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. 1985 Apr;54(1):14–20. doi: 10.1128/jvi.54.1.14-20.1985

Neutralizing monoclonal antibodies to human rotavirus and indications of antigenic drift among strains from neonates.

B S Coulson, K J Fowler, R F Bishop, R G Cotton
PMCID: PMC254754  PMID: 2579249

Abstract

Cells producing neutralizing monoclonal antibodies to a serotype 3 human neonatal rotavirus strain RV-3 were derived by fusion of hyperimmunized mouse spleen cells with mouse myeloma cells. As ascites fluid, three rotavirus-neutralizing monoclonal antibodies were characterized by hemagglutination inhibition and reacted with 17 cultivable mammalian rotaviruses representing five virus serotypes, by fluorescent focus neutralization and enzyme immunoassay. Two antibodies, Mab RV-3:1 and Mab RV-3:2, reacted with the seven serotype 3 rotaviruses only. Mab RV-3:1 was shown to bind to the outer capsid glycoprotein gp34 of rotavirus when variants of SA 11 rotavirus were used, and it therefore appears to react with the major neutralization epitope of serotype 3 rotaviruses. The antibody Mab RV-3:3 was specific for an epitope of RV-3 rotavirus not present on any other rotavirus of any serotype tested, including another neonatal isolate of identical RNA electropherotype isolated from the same ward of the same hospital as RV-3 3 months earlier. These two viruses were also distinguishable by fluorescent focus neutralization, using antiserum to RV-3 virus. Western blot analysis showed binding of Mab RV-3:3 to the trypsin cleavage product of the outer capsid protein p86 of RV-3. This suggests that antigenic drift may have occurred among neonatal rotaviruses in Melbourne. These monoclonal antibodies will be useful in serotyping assays of rotaviruses directly in stool samples, and in further analysis of antigenic variation within the serotype.

