Monoclonal Antibodies to Intermediate Filament Proteins of Human Cells: Unique and Cross-reacting Antibodies

Overview

Journal J Cell Biol

Specialty Cell Biology

Date 1982 Nov 1

PMID 6183272

Citations 58

Authors

A M Gown

A M Vogel

Affiliations

Soon will be listed here.

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 indentification on SDS polyacrylamide gels ("wester blot" experiments), and immunoperoxidase assays on intact tissues. The following four antibodies are described: (a) an antivimentin antibody generated against human fibroblast cytoskeleton; (b), (c) two antibodies that recognize a 54-kdalton protein in human hepatocellular carcinoma cells; and (d) an antikeratin antibody made to stratum corneum that recognizes proteins of molecular weight 66 kdaltons and 57 kdaltons. The antivimentin antibody reacts with vimentin (58 kdaltons), glial fibrillary acidic protein (GFAP), 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. 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 molecular weights 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 antistratum 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.

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References

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

Dahl D, Bignami A . Immunochemical and immunofluorescence studies of the glial fibrillary acidic protein in vertebrates. Brain Res. 1973; 61:279-93. DOI: 10.1016/0006-8993(73)90533-7. View

Starger J, Goldman R . Isolation and preliminary characterization of 10-nm filaments from baby hamster kidney (BHK-21) cells. Proc Natl Acad Sci U S A. 1977; 74(6):2422-6. PMC: 432184. DOI: 10.1073/pnas.74.6.2422. View

Schachner M, Hedley-Whyte E, Hsu D, Schoonmaker G, Bignami A . Ultrastructural localization of glial fibrillary acidic protein in mouse cerebellum by immunoperoxidase labeling. J Cell Biol. 1977; 75(1):67-73. PMC: 2111556. DOI: 10.1083/jcb.75.1.67. View

Hynes R, Destree A . 10 nm filaments in normal and transformed cells. Cell. 1978; 13(1):151-63. DOI: 10.1016/0092-8674(78)90146-0. View

Sun T, Green H . Keratin filaments of cultured human epidermal cells. Formation of intermolecular disulfide bonds during terminal differentiation. J Biol Chem. 1978; 253(6):2053-60. View

Goldman J, Schaumburg H, Norton W . Isolation and characterization of glial filaments from human brain. J Cell Biol. 1978; 78(2):426-40. PMC: 2110133. DOI: 10.1083/jcb.78.2.426. View

Franke W, Weber K, Osborn M, Schmid E, Freudenstein C . Antibody to prekeratin. Decoration of tonofilament like arrays in various cells of epithelial character. Exp Cell Res. 1978; 116(2):429-45. DOI: 10.1016/0014-4827(78)90466-4. View

Franke W, Schmid E, Weber K, Osborn M . HeLa cells contain intermediate-sized filaments of the prekeratin type. Exp Cell Res. 1979; 118(1):95-109. DOI: 10.1016/0014-4827(79)90587-1. View

10.

Liem R, Yen S, Salomon G, Shelanski M . Intermediate filaments in nervous tissues. J Cell Biol. 1978; 79(3):637-45. PMC: 2110269. DOI: 10.1083/jcb.79.3.637. View

11.

Nowinski R, Lostrom M, Tam M, Stone M, Burnette W . The isolation of hybrid cell lines producing monoclonal antibodies against the p15(E) protein of ecotropic murine leukemia viruses. Virology. 1979; 93(1):111-26. DOI: 10.1016/0042-6822(79)90280-0. View

12.

Tuszynski G, FRANK E, Damsky C, Buck C, Warren L . The detection of smooth muscle desmin-like protein in BHK21/C13 fibroblasts. J Biol Chem. 1979; 254(13):6138-43. View

13.

Sun T, Shih C, Green H . Keratin cytoskeletons in epithelial cells of internal organs. Proc Natl Acad Sci U S A. 1979; 76(6):2813-7. PMC: 383699. DOI: 10.1073/pnas.76.6.2813. View

14.

Gard D, Bell P, Lazarides E . Coexistence of desmin and the fibroblastic intermediate filament subunit in muscle and nonmuscle cells: identification and comparative peptide analysis. Proc Natl Acad Sci U S A. 1979; 76(8):3894-8. PMC: 383942. DOI: 10.1073/pnas.76.8.3894. View

15.

Towbin H, Staehelin T, Gordon J . Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979; 76(9):4350-4. PMC: 411572. DOI: 10.1073/pnas.76.9.4350. View

16.

Schlegel R, Pinkus G . Immunohistochemical localization of keratin in normal human tissues. Lab Invest. 1980; 42(1):91-6. View

17.

Steinert P, Idler W, Goldman R . Intermediate filaments of baby hamster kidney (BHK-21) cells and bovine epidermal keratinocytes have similar ultrastructures and subunit domain structures. Proc Natl Acad Sci U S A. 1980; 77(8):4534-8. PMC: 349878. DOI: 10.1073/pnas.77.8.4534. View

18.

Franke W, Denk H, Kalt R, Schmid E . Biochemical and immunological identification of cytokeratin proteins present in hepatocytes of mammalian liver tissue. Exp Cell Res. 1981; 131(2):299-318. DOI: 10.1016/0014-4827(81)90234-2. View

19.

Yen S, Fields K . Antibodies to neurofilament, glial filament, and fibroblast intermediate filament proteins bind to different cell types of the nervous system. J Cell Biol. 1981; 88(1):115-26. PMC: 2111704. DOI: 10.1083/jcb.88.1.115. View

20.

Nelson W, Traub P . Properties of Ca2+-activated protease specific for the intermediate-sized filament protein vimentin in Ehrlich-ascites-tumour cells. Eur J Biochem. 1981; 116(1):51-7. DOI: 10.1111/j.1432-1033.1981.tb05299.x. View