Abstract
Astroviruses are important agents of pediatric gastroenteritis. To better understand astrovirus antigenic structure and the basis of protective immunity, monoclonal antibodies (MAbs) were produced against serotype 1 human astrovirus. Four MAbs were generated. One MAb (8G4) was nonneutralizing but reacted to all seven serotypes of astrovirus by enzyme-linked immunosorbentassay (ELISA) and immunoperoxidase staining of infected cells. Three MAbs were found to have potent neutralizing activity against astrovirus. The first (5B7) was serotype 1 specific, another (7C2) neutralized all seven human astrovirus serotypes, while the third (3B2) neutralized serotypes 1 and 7. Immunoprecipitation of radiolabeled astrovirus proteins from supernatants of astrovirus-infected cells showed that all three neutralizing antibodies reacted with VP29. MAb 5B7 also reacted strongly with VP26. A competition ELISA showed that all three neutralizing antibodies competed with each other for binding to purified astrovirus virions, suggesting that their epitopes were topographically in close proximity. None of the neutralizing MAbs competed with nonneutralizing MAb 8G4. The neutralizing MAbs were used to select antigenic variant astroviruses, which were then studied in neutralization assays. These assays also suggested a close relationship between the respective epitopes. All three neutralizing MAbs were able to prevent attachment of radiolabeled astrovirus particles to human Caco 2 intestinal cell monolayers. Taken together, these data suggest that the astrovirus capsid protein VP29 may be important in viral neutralization, heterotypic immunity, and virus attachment to target cells.
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Selected References
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- Carter M. J., Willcocks M. M. The molecular biology of astroviruses. Arch Virol Suppl. 1996;12:277–285. doi: 10.1007/978-3-7091-6553-9_30. [DOI] [PubMed] [Google Scholar]
- Glass R. I., Noel J., Mitchell D., Herrmann J. E., Blacklow N. R., Pickering L. K., Dennehy P., Ruiz-Palacios G., de Guerrero M. L., Monroe S. S. The changing epidemiology of astrovirus-associated gastroenteritis: a review. Arch Virol Suppl. 1996;12:287–300. doi: 10.1007/978-3-7091-6553-9_31. [DOI] [PubMed] [Google Scholar]
- Greenberg H. B., Matsui S. M. Astroviruses and caliciviruses: emerging enteric pathogens. Infect Agents Dis. 1992 Apr;1(2):71–91. [PubMed] [Google Scholar]
- 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]
- Herrmann J. E., Hudson R. W., Perron-Henry D. M., Kurtz J. B., Blacklow N. R. Antigenic characterization of cell-cultivated astrovirus serotypes and development of astrovirus-specific monoclonal antibodies. J Infect Dis. 1988 Jul;158(1):182–185. doi: 10.1093/infdis/158.1.182. [DOI] [PubMed] [Google Scholar]
- Kriston S., Willcocks M. M., Carter M. J., Cubitt W. D. Seroprevalence of astrovirus types 1 and 6 in London, determined using recombinant virus antigen. Epidemiol Infect. 1996 Aug;117(1):159–164. doi: 10.1017/s0950268800001266. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kurtz J. B., Lee T. W. Human astrovirus serotypes. Lancet. 1984 Dec 15;2(8416):1405–1405. doi: 10.1016/s0140-6736(84)92101-9. [DOI] [PubMed] [Google Scholar]
- Lee T. W., Kurtz J. B. Prevalence of human astrovirus serotypes in the Oxford region 1976-92, with evidence for two new serotypes. Epidemiol Infect. 1994 Feb;112(1):187–193. doi: 10.1017/s0950268800057551. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Midthun K., Greenberg H. B., Kurtz J. B., Gary G. W., Lin F. Y., Kapikian A. Z. Characterization and seroepidemiology of a type 5 astrovirus associated with an outbreak of gastroenteritis in Marin County, California. J Clin Microbiol. 1993 Apr;31(4):955–962. doi: 10.1128/jcm.31.4.955-962.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Monroe S. S., Jiang B., Stine S. E., Koopmans M., Glass R. I. Subgenomic RNA sequence of human astrovirus supports classification of Astroviridae as a new family of RNA viruses. J Virol. 1993 Jun;67(6):3611–3614. doi: 10.1128/jvi.67.6.3611-3614.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Monroe S. S., Stine S. E., Gorelkin L., Herrmann J. E., Blacklow N. R., Glass R. I. Temporal synthesis of proteins and RNAs during human astrovirus infection of cultured cells. J Virol. 1991 Feb;65(2):641–648. doi: 10.1128/jvi.65.2.641-648.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Noel J. S., Lee T. W., Kurtz J. B., Glass R. I., Monroe S. S. Typing of human astroviruses from clinical isolates by enzyme immunoassay and nucleotide sequencing. J Clin Microbiol. 1995 Apr;33(4):797–801. doi: 10.1128/jcm.33.4.797-801.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Noel J., Cubitt D. Identification of astrovirus serotypes from children treated at the Hospitals for Sick Children, London 1981-93. Epidemiol Infect. 1994 Aug;113(1):153–159. doi: 10.1017/s0950268800051578. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ruggeri F. M., Greenberg H. B. Antibodies to the trypsin cleavage peptide VP8 neutralize rotavirus by inhibiting binding of virions to target cells in culture. J Virol. 1991 May;65(5):2211–2219. doi: 10.1128/jvi.65.5.2211-2219.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sanchez-Fauquier A., Carrascosa A. L., Carrascosa J. L., Otero A., Glass R. I., Lopez J. A., San Martin C., Melero J. A. Characterization of a human astrovirus serotype 2 structural protein (VP26) that contains an epitope involved in virus neutralization. Virology. 1994 Jun;201(2):312–320. doi: 10.1006/viro.1994.1296. [DOI] [PubMed] [Google Scholar]
- Shaw R. D., Vo P. T., Offit P. A., Coulson B. S., Greenberg H. B. Antigenic mapping of the surface proteins of rhesus rotavirus. Virology. 1986 Dec;155(2):434–451. doi: 10.1016/0042-6822(86)90205-9. [DOI] [PubMed] [Google Scholar]
- Willcocks M. M., Carter M. J., Laidler F. R., Madeley C. R. Growth and characterisation of human faecal astrovirus in a continuous cell line. Arch Virol. 1990;113(1-2):73–81. doi: 10.1007/BF01318354. [DOI] [PubMed] [Google Scholar]