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. 1982 Jan;35(1):179–186. doi: 10.1128/iai.35.1.179-186.1982

Differentiation of mumps virus strains with monoclonal antibody to the HN glycoprotein.

A C Server, D C Merz, M N Waxham, J S Wolinsky
PMCID: PMC351013  PMID: 6172379

Abstract

A hybridoma cell line secreting antibody of the immunoglobulin G3 isotype with kappa light chains and with activity against the HN glycoprotein of the Kilham strain of mumps virus was established. The antibody exhibited structural homogeneity in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and had a microheterogeneous isoelectric spectrum characteristic of an antibody of monoclonal origin. The specificity of the monoclonal antibody, shown by immunoprecipitation performed with radiolabeled virus and infected cell lysates, was for the larger mumps virus glycoprotein. In functional assays the antibody inhibited the hemagglutinating and neuraminidase activities and neutralized the infectivity of the homologous Kilham strain of virus and clearly differentiated this strain from two heterologous strains, Enders and O'Take. The antibody was markedly less effective with the O'Take strain than with either the Kilham or Enders strain in inhibiting both hemagglutination and neuraminidase activity against the macromolecular substrate fetuin. The inhibition of the neuraminidase activity of the Kilham strain was independent of substrate size, the antibody inhibiting the hydrolysis of both fetuin and the trisaccharide neuraminlactose. By contrast, the antibody did not inhibit the hydrolysis of neuraminlactose by the two heterologous mumps strains. These results provide the first demonstration of antigenic differences between mumps virus strains and highlight the utility of monoclonal antibody in analyzing the structural basis underlying functional activities of the HN glycoproteins.

