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. 1983;2(12):2305–2312. doi: 10.1002/j.1460-2075.1983.tb01738.x

Monoclonal antibodies to desmin, the muscle-specific intermediate filament protein.

E Debus, K Weber, M Osborn
PMCID: PMC555449  PMID: 6421570

Abstract

A set of monoclonal antibodies to desmin has been isolated from a fusion of mouse myeloma cells with spleen cells from mice immunized with purified porcine desmin. Eleven group I antibodies recognized desmin in the immune blot, and using defined desmin fragments the epitope has been tentatively assigned as lying between residues 325 and 372. When cell lines were tested in immunofluorescence only the human line RD and hamster BHK-21 were positive. When tissue sections were used, skeletal, cardiac, visceral and some vascular smooth muscle cells were positive. Thus, the group I antibodies appear specific for desmin and do not recognize other intermediate filament proteins. Group II monoclonals recognized not only desmin in the immune blot but also other polypeptides. The epitope of this class is located between residues 70 and 280. In immunofluorescence on cell lines and tissues, the staining patterns of group II antibodies were more complicated and demonstrate that not only other intermediate filament proteins but also additional antigenic determinants are being recognized. The group I antibodies stain, as expected from their desmin specificity, rat and human rhabdomyosarcomas and thus appear to be useful reagents in pathology.

