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. 1992 Sep;66(9):5290–5297. doi: 10.1128/jvi.66.9.5290-5297.1992

A continuous sequence of more than 70 amino acids is essential for antibody binding to the dominant antigenic site of glycoprotein gp58 of human cytomegalovirus.

B Wagner 1, B Kropff 1, H Kalbacher 1, W Britt 1, V A Sundqvist 1, L Ostberg 1, M Mach 1
PMCID: PMC289083  PMID: 1323695

Abstract

Antigenic domain 1 (AD-1) on glycoprotein gp58 of human cytomegalovirus was characterized in detail, using mouse and human monoclonal antibodies as well as human convalescent sera. Series of procaryotically expressed fusion proteins and synthetic peptides of various lengths were used as sources of antigen. Binding of antibodies was found to depend on a continuous sequence of more than 70 amino acids between residues 552 and 635 of gp58. The fine specificities for sequences involved in antibody binding were (i) amino acids 557 to 635 for neutralizing as well as nonneutralizing mouse monoclonal antibodies, (ii) amino acids 552 to 630 for a neutralizing human monoclonal antibody, and (iii) amino acids 557 to 630 for antibodies present in human sera. Experiments involving fragments of AD-1, presented either as procaryotically expressed fusion protein or as synthetic peptides, indicated that the intact structure was required for recognition of AD-1 by antibodies.

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Selected References

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  1. Alford C. A., Hayes K., Britt W. Primary cytomegalovirus infection in pregnancy: comparison of antibody responses to virus-encoded proteins between women with and without intrauterine infection. J Infect Dis. 1988 Nov;158(5):917–924. doi: 10.1093/infdis/158.5.917. [DOI] [PubMed] [Google Scholar]
  2. Andreoni M., Faircloth M., Vugler L., Britt W. J. A rapid microneutralization assay for the measurement of neutralizing antibody reactive with human cytomegalovirus. J Virol Methods. 1989 Feb;23(2):157–167. doi: 10.1016/0166-0934(89)90129-8. [DOI] [PubMed] [Google Scholar]
  3. Britt W. J., Auger D. Synthesis and processing of the envelope gp55-116 complex of human cytomegalovirus. J Virol. 1986 Apr;58(1):185–191. doi: 10.1128/jvi.58.