title: Correction of the cystic fibrosis defect by gene complementation in human intrahepatic biliary epithelial cell lines.
abstract: OBJECTIVE
  Hepatobiliary disease is the second most common cause of mortality in patients with cystic fibrosis (CF). In the liver, only the intrahepatic biliary epithelial (IBE) cells express cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel. The aim of this study was to determine whether human CF-derived IBE cells can be infected with adenovirus and the CF phenotype complemented.
  
  
  METHODS
  IBE cells were isolated from 2 patients with CF and immortalized using retrovirus transduction of SV40 large T antigen. Immortalized cells were infected with the adenovirus vector Ad2/CFTR2 and assayed 2-31 days postinfection for cyclic adenosine monophosphate (cAMP)-induced halide efflux. Halide efflux was measured in single cells using fluorescence microscopy and the fluorescent probe 6-methoxy-N-(3-sulfopropyl)-quinolinium.
  
  
  RESULTS
  CF-derived IBE cell lines express biliary specific markers and express no cAMP-inducible halide efflux. Following infection with the adenovirus vector Ad2/CFTR2, a cAMP-induced halide efflux was observed for 31 days, although the number of responsive cells decreased with time.
  
  
  CONCLUSIONS
  Human CF-IBE cells can be infected by adenovirus and the defective CFTR complemented. The loss of responsive cells with time could be due to loss of construct and/or a reduced growth of cells that are overexpressing CFTR. These CF-IBE cell lines offer an opportunity to determine the mechanisms responsible for hepatobiliary disease in the patients with CF.
authors.affiliation: Department of Pediatrics, New England Medical Center, Boston, Massachusetts.
authors.forename: S A
authors.initials: SA
authors.lastname: Grubman
authors.forename: S L
authors.initials: SL
authors.lastname: Fang
authors.forename: A E
authors.initials: AE
authors.lastname: Mulberg
authors.forename: R D
authors.initials: RD
authors.lastname: Perrone
authors.forename: L C
authors.initials: LC
authors.lastname: Rogers
authors.forename: D W
authors.initials: DW
authors.lastname: Lee
authors.forename: D
authors.initials: D
authors.lastname: Armentano
authors.forename: S L
authors.initials: SL
authors.lastname: Murray
authors.forename: H L
authors.initials: HL
authors.lastname: Dorkin
authors.forename: S H
authors.initials: SH
authors.lastname: Cheng
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001652:Bile Ducts / Q000378:metabolism*; D002460:Cell Line; D003550:Cystic Fibrosis / Q000628:therapy*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004848:Epithelium; D005816:Genetic Complementation Test; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics*; D051379:Mice; D014162:Transfection
chemical_list: C505032:CFTR protein, human; D008565:Membrane Proteins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator