title: Anti-Fas antibody induces apoptosis in cultured human renal interstitial fibroblasts.
abstract: OBJECTIVE
  To detect if Fas is expressed in human renal interstitial fibroblasts (hRIFs) and apoptosis of hRIFs can be induced by specific anti-Fas antibody.
  
  
  METHODS
  hRIFs were cultured from isolated papillae of human kidney, and identified by morphologic examination, assay of antigenic components and culture in D-valine selective medium. Fas expression in normal hRIFs was detected by RT-PCR and immunocytochemistry staining. After hRIFs were incubated with interferon gamma (gamma-IFN 500 U/ml, 1000 U/ml, 1500 U/ml and 2000 U/ml, respectively) for 48 hours, Fas expression was determined by Northern blot, Western blot and flow cytometry. hRIFs pre-stimulated with gamma-IFN (500 U/ml, 48 hours) were incubated with anti-Fas antibody (IgM, 0.5 microgram/ml) for 12 hours. And apoptosis was identified by morphologic examination, DNA ladder assay and flow cytometry.
  
  
  RESULTS
  The cultured hRIFs showed a shuttle-like shape and were positively stained by labeled anti-vimentin antibody but negatively stained by anti-epithelial membrane antibody. They could not grow in the D-valine selective medium and died partly in a week. Fas mRNA and protein were expressed in normal hRIFs and markedly upregulated by stimulation with gamma-IFN. Apoptosis in gamma-IFN pre-stimulated hRIFs was induced by anti-Fas antibody, showing cell nuclear shrinkage and condensation in morphologic feature, internucleosomal DNA fragmentation in DNA ladder assay and a pick of hypo-diploid nuclei by flow cytometry.
  
  
  CONCLUSIONS
  Fas is normally expressed in hRIFs and can be markedly upregulated by gamma-IFN. Anti-Fas antibodies can induce apoptosis of hRIFs pre-stimulated with gamma-IFN.
authors.affiliation: Division of Nephrology, the First Hospital, Beijing Medical University, Beijing 100034, China.
authors.forename: L
authors.initials: L
authors.lastname: Cheng
authors.forename: Y
authors.initials: Y
authors.lastname: Chen
mesh_terms: D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D007668:Kidney / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D019014:fas Receptor / Q000235:genetics / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D012333:RNA, Messenger; D019014:fas Receptor; D007371:Interferon-gamma