Contamination with E1A-positive wild-type adenovirus accounts for species-specific stimulation of islet cell proliferation by CCK: a cautionary note

Mol Endocrinol. 2010 Feb;24(2):464-7. doi: 10.1210/me.2009-0384. Epub 2010 Jan 15.

Abstract

We have previously reported that adenovirus-mediated expression of preprocholecystokin (CCK) stimulates human and mouse islet cell proliferation. In follow-up studies, we became concerned that the CCK adenovirus might have been contaminated with a wild-type E1A-containing adenovirus. Here we show conclusively that the proliferative effects reported in the original paper in mouse and human islets were not due to CCK expression but rather to a contaminating E1A-expressing wild-type adenovirus. We also show, however, that CCK expression does have a proliferative effect in rat islets. We hope that our report of the steps taken to detect the wild-type virus contamination, and purification of the contributing viral stocks, will be helpful to other investigators, and that our experience will serve as a cautionary tale for use of adenovirus vectors, especially for studies on cellular replication.

Publication types

  • Comment
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae / chemistry
  • Adenoviridae / genetics*
  • Adenoviridae / isolation & purification
  • Adenoviridae / metabolism
  • Adenovirus E1A Proteins / chemistry
  • Adenovirus E1A Proteins / genetics
  • Adenovirus E1A Proteins / metabolism*
  • Animals
  • Base Sequence
  • Cell Proliferation*
  • Cholecystokinin / genetics
  • Cholecystokinin / metabolism*
  • Genetic Vectors
  • Humans
  • Islets of Langerhans / cytology*
  • Islets of Langerhans / metabolism
  • Mice
  • Molecular Sequence Data
  • Protein Precursors / genetics
  • Protein Precursors / metabolism*
  • RNA, Messenger / chemistry
  • RNA, Messenger / metabolism
  • Rats
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Alignment
  • Species Specificity
  • Transfection*

Substances

  • Adenovirus E1A Proteins
  • Protein Precursors
  • RNA, Messenger
  • Cholecystokinin