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-de ficient recombinant adenoviral vectors to transfer and express genes i n pancreatic acinar cells in vitro. We infected primary cultures of bo th isolated pancreatic acini and individual acinar cells with a recomb inant adenovirus containing the coding sequence for beta-galactosidase . Our data demonstrate that recombinant adenoviruses readily infect pa ncreatic acinar cells in vitro. Close to 100% infection and maximal be ta-galactosidase expression were obtained, when acini or acinar cells were infected with 5x10(6) or 10(6) plaque-forming units (pfu) of viru s per millitre of acini or acinar cell suspension, respectively. Exami nation 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 fro m undetectable levels to 2.5-3.0 units/mg of cellular protein. Acini/a cinar cells maintained a robust secretory response after adenoviral in fection. The cholecystokinin-octapeptide (CCK8) dose/response curves f or 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 dos e/response curves were identical to those obtained from control, sham- infected, acini/acinar cells. Our findings indicate that replication-d eficient recombinant adenoviral vectors will be excellent tools to tra nsfer and express genes in isolated pancreatic acini or acinar cells.