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Selected References

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  1. Altmannsberger M., Osborn M., Hölscher A., Schauer A., Weber K. The distribution of keratin type intermediate filaments in human breast cancer. An immunohistological study. Virchows Arch B Cell Pathol Incl Mol Pathol. 1981;37(3):277–284. doi: 10.1007/BF02892576. [DOI] [PubMed] [Google Scholar]
  2. Altmannsberger M., Osborn M., Treuner J., Hölscher A., Weber K., Shauer A. Diagnosis of human childhood rhabdomyosarcoma of antibodies to desmin, the structural protein of muscle specific intermediate filaments. Virchows Arch B Cell Pathol Incl Mol Pathol. 1982;39(2):203–215. doi: 10.1007/BF02892848. [DOI] [PubMed] [Google Scholar]
  3. Anderton B. H., Breinburg D., Downes M. J., Green P. J., Tomlinson B. E., Ulrich J., Wood J. N., Kahn J. Monoclonal antibodies show that neurofibrillary tangles and neurofilaments share antigenic determinants. Nature. 1982 Jul 1;298(5869):84–86. doi: 10.1038/298084a0. [DOI] [PubMed] [Google Scholar]
  4. Bennett G. S., Fellini S. A., Toyama Y., Holtzer H. Redistribution of intermediate filament subunits during skeletal myogenesis and maturation in vitro. J Cell Biol. 1979 Aug;82(2):577–584. doi: 10.1083/jcb.82.2.577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Campbell G. R., Chamley-Campbell J., Gröschel-Stewart U., Small J. V., Anderson P. Antibody staining of 10-nm (100-A) filaments in cultured smooth, cardiac and skeletal muscle cells. J Cell Sci. 1979 Jun;37:303–322. doi: 10.1242/jcs.37.1.303. [DOI] [PubMed] [Google Scholar]
  6. Dahl D., Bignami A. Immunohistological localization of desmin, the muscle-type 100 A filament protein, in rat astrocytes and Müller glia. J Histochem Cytochem. 1982 Mar;30(3):207–213. doi: 10.1177/30.3.7037941. [DOI] [PubMed] [Google Scholar]
  7. Debus E., Weber K., Osborn M. Monoclonal cytokeratin antibodies that distinguish simple from stratified squamous epithelia: characterization on human tissues. EMBO J. 1982;1(12):1641–1647. doi: 10.1002/j.1460-2075.1982.tb01367.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Edström L., Thornell L. E., Eriksson A. A new type of hereditary distal myopathy with characteristic sarcoplasmic bodies and intermediate (skeletin) filaments. J Neurol Sci. 1980 Aug;47(2):171–190. doi: 10.1016/0022-510x(80)90002-7. [DOI] [PubMed] [Google Scholar]
  9. Fidzianska A., Goebel H. H., Osborn M., Lenard H. G., Osse G., Langenbeck U. Mallory body-like inclusions in a hereditary congenital neuromuscular disease. Muscle Nerve. 1983 Mar-Apr;6(3):195–200. doi: 10.1002/mus.880060305. [DOI] [PubMed] [Google Scholar]
  10. Frank E. D., Warren L. Aortic smooth muscle cells contain vimentin instead of desmin. Proc Natl Acad Sci U S A. 1981 May;78(5):3020–3024. doi: 10.1073/pnas.78.5.3020. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fuchs E., Green H. The expression of keratin genes in epidermis and cultured epidermal cells. Cell. 1978 Nov;15(3):887–897. doi: 10.1016/0092-8674(78)90273-8. [DOI] [PubMed] [Google Scholar]
  12. Gabbiani G., Kapanci Y., Barazzone P., Franke W. W. Immunochemical identification of intermediate-sized filaments in human neoplastic cells. A diagnostic aid for the surgical pathologist. Am J Pathol. 1981 Sep;104(3):206–216. [PMC free article] [PubMed] [Google Scholar]
  13. Geisler N., Kaufmann E., Fischer S., Plessmann U., Weber K. Neurofilament architecture combines structural principles of intermediate filaments with carboxy-terminal extensions increasing in size between triplet proteins. EMBO J. 1983;2(8):1295–1302. doi: 10.1002/j.1460-2075.1983.tb01584.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Geisler N., Kaufmann E., Weber K. Proteinchemical characterization of three structurally distinct domains along the protofilament unit of desmin 10 nm filaments. Cell. 1982 Aug;30(1):277–286. doi: 10.1016/0092-8674(82)90033-2. [DOI] [PubMed] [Google Scholar]
  15. Geisler N., Weber K. Comparison of the proteins of two immunologically distinct intermediate-sized filaments by amino acid sequence analysis: desmin and vimentin. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4120–4123. doi: 10.1073/pnas.78.7.4120. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Geisler N., Weber K. The amino acid sequence of chicken muscle desmin provides a common structural model for intermediate filament proteins. EMBO J. 1982;1(12):1649–1656. doi: 10.1002/j.1460-2075.1982.tb01368.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gigi O., Geiger B., Eshhar Z., Moll R., Schmid E., Winter S., Schiller D. L., Franke W. W. Detection of a cytokeratin determinant common to diverse epithelial cells by a broadly cross-reacting monoclonal antibody. EMBO J. 1982;1(11):1429–1437. doi: 10.1002/j.1460-2075.1982.tb01334.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Granger B. L., Lazarides E. Desmin and vimentin coexist at the periphery of the myofibril Z disc. Cell. 1979 Dec;18(4):1053–1063. doi: 10.1016/0092-8674(79)90218-6. [DOI] [PubMed] [Google Scholar]
  19. Hanukoglu I., Fuchs E. The cDNA sequence of a human epidermal keratin: divergence of sequence but conservation of structure among intermediate filament proteins. Cell. 1982 Nov;31(1):243–252. doi: 10.1016/0092-8674(82)90424-x. [DOI] [PubMed] [Google Scholar]
  20. Holtzer H., Bennett G. S., Tapscott S. J., Croop J. M., Toyama Y. Intermediate-size filaments: changes in synthesis and distribution in cells of the myogenic and neurogenic lineages. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 1):317–329. doi: 10.1101/sqb.1982.046.01.033. [DOI] [PubMed] [Google Scholar]
