Adenoviral-mediated transduction of human pancreatic islets: importance of adenoviral genome for cell viability and association with a deficient antiviral response.
As adenoviral vectors are extensively used for genetic manipulation of insulin-producing cells in vitro, there is an increasing need to evaluate their effects on the function, morphology, and viability of transduced pancreatic islets. In the present study we observed that specific adenoviral genotypes, carrying E4 and E1/E3 deletions, correlate with differential induction of necrosis in pancreatic islet cells. In particular, the adenovirus death protein encoded from the E3 region of the adenoviral genome was able to modulate the changes induced in the morphology and viability of the transduced cells. We also propose a putative role for the transcriptional regulator pIX. Although human islet cells showed an increased resistance in terms of viral concentrations required for the induction of cell toxicity, our results showed that they were unable to build up an efficient antiviral response after transduction and that their survival was dependent on the exogenous addition of alpha-interferon. An intact and fully functional beta-cell is crucial for the successful application of gene therapy approaches in type 1 diabetes, and therefore, the implications of our findings need to be considered when designing vectors for gene transfer into pancreatic beta-cells.
Barbu
Andreea R
AR
Department of Medical Cell Biology, Uppsala University, Uppsala, Sweden. andreea.barbu@medcellbiol.uu.se
Akusjärvi
Göran
G
Welsh
Nils
N
D000244:Adenosine Diphosphate / Q000494:pharmacology; D000256:Adenoviridae / Q000235:genetics*; D017498:Adenovirus E1 Proteins / Q000235:genetics; D017503:Adenovirus E3 Proteins / Q000235:genetics; D017504:Adenovirus E4 Proteins / Q000235:genetics; D000818:Animals; D000998:Antiviral Agents / Q000032:analysis; D002470:Cell Survival; D002478:Cells, Cultured; D017353:Gene Deletion; D005822:Genetic Vectors; D006801:Humans; D016898:Interferon-alpha / Q000008:administration & dosage / Q000235:genetics; D007515:Islets of Langerhans / Q000737:chemistry / Q000473:pathology / Q000821:virology*; D009336:Necrosis; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D017498:Adenovirus E1 Proteins; D017503:Adenovirus E3 Proteins; D017504:Adenovirus E4 Proteins; D000998:Antiviral Agents; D016898:Interferon-alpha; D012333:RNA, Messenger; D000244:Adenosine Diphosphate