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Selected References

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  1. ADA G. L., LIND P. E. AN IMMUNOLOGICAL STUDY OF AVIAN, VIRAL AND BACTERIAL NEURAMINIDASE BASED ON SPECIFIC INHIBITION OF ENZYME BY ANTIBODY. J Gen Microbiol. 1963 Aug;32:225–233. doi: 10.1099/00221287-32-2-225. [DOI] [PubMed] [Google Scholar]
  2. AMINOFF D. Methods for the quantitative estimation of N-acetylneuraminic acid and their application to hydrolysates of sialomucoids. Biochem J. 1961 Nov;81:384–392. doi: 10.1042/bj0810384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  4. CABASSO V. J., COX H. R. Hemagglutination behavior of mumps virus strains of different origin. Proc Soc Exp Biol Med. 1955 Mar;88(3):370–372. doi: 10.3181/00379727-88-21591. [DOI] [PubMed] [Google Scholar]
  5. CHALLUT F., CHARDONNET Y., FAVIER G., SOHIER R. Etude du pouvoir hémagglutinant de neuf souches de virus des oreillons pour des hématies de différentes espéces. Ann Inst Pasteur (Paris) 1956 Aug;91(2):137–146. [PubMed] [Google Scholar]
  6. Coyle P. K., Wolinsky J. S. Characterization of immune complexes in progressive rubella panencephalitis. Ann Neurol. 1981 Jun;9(6):557–562. doi: 10.1002/ana.410090608. [DOI] [PubMed] [Google Scholar]
  7. Garten W., Berk W., Nagai Y., Rott R., Klenk H. D. Mutational changes of the protease susceptibility of glycoprotein F of Newcastle disease virus: effects on pathogenicity. J Gen Virol. 1980 Sep;50(1):135–147. doi: 10.1099/0022-1317-50-1-135. [DOI] [PubMed] [Google Scholar]
  8. Glathe H., Nöbel B. Biologische Eigenschaften des Mumpsvirus. Das Verhalte im T50-Marker. J Hyg Epidemiol Microbiol Immunol. 1977;21(2):162–168. [PubMed] [Google Scholar]
  9. Hardwick J. M., Bussell R. H. Disulfide bonds in Sendai virus glycoproteins. Intervirology. 1979;11(5):300–306. doi: 10.1159/000149049. [DOI] [PubMed] [Google Scholar]
  10. Hewitt J. A., Allen G. Controlled proteolytic digestion of the M-protein of Sendai virus: the isolation of a fragment of 30000 molecular weight. J Gen Virol. 1978 May;39(2):311–319. doi: 10.1099/0022-1317-39-2-311. [DOI] [PubMed] [Google Scholar]
  11. Jensik S. C., Silver S. Polypeptides of mumps virus. J Virol. 1976 Feb;17(2):363–373. doi: 10.1128/jvi.17.2.363-373.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kearney J. F., Radbruch A., Liesegang B., Rajewsky K. A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. J Immunol. 1979 Oct;123(4):1548–1550. [PubMed] [Google Scholar]
  13. Klamm H. Some properties of mumps virus neuraminidase. Acta Virol. 1980 Mar;24(2):127–131. [PubMed] [Google Scholar]
  14. Lamb R. A., Choppin P. W. The synthesis of Sendai virus polypeptides in infected cells. III. Phosphorylation of polypeptides. Virology. 1977 Sep;81(2):382–397. doi: 10.1016/0042-6822(77)90154-4. [DOI] [PubMed] [Google Scholar]
  15. Lamb R. A., Mahy B. W., Choppin P. W. The synthesis of sendai virus polypeptides in infected cells. Virology. 1976 Jan;69(1):116–131. doi: 10.1016/0042-6822(76)90199-9. [DOI] [PubMed] [Google Scholar]
  16. Leprat R., Aymard M. Selective inactivation of hemagglutinin and neuraminidase on mumps virus. Arch Virol. 1979;61(4):273–281. doi: 10.1007/BF01315013. [DOI] [PubMed] [Google Scholar]
  17. Markwell M. A., Haas S. M., Bieber L. L., Tolbert N. E. A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem. 1978 Jun 15;87(1):206–210. doi: 10.1016/0003-2697(78)90586-9. [DOI] [PubMed] [Google Scholar]
  18. McCarthy M., Johnson R. T. A comparison of the structural polypeptides of five strains of mumps virus. J Gen Virol. 1980 Jan;46(1):15–27. doi: 10.1099/0022-1317-46-1-15. [DOI] [PubMed] [Google Scholar]
  19. Merz D. C., Prehm P., Scheid A., Choppin P. W. Inhibition of the neuraminidase of paramyxoviruses by halide ions: a possible means of modulating the two activities of the HN protein. Virology. 1981 Jul 15;112(1):296–305. doi: 10.1016/0042-6822(81)90635-8. [DOI] [PubMed] [Google Scholar]
  20. Merz D. C., Scheid A., Choppin P. W. Immunological studies of the functions of paramyxovirus glycoproteins. Virology. 1981 Feb;109(1):94–105. doi: 10.1016/0042-6822(81)90474-8. [DOI] [PubMed] [Google Scholar]
  21. Merz D. C., Wolinsky J. S. Biochemical features of mumps virus neuraminidases and their relationship with pathogenicity. Virology. 1981 Oct 15;114(1):218–227. doi: 10.1016/0042-6822(81)90267-1. [DOI] [PubMed] [Google Scholar]
  22. Orvell C. Immunological properties of purified mumps virus glycoproteins. J Gen Virol. 1978 Dec;41(3):517–526. doi: 10.1099/0022-1317-41-3-517. [DOI] [PubMed] [Google Scholar]
  23. Rima B. K., Roberts M. W., McAdam W. D., Martin S. J. Polypeptide synthesis i mumps virus-infected cells. J Gen Virol. 1980 Feb;46(2):501–505. doi: 10.1099/0022-1317-46-2-501. [DOI] [PubMed] [Google Scholar]
  24. Roehrig J. T., Corser J. A., Schlesinger M. J. Isolation and characterization of hybrid cell lines producing monoclonal antibodies directed against the structural proteins of Sindbis virus. Virology. 1980 Feb;101(1):41–49. doi: 10.1016/0042-6822(80)90481-x. [DOI] [PubMed] [Google Scholar]
  25. Scheid A., Choppin P. W. Identification of biological activities of paramyxovirus glycoproteins. Activation of cell fusion, hemolysis, and infectivity of proteolytic cleavage of an inactive precursor protein of Sendai virus. Virology. 1974 Feb;57(2):475–490. doi: 10.1016/0042-6822(74)90187-1. [DOI] [PubMed] [Google Scholar]
  26. Scheid A., Choppin P. W. The hemagglutinating and neuraminidase protein of a paramyxovirus: interaction with neuraminic acid in affinity chromatography. Virology. 1974 Nov;62(1):125–133. doi: 10.1016/0042-6822(74)90308-0. [DOI] [PubMed] [Google Scholar]
  27. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  28. Williamson A. R., Salaman M. R., Kreth H. W. Microheterogeneity and allomorphism of proteins. Ann N Y Acad Sci. 1973 Jun 15;209:210–224. doi: 10.1111/j.1749-6632.1973.tb47530.x. [DOI] [PubMed] [Google Scholar]
  29. Wolinsky J. S., Baringer J. R., Margolis G., Kilham L. Ultrastructure of mumps virus replication in newborn hamster central nervous system. Lab Invest. 1974 Oct;31(4):403–412. [PubMed] [Google Scholar]

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