Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1991 Dec;65(12):7037–7040. doi: 10.1128/jvi.65.12.7037-7040.1991

Use of a lambda gt11 expression library to localize a neutralizing antibody-binding site in glycoprotein E2 of Sindbis virus.

K S Wang 1, J H Strauss 1
PMCID: PMC250823  PMID: 1719239

Abstract

The Sindbis virus envelope contains two species of integral membrane glycoproteins, E1 and E2. These proteins form heterodimers, and three dimeric units assemble to form spikes incorporated into the viral surface which play an important role in the specific attachment of Sindbis virus to host cells. To map the neutralization epitopes on the surface of the virus, we constructed a lambda gt11 expression library with cDNA inserts 100 to 300 nucleotides long obtained from randomly primed synthesis on Sindbis virus genomic RNA. This library was screened with five different neutralizing monoclonal antibodies (MAbs) specific for E2 (MAbs 50, 51, 49, 18, and 23) and with one neutralizing MAb specific for E1 (MAb 33). When 10(6) lambda gt11 plaques were screened with each antibody, four positive clones that reacted with E2-specific MAb 23 were found. These four clones contained overlapping inserts from glycoprotein E2; the domain from residues 173 to 220 of glycoprotein E2 was present in all inserts, and we concluded that this region contains the neutralization epitope recognized by the antibody. No clones that reacted with the other antibodies examined were found, and we concluded that these antibodies probably recognize conformational epitopes not present in the lambda gt11 library. We suggest that the E2 domain from residues 173 to 220 is a major antigenic determinant of Sindbis virus and that this domain is important for virus attachment to cells.