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Selected References

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  1. Albert M. J., Bishop R. F. Cultivation of human rotaviruses in cell culture. J Med Virol. 1984;13(4):377–383. doi: 10.1002/jmv.1890130409. [DOI] [PubMed] [Google Scholar]
  2. Bastardo J. W., McKimm-Breschkin J. L., Sonza S., Mercer L. D., Holmes I. H. Preparation and characterization of antisera to electrophoretically purified SA11 virus polypeptides. Infect Immun. 1981 Dec;34(3):641–647. doi: 10.1128/iai.34.3.641-647.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beards G. M., Pilfold J. N., Thouless M. E., Flewett T. H. Rotavirus serotypes by serum neutralisation. J Med Virol. 1980;5(3):231–237. doi: 10.1002/jmv.1890050307. [DOI] [PubMed] [Google Scholar]
  4. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  5. Cohen J., Laporte J., Charpilienne A., Scherrer R. Activation of rotavirus RNA polymerase by calcium chelation. Arch Virol. 1979;60(3-4):177–186. doi: 10.1007/BF01317489. [DOI] [PubMed] [Google Scholar]
  6. Cotton R. G., Jennings I. G., Choo K. H., Fowler K. Isolation and characterization of a myeloma--spleen-cell hybrid producing antibody to phenylalanine hydroxylase. Biochem J. 1980 Dec 1;191(3):777–783. doi: 10.1042/bj1910777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Crainic R., Couillin P., Blondel B., Cabau N., Boué A., Horodniceanu F. Natural variation of poliovirus neutralization epitopes. Infect Immun. 1983 Sep;41(3):1217–1225. doi: 10.1128/iai.41.3.1217-1225.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dyall-Smith M. L., Holmes I. H. Sequence homology between human and animal rotavirus serotype-specific glycoproteins. Nucleic Acids Res. 1984 May 11;12(9):3973–3982. doi: 10.1093/nar/12.9.3973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Estes M. K., Graham D. Y., Mason B. B. Proteolytic enhancement of rotavirus infectivity: molecular mechanisms. J Virol. 1981 Sep;39(3):879–888. doi: 10.1128/jvi.39.3.879-888.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fulton R. W., Johnson C. A., Pearson N. J., Woode G. N. Isolation of a rotavirus from a newborn dog with diarrhea. Am J Vet Res. 1981 May;42(5):841–843. [PubMed] [Google Scholar]
  11. Gary G. W., Jr, Black D. R., Palmer E. Monoclonal IgG to the inner capsid of human rotavirus. Arch Virol. 1982;72(3):223–227. doi: 10.1007/BF01348968. [DOI] [PubMed] [Google Scholar]
  12. Gerna G., Passarani N., Battaglía M., Percivalle E. Rapid serotyping of human rotavirus strains by solid-phase immune electron microscopy. J Clin Microbiol. 1984 Feb;19(2):273–278. doi: 10.1128/jcm.19.2.273-278.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Greenberg H. B., Valdesuso J., van Wyke K., Midthun K., Walsh M., McAuliffe V., Wyatt R. G., Kalica A. R., Flores J., Hoshino Y. Production and preliminary characterization of monoclonal antibodies directed at two surface proteins of rhesus rotavirus. J Virol. 1983 Aug;47(2):267–275. doi: 10.1128/jvi.47.2.267-275.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Greenberg H., McAuliffe V., Valdesuso J., Wyatt R., Flores J., Kalica A., Hoshino Y., Singh N. Serological analysis of the subgroup protein of rotavirus, using monoclonal antibodies. Infect Immun. 1983 Jan;39(1):91–99. doi: 10.1128/iai.39.1.91-99.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Heinrich H. W. Untersuchungen zum Nachweis einer experimentell provozierten Antigendrift bei Rotaviren. Arch Exp Veterinarmed. 1983 Sep;37(5):667–670. [PubMed] [Google Scholar]
  16. Kalica A. R., Flores J., Greenberg H. B. Identification of the rotaviral gene that codes for hemagglutination and protease-enhanced plaque formation. Virology. 1983 Feb;125(1):194–205. doi: 10.1016/0042-6822(83)90073-9. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Matsuno S., Inouye S., Kono R. Plaque assay of neonatal calf diarrhea virus and the neutralizing antibody in human sera. J Clin Microbiol. 1977 Jan;5(1):1–4. doi: 10.1128/jcm.5.1.1-4.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. McLean B., Holmes I. H. Transfer of antirotaviral antibodies from mothers to their infants. J Clin Microbiol. 1980 Sep;12(3):320–325. doi: 10.1128/jcm.12.3.320-325.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. McLean B., Sonza S., Holmes I. H. Measurement of immunoglobulin A, G, and M class rotavirus antibodies in serum and mucosal secretions. J Clin Microbiol. 1980 Sep;12(3):314–319. doi: 10.1128/jcm.12.3.314-319.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rodger S. M., Bishop R. F., Birch C., McLean B., Holmes I. H. Molecular epidemiology of human rotaviruses in Melbourne, Australia, from 1973 to 1979, as determined by electrophoresis of genome ribonucleic acid. J Clin Microbiol. 1981 Feb;13(2):272–278. doi: 10.1128/jcm.13.2.272-278.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rodger S. M., Schnagl R. D., Holmes I. H. Biochemical and biophysical characteristics of diarrhea viruses of human and calf origin. J Virol. 1975 Nov;16(5):1229–1235. doi: 10.1128/jvi.16.5.1229-1235.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Roseto A., Scherrer R., Cohen J., Guillemin M. C., Charpilienne A., Feynerol C., Peries J. Isolation and characterization of anti-rotavirus immunoglobulins secreted by cloned hybridoma cell lines. J Gen Virol. 1983 Jan;64(Pt 1):237–240. doi: 10.1099/0022-1317-64-1-237. [DOI] [PubMed] [Google Scholar]
  24. Sato K., Inaba Y., Shinozaki T., Fujii R., Matumoto M. Isolation of human rotavirus in cell cultures: brief report. Arch Virol. 1981;69(2):155–160. doi: 10.1007/BF01315159. [DOI] [PubMed] [Google Scholar]
  25. Smith M. L., Lazdins I., Holmes I. H. Coding assignments of double-stranded RNA segments of SA 11 rotavirus established by in vitro translation. J Virol. 1980 Mar;33(3):976–982. doi: 10.1128/jvi.33.3.976-982.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sonza S., Breschkin A. M., Holmes I. H. Derivation of neutralizing monoclonal antibodies against rotavirus. J Virol. 1983 Mar;45(3):1143–1146. doi: 10.1128/jvi.45.3.1143-1146.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sonza S., Breschkin A. M., Holmes I. H. The major surface glycoprotein of simian rotavirus (SA11) contains distinct epitopes. Virology. 1984 Apr 30;134(2):318–327. doi: 10.1016/0042-6822(84)90300-3. [DOI] [PubMed] [Google Scholar]
  28. Thouless M. E., Beards G. M., Flewett T. H. Serotyping and subgrouping of rotavirus strains by the ELISA test. Arch Virol. 1982;73(3-4):219–230. doi: 10.1007/BF01318076. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Urasawa S., Urasawa T., Taniguchi K. Three human rotavirus serotypes demonstrated by plaque neutralization of isolated strains. Infect Immun. 1982 Nov;38(2):781–784. doi: 10.1128/iai.38.2.781-784.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Woode G. N., Bridger J. C., Hall G., Dennis M. J. The isolation of a reovirus-like agent associated with diarrhoea in colostrum-deprived calves in Great Britain. Res Vet Sci. 1974 Jan;16(1):102–105. [PubMed] [Google Scholar]
  32. Wyatt R. G., James H. D., Jr, Pittman A. L., Hoshino Y., Greenberg H. B., Kalica A. R., Flores J., Kapikian A. Z. Direct isolation in cell culture of human rotaviruses and their characterization into four serotypes. J Clin Microbiol. 1983 Aug;18(2):310–317. doi: 10.1128/jcm.18.2.310-317.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Yewdell J. W., Webster R. G., Gerhard W. U. Antigenic variation in three distinct determinants of an influenza type A haemagglutinin molecule. Nature. 1979 May 17;279(5710):246–248. doi: 10.1038/279246a0. [DOI] [PubMed] [Google Scholar]

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