  21. Huiatt T. W., Robson R. M., Arakawa N., Stromer M. H. Desmin from avian smooth muscle. Purification and partial characterization. J Biol Chem. 1980 Jul 25;255(14):6981–6989. [PubMed] [Google Scholar]
  22. Ip W., Danto S. I., Fischman D. A. Detection of desmin-containing intermediate filaments in cultured muscle and nonmuscle cells by immunoelectron microscopy. J Cell Biol. 1983 Feb;96(2):401–408. doi: 10.1083/jcb.96.2.401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
  24. Lane E. B. Monoclonal antibodies provide specific intramolecular markers for the study of epithelial tonofilament organization. J Cell Biol. 1982 Mar;92(3):665–673. doi: 10.1083/jcb.92.3.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lazarides E., Hubbard B. D. Immunological characterization of the subunit of the 100 A filaments from muscle cells. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4344–4348. doi: 10.1073/pnas.73.12.4344. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lazarides E. Intermediate filaments--chemical heterogeneity in differentiation. Cell. 1981 Mar;23(3):649–650. doi: 10.1016/0092-8674(81)90427-x. [DOI] [PubMed] [Google Scholar]
  27. Lazarides E. Intermediate filaments: a chemically heterogeneous, developmentally regulated class of proteins. Annu Rev Biochem. 1982;51:219–250. doi: 10.1146/annurev.bi.51.070182.001251. [DOI] [PubMed] [Google Scholar]
  28. Miettinen M., Lehto V. P., Badley R. A., Virtanen I. Expression of intermediate filaments in soft-tissue sarcomas. Int J Cancer. 1982 Nov 15;30(5):541–546. doi: 10.1002/ijc.2910300502. [DOI] [PubMed] [Google Scholar]
  29. Moll R., Franke W. W., Schiller D. L., Geiger B., Krepler R. The catalog of human cytokeratins: patterns of expression in normal epithelia, tumors and cultured cells. Cell. 1982 Nov;31(1):11–24. doi: 10.1016/0092-8674(82)90400-7. [DOI] [PubMed] [Google Scholar]
  30. Osborn M., Caselitz J., Weber K. Heterogeneity of intermediate filament expression in vascular smooth muscle: a gradient in desmin positive cells from the rat aortic arch to the level of the arteria iliaca communis. Differentiation. 1981;20(3):196–202. doi: 10.1111/j.1432-0436.1981.tb01176.x. [DOI] [PubMed] [Google Scholar]
  31. Osborn M., Geisler N., Shaw G., Sharp G., Weber K. Intermediate filaments. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 1):413–429. doi: 10.1101/sqb.1982.046.01.040. [DOI] [PubMed] [Google Scholar]
  32. Osborn M., Geisler N., Shaw G., Sharp G., Weber K. Intermediate filaments. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 1):413–429. doi: 10.1101/sqb.1982.046.01.040. [DOI] [PubMed] [Google Scholar]
  33. Osborn M., Weber K. Tumor diagnosis by intermediate filament typing: a novel tool for surgical pathology. Lab Invest. 1983 Apr;48(4):372–394. [PubMed] [Google Scholar]
  34. Pruss R. M., Mirsky R., Raff M. C., Thorpe R., Dowding A. J., Anderton B. H. All classes of intermediate filaments share a common antigenic determinant defined by a monoclonal antibody. Cell. 1981 Dec;27(3 Pt 2):419–428. doi: 10.1016/0092-8674(81)90383-4. [DOI] [PubMed] [Google Scholar]
  35. Quax-Jeuken Y. E., Quax W. J., Bloemendal H. Primary and secondary structure of hamster vimentin predicted from the nucleotide sequence. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3548–3552. doi: 10.1073/pnas.80.12.3548. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Richardson F. L., Stromer M. H., Huiatt T. W., Robson R. M. Immunoelectron and immunofluorescence localization of desmin in mature avian muscles. Eur J Cell Biol. 1981 Dec;26(1):91–101. [PubMed] [Google Scholar]
  37. Small J. V., Sobieszek A. Studies on the function and composition of the 10-NM(100-A) filaments of vertebrate smooth muscle. J Cell Sci. 1977 Feb;23:243–268. doi: 10.1242/jcs.23.1.243. [DOI] [PubMed] [Google Scholar]
  38. Steinert P. M., Rice R. H., Roop D. R., Trus B. L., Steven A. C. Complete amino acid sequence of a mouse epidermal keratin subunit and implications for the structure of intermediate filaments. Nature. 1983 Apr 28;302(5911):794–800. doi: 10.1038/302794a0. [DOI] [PubMed] [Google Scholar]
  39. Stoeckel M. E., Osborn M., Porte A., Sacrez A., Batzenschlager A., Weber K. An unusual familial cardiomyopathy characterized by aberrant accumulations of desmin-type intermediate filaments. Virchows Arch A Pathol Anat Histol. 1981;393(1):53–60. doi: 10.1007/BF00430870. [DOI] [PubMed] [Google Scholar]
  40. Tokuyasu K. T. Visualization of longitudinally-oriented intermediate filaments in frozen sections of chicken cardiac muscle by a new staining method. J Cell Biol. 1983 Aug;97(2):562–565. doi: 10.1083/jcb.97.2.562. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. 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 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Travo P., Weber K., Osborn M. Co-existence of vimentin and desmin type intermediate filaments in a subpopulation of adult rat vascular smooth muscle cells growing in primary culture. Exp Cell Res. 1982 May;139(1):87–94. doi: 10.1016/0014-4827(82)90321-4. [DOI] [PubMed] [Google Scholar]
  43. Tseng S. C., Jarvinen M. J., Nelson W. G., Huang J. W., Woodcock-Mitchell J., Sun T. T. Correlation of specific keratins with different types of epithelial differentiation: monoclonal antibody studies. Cell. 1982 Sep;30(2):361–372. doi: 10.1016/0092-8674(82)90234-3. [DOI] [PubMed] [Google Scholar]
  44. Tuszynski G. P., Frank E. D., Damsky C. H., Buck C. A., Warren L. The detection of smooth muscle desmin-like protein in BHK21/C13 fibroblasts. J Biol Chem. 1979 Jul 10;254(13):6138–6143. [PubMed] [Google Scholar]

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