Full text

PDF
7037

Images in this article

7038Image
on p.7038

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bell J. R., Strauss E. G., Strauss J. H. Purification and amino acid compositions of the structural proteins of sindbis virus. Virology. 1979 Sep;97(2):287–294. doi: 10.1016/0042-6822(79)90340-4. [DOI] [PubMed] [Google Scholar]
  2. Chanas A. C., Johnson B. K., Simpson D. I. Antigenic relationships of alphaviruses by a simple micro-culture cross-neutralization method. J Gen Virol. 1976 Aug;32(2):295–300. doi: 10.1099/0022-1317-32-2-295. [DOI] [PubMed] [Google Scholar]
  3. Co M. S., Gaulton G. N., Fields B. N., Greene M. I. Isolation and biochemical characterization of the mammalian reovirus type 3 cell-surface receptor. Proc Natl Acad Sci U S A. 1985 Mar;82(5):1494–1498. doi: 10.1073/pnas.82.5.1494. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Davis N. L., Pence D. F., Meyer W. J., Schmaljohn A. L., Johnston R. E. Alternative forms of a strain-specific neutralizing antigenic site on the Sindbis virus E2 glycoprotein. Virology. 1987 Nov;161(1):101–108. doi: 10.1016/0042-6822(87)90175-9. [DOI] [PubMed] [Google Scholar]
  5. Diamond D. C., Jameson B. A., Bonin J., Kohara M., Abe S., Itoh H., Komatsu T., Arita M., Kuge S., Nomoto A. Antigenic variation and resistance to neutralization in poliovirus type 1. Science. 1985 Sep 13;229(4718):1090–1093. doi: 10.1126/science.2412292. [DOI] [PubMed] [Google Scholar]
  6. Gaulton G. N., Greene M. I. Idiotypic mimicry of biological receptors. Annu Rev Immunol. 1986;4:253–280. doi: 10.1146/annurev.iy.04.040186.001345. [DOI] [PubMed] [Google Scholar]
  7. Hsu M. T., Kung H. J., Davidson N. An electron microscope study of Sindbis virus RNA. Cold Spring Harb Symp Quant Biol. 1974;38:943–950. doi: 10.1101/sqb.1974.038.01.096. [DOI] [PubMed] [Google Scholar]
  8. Johnson B. J., Brubaker J. R., Roehrig J. T., Trent D. W. Variants of Venezuelan equine encephalitis virus that resist neutralization define a domain of the E2 glycoprotein. Virology. 1990 Aug;177(2):676–683. doi: 10.1016/0042-6822(90)90533-w. [DOI] [PubMed] [Google Scholar]
  9. Laver W. G., Air G. M., Webster R. G., Smith-Gill S. J. Epitopes on protein antigens: misconceptions and realities. Cell. 1990 May 18;61(4):553–556. doi: 10.1016/0092-8674(90)90464-p. [DOI] [PubMed] [Google Scholar]
  10. Lerner R. A. Tapping the immunological repertoire to produce antibodies of predetermined specificity. Nature. 1982 Oct 14;299(5884):593–596. doi: 10.1038/299592a0. [DOI] [PubMed] [Google Scholar]
  11. Lobigs M., Dalgarno L., Schlesinger J. J., Weir R. C. Location of a neutralization determinant in the E protein of yellow fever virus (17D vaccine strain). Virology. 1987 Dec;161(2):474–478. doi: 10.1016/0042-6822(87)90141-3. [DOI] [PubMed] [Google Scholar]
  12. Lustig S., Jackson A. C., Hahn C. S., Griffin D. E., Strauss E. G., Strauss J. H. Molecular basis of Sindbis virus neurovirulence in mice. J Virol. 1988 Jul;62(7):2329–2336. doi: 10.1128/jvi.62.7.2329-2336.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Parry N., Fox G., Rowlands D., Brown F., Fry E., Acharya R., Logan D., Stuart D. Structural and serological evidence for a novel mechanism of antigenic variation in foot-and-mouth disease virus. Nature. 1990 Oct 11;347(6293):569–572. doi: 10.1038/347569a0. [DOI] [PubMed] [Google Scholar]
  14. Pence D. F., Davis N. L., Johnston R. E. Antigenic and genetic characterization of Sindbis virus monoclonal antibody escape mutants which define a pathogenesis domain on glycoprotein E2. Virology. 1990 Mar;175(1):41–49. doi: 10.1016/0042-6822(90)90184-s. [DOI] [PubMed] [Google Scholar]
  15. Pierce J. S., Strauss E. G., Strauss J. H. Effect of ionic strength on the binding of Sindbis virus to chick cells. J Virol. 1974 May;13(5):1030–1036. doi: 10.1128/jvi.13.5.1030-1036.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Roehrig J. T., Gorski D., Schlesinger M. J. Properties of monoclonal antibodies directed against the glycoproteins of Sindbis virus. J Gen Virol. 1982 Apr;59(Pt 2):421–425. doi: 10.1099/0022-1317-59-2-421. [DOI] [PubMed] [Google Scholar]
  17. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Schmaljohn A. L., Johnson E. D., Dalrymple J. M., Cole G. A. Non-neutralizing monoclonal antibodies can prevent lethal alphavirus encephalitis. Nature. 1982 May 6;297(5861):70–72. doi: 10.1038/297070a0. [DOI] [PubMed] [Google Scholar]
  19. Schmaljohn A. L., Kokubun K. M., Cole G. A. Protective monoclonal antibodies define maturational and pH-dependent antigenic changes in Sindbis virus E1 glycoprotein. Virology. 1983 Oct 15;130(1):144–154. doi: 10.1016/0042-6822(83)90124-1. [DOI] [PubMed] [Google Scholar]
  20. Sherry B., Mosser A. G., Colonno R. J., Rueckert R. R. Use of monoclonal antibodies to identify four neutralization immunogens on a common cold picornavirus, human rhinovirus 14. J Virol. 1986 Jan;57(1):246–257. doi: 10.1128/jvi.57.1.246-257.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Stanley J., Cooper S. J., Griffin D. E. Alphavirus neurovirulence: monoclonal antibodies discriminating wild-type from neuroadapted Sindbis virus. J Virol. 1985 Oct;56(1):110–119. doi: 10.1128/jvi.56.1.110-119.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Stec D. S., Waddell A., Schmaljohn C. S., Cole G. A., Schmaljohn A. L. Antibody-selected variation and reversion in Sindbis virus neutralization epitopes. J Virol. 1986 Mar;57(3):715–720. doi: 10.1128/jvi.57.3.715-720.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Strauss E. G., Stec D. S., Schmaljohn A. L., Strauss J. H. Identification of antigenically important domains in the glycoproteins of Sindbis virus by analysis of antibody escape variants. J Virol. 1991 Sep;65(9):4654–4664. doi: 10.1128/jvi.65.9.4654-4664.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tucker P. C., Griffin D. E. Mechanism of altered Sindbis virus neurovirulence associated with a single-amino-acid change in the E2 Glycoprotein. J Virol. 1991 Mar;65(3):1551–1557. doi: 10.1128/jvi.65.3.1551-1557.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Vrati S., Fernon C. A., Dalgarno L., Weir R. C. Location of a major antigenic site involved in Ross River virus neutralization. Virology. 1988 Feb;162(2):346–353. doi: 10.1016/0042-6822(88)90474-6. [DOI] [PubMed] [Google Scholar]
  26. Wang K. S., Schmaljohn A. L., Kuhn R. J., Strauss J. H. Antiidiotypic antibodies as probes for the Sindbis virus receptor. Virology. 1991 Apr;181(2):694–702. doi: 10.1016/0042-6822(91)90903-o. [DOI] [PubMed] [Google Scholar]
  27. Wiley D. C., Skehel J. J. The structure and function of the hemagglutinin membrane glycoprotein of influenza virus. Annu Rev Biochem. 1987;56:365–394. doi: 10.1146/annurev.bi.56.070187.002053. [DOI] [PubMed] [Google Scholar]
  28. Young R. A., Davis R. W. Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1194–1198. doi: 10.1073/pnas.80.5.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES