Production of human monoclonal anti-basement membrane zone (BMZ) antibodies from a patient with bullous pemphigoid (BP) by Epstein-Barr virus transformation. Analyses of the heterogeneity of anti-BMZ antibodies in BP sera using them.
We established three lymphoblastoid cell lines from a bullous pemphigoid (BP) patient's peripheral blood by means of EBV transformation, which produced human monoclonal anti-basement membrane zone (BMZ) IgG antibodies. A blocking immunofluorescence test using these MAbs, designated 5A, 5E, and 10D, revealed that 5A and 5E recognized the same or a closely associated epitope, but the epitope for 10D was completely different, 18 of 30 BP sera blocked the reactivity of 10D MAb and 17 sera blocked 5E, while 9 sera did not block the staining of either antibody. Immunoblot analysis demonstrated that both 5A and 5E MAbs reacted exclusively with a protein band of approximately 230 kD in normal human epidermal extracts. However, 10D did not show any protein band. 22 of 30 BP sera strongly reacted with the same 230-kD protein, while none of control sera showed such reactivity. These results clearly demonstrated the heterogeneity of anti-BMZ antibodies in terms of epitopes. These MAbs should be useful in future investigations concerning not only the immunopathology but also the biochemial and molecular analyses of the BP antigen.
Sugi
T
T
Department of Dermatology, Keio University School of Medicine, Tokyo, Japan.
Hashimoto
T
T
Hibi
T
T
Nishikawa
T
T
D000368:Aged; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000916:Antibody Diversity; D001485:Basement Membrane / Q000276:immunology*; D001710:Biotin; D015153:Blotting, Western; D002472:Cell Transformation, Viral; D005455:Fluorescent Antibody Technique; D004854:Herpesvirus 4, Human; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology* / Q000302:isolation & purification; D008214:Lymphocytes / Q000382:microbiology; D008297:Male; D010391:Pemphigoid, Bullous / Q000276:immunology*; D012872:Skin Diseases, Vesiculobullous / Q000276:immunology*
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D001710:Biotin