Liver cell-membrane specific antibodies in autoimmune hepatitis patients detected by flow cytometry |
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| Affiliation: | 1. Division of Medicine, Institute of Gastroenterology, Tokyo Women''s Medical College, Tokyo, Japan;2. Department of Pathology, Juntendo University School of Medicine, Tokyo, Japan;1. Department of Medicine II (Gastroenterology, Hepatology, Endocrinology and Infectious Diseases), Freiburg University Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, Germany;2. Faculty of Biology, University of Freiburg, Freiburg, Germany;3. Institute of Medical Microbiology, Virology and Hygiene, University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany;4. German Center for Infection Research (DZIF), Hamburg-Lübeck-Borstel-Riems, Germany;5. Institute of Virology, Freiburg University Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, Germany;6. Department of Medicine, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany;7. Institute for Transfusion Medicine and Gene Therapy, Freiburg University Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, Germany;8. Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff, United Kingdom;9. Systems Immunity Research Institute, Cardiff University School of Medicine, Cardiff, United Kingdom;10. Department of Medicine IV (Nephrology and Primary Care), Freiburg University Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, Germany;11. Department of Rheumatology and Clinical Immunology, Freiburg University Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, Germany |
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| Abstract: | ![]()
In this study, we identified autoantibody in patients with autoimmune hepatitis against liver cell membrane using flow cytometry. After incubation of one of the hepatoblastoma cell lines, Hep G2, with serum from a patient, and the addition of FITC-labeled anti-human Ig antibody, anti-membrane antibodies were analyzed by flow cytometry. By our method, the antibody in serum can react only with autoantigens on the cell surface. Furthermore, propidium iodide staining enabled us to exclude the possibility of crossreaction of antibodies against dead cells. The relative fluorescence intensity in 12 patients with autoimmune chronic active hepatitis was significantly higher than that in 20 normal subjects, six primary biliary cirrhosis (PBC), 18 chronic viral hepatitis, and 11 systemic lupus erythematosus (SLE). The normal level of fluorescence intensity provided by sera from the SLE patients indicated that antibody binding to the liver cell membrane was not derived from Fc-mediated immune complex capture. These findings demonstrated that this flow cytometric technique provides a simple and accurate method for the detection of autoantibodies against liver cell membrane. |
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