title: Characterization of a SV40-transformed rheumatoid synovial fibroblast cell line which retains genotypic expression patterns: a model for evaluation of anti-arthritic agents.
abstract: A chimeric Adenovirus-Simian Virus 40 (AdSV40) containing the large T antigen was used to transform rheumatoid synovial fibroblasts. A rheumatoid synovial fibroblast cell line was established by infection of primary rheumatoid arthritis (RA) synovial fibroblasts at Passage 10 with AdSV40 recombinants followed by selection in semisoft agarose cultures. The transformed cells grew anchor independent, exhibited continuous proliferation (> 65 passages) in monolayer culture, and formed multiple visible foci. The transformed synovial fibroblasts showed expression of the simian virus 40 large T antigen in the nucleus as determined by immunofluorescence staining. In addition, indirect immunofluorescence staining demonstrated that the transformed cells stained specifically with a fibroblast-specific antibody 1B10. Studies involving expression of metalloproteinases showed that collagenase and stromelysin were induced by phorbal 12-myristate 13-acetate (PMA), and such an induction was repressed by dexamethasone typical of primary RA fibroblasts. Levels of mRNAs for IL-1 beta, TNF-alpha, and c-jun were increased by PMA, and the mRNA transcripts of these genes were also repressed by addition of dexamethasone to the culture media. Our results indicate that transformed RA synovial fibroblasts display a similar gene expression pattern in response to PMA and dexamethasone as observed for untransformed primary RA synovial fibroblasts. These transformed rheumatoid arthritis synovial fibroblast cells provide an ideal cell culture model in which to test the efficacy of novel arthritis gene therapy reagents.
authors.affiliation: University of Alabama at Birmingham, Department of Medicine 35294, USA.
authors.forename: H G
authors.initials: HG
authors.lastname: Zhang
authors.forename: W D
authors.initials: WD
authors.lastname: Blackburn
authors.forename: P P
authors.initials: PP
authors.lastname: Minghetti
mesh_terms: D018501:Antirheumatic Agents / Q000627:therapeutic use*; D001172:Arthritis, Rheumatoid / Q000188:drug therapy / Q000201:enzymology / Q000473:pathology*; D002461:Cell Line, Transformed; D003907:Dexamethasone / Q000494:pharmacology; D005347:Fibroblasts / Q000201:enzymology / Q000473:pathology; D005455:Fluorescent Antibody Technique; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D016758:Genes, jun; D005838:Genotype; D006801:Humans; D007375:Interleukin-1 / Q000235:genetics; D008666:Metalloendopeptidases / Q000235:genetics; D008954:Models, Biological; D013539:Simian virus 40 / Q000502:physiology*; D013583:Synovial Membrane / Q000201:enzymology / Q000473:pathology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D016211:Transforming Growth Factor alpha / Q000235:genetics
chemical_list: D018501:Antirheumatic Agents; D007375:Interleukin-1; D016211:Transforming Growth Factor alpha; D003907:Dexamethasone; D008666:Metalloendopeptidases; D013755:Tetradecanoylphorbol Acetate