Contamination with E1A-positive wild-type adenovirus accounts for species-specific stimulation of islet cell proliferation by CCK: a cautionary note.
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.
Lavine
Jeremy A
JA
Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin, USA.
Raess
Philipp W
PW
Davis
Dawn B
DB
Rabaglia
Mary E
ME
Presley
Brent K
BK
Keller
Mark P
MP
Beinfeld
Margery C
MC
Kopin
Alan S
AS
Newgard
Christopher B
CB
Attie
Alan D
AD
D000256:Adenoviridae / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D017500:Adenovirus E1A Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001483:Base Sequence; D049109:Cell Proliferation; D002766:Cholecystokinin / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors; D006801:Humans; D007515:Islets of Langerhans / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D013045:Species Specificity; D014162:Transfection
D017500:Adenovirus E1A Proteins; D011498:Protein Precursors; D012333:RNA, Messenger; D002766:Cholecystokinin