Anti-Fas antibody induces apoptosis in cultured human renal interstitial fibroblasts
- VernacularTitle:抗Fas抗体诱导人肾间质成纤维细胞凋亡
- Author:
Lijing CHENG
1
;
Yipu CHEN
Author Information
1. 北京医科大学附属第一医院
- Keywords:
apoptosis;
interstitial fibroblasts;
Fas;
anti-Fas antibody
- From:
Chinese Medical Journal
2001;114(2):162-166
- CountryChina
- Language:Chinese
-
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 (γ-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 γ-IFN (500 U/ml, 48 hours) were incubated with anti-Fas antibody (IgM, 0.5 μg/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 γ-IFN. Apoptosis in γ-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. Conclusion Fas is normally expressed in hRIFs and can be markedly upregulated by γ-IFN. Anti-Fas antibodies can induce apoptosis of hRIFs pre-stimulated with γ-IFN.
