Adenovirus-mediated gene expression in isolated rat pancreatic acini and individual pancreatic acinar cells.
In this study we have examined the feasibility of using replication-deficient recombinant adenoviral vectors to transfer and express genes in pancreatic acinar cells in vitro. We infected primary cultures of both isolated pancreatic acini and individual acinar cells with a recombinant adenovirus containing the coding sequence for beta-galactosidase. Our data demonstrate that recombinant adenoviruses readily infect pancreatic acinar cells in vitro. Close to 100% infection and maximal beta-galactosidase expression were obtained, when acini or acinar cells were infected with 5x10(6) or 10(6) plaque-forming units (pfu) of virus per millitre of acini or acinar cell suspension, respectively. Examination of the time-course of beta-galactosidase expression showed that there was a lag of approximately 6 h before beta-galactosidase levels increased. Thereafter beta-galactosidase expression increased rapidly. By 20 h post-infection beta-galactosidase activity had increased from undetectable levels to 2.5-3.0 units/mg of cellular protein. Acini/acinar cells maintained a robust secretory response after adenoviral infection. The cholecystokinin-octapeptide (CCK8) dose/response curves for amylase secretion for acini and acinar cells infected with 5x10(5) and 1x10(5) pfu/ml of virus, respectively, were biphasic, with maximal amylase secretion being stimulated by 1 nM CCK8. In addition, the dose/response curves were identical to those obtained from control, sham-infected, acini/acinar cells. Our findings indicate that replication-deficient recombinant adenoviral vectors will be excellent tools to transfer and express genes in isolated pancreatic acini or acinar cells.
Department of Pathology, Saint Louis University Health Sciences Center, St. Louis, MO, USA. padfield@wpogate.slu.edu
P J
PJ
Padfield
A C
AC
Elliott
J J
JJ
Baldassare
D000256:Adenoviridae / Q000235:genetics*; D000681:Amylases / Q000032:analysis / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D002766:Cholecystokinin / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005089:Exocytosis; D015870:Gene Expression / Q000235:genetics*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D008954:Models, Biological; D010179:Pancreas / Q000166:cytology* / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection / Q000379:methods; D019076:Transgenes / Q000235:genetics; D001616:beta-Galactosidase / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics
D011994:Recombinant Proteins; D002766:Cholecystokinin; D000681:Amylases; D001616:beta-Galactosidase