Monoclonal antibodies to intermediate filament proteins of human cells: unique and cross- reacting antibodies

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Abstract
Monoclonal antibodies were generated against the intermediate filament proteins of different human cells. The reactivity of these antibodies with the different classes of intermediate filament proteins was determined by indirect immunofluorescence on cultured cells, immunologic identification on SDS[sodium dodecyl sulfate) polyacrylamide gels (western blot experiments), and immunoperoxidase assays on intact tissues. The following 4 antibodies are described: an antivimentin antibody generated against human fibroblast cytoskeleton; 2 antibodies that recognize a 54 kdalton protein in human hepatocellular carcinoma cells; and an antikeratin antibody made to stratum corneum that recognizes proteins of MW 66 kdaltons and 57 kdaltons. The antivimentin antibody reacts with vimentin (58 kdaltons), glial fibrillary acidic protein (GEAP) and keratins from stratum corneum, but does not recognize hepatoma intermediate filaments. In immunofluorescence assays, the antibody reacts with mesenchymal cells and cultured epithelial cells that express vimentin. This antibody decorates the media of blood vessels in tissue sections. One antihepatoma filament antibody reacts only with the 54 kdalton protein of these cells and, in immunofluorescence and immunoperoxidase assays, only recognizes epithelial cells. It reacts with almost all nonsquamous epithelium in intact tissue, but fails to recognize squamous epithelium. Thus this antibody identifies an epithelial-specific cytokeratin molecule present in only nonsquamous epithelium. The other antihepatoma filament antibody is much less selective, reacting with vimentin, GFAP and keratin from stratum corneum. This antibody decorates intermediate filaments of both mesenchymal and epithelial cells. The antikeratin antibody recognizes 66 kdalton and 57 kdalton proteins in extracts of stratum corneum and also identifies proteins of similar MW in all cells tested. However, by immunofluorescence, this antibody decorates only the intermediate filaments of epidermoid carcinoma cells. When assayed on tissue sections, the antibody reacts with squamous epithelium and some, but not all, nonsquamous epithelium. Therefore this anitstratum corneum antibody and the anti-54 kdalton antibody identify unique epitopes present in the various cytokeratin molecules of epithelial cells. None of the hybridoma antibodies react with neurofilament proteins. The different patterns of reactivity of these antibodies suggest that many of the immunologically distinct intermediate filament proteins contain common antigenic determinants.