T., Agarwal, A., Wang, J., Song, S., Fenjves, E. S., Inverardi, L., Chesnut, K., Afione, S., Loiler, S., Wasserfall, C., Kapturczak, M., Ellis, T., Nick, H., & Atkinson, M. (2001). Efficient ex vivo transduction of pancreatic islet cells with recombinant adeno-associated virus vectors. Diabetes, 50(3), 515-520. https://doi.org/10.2337/diabetes.50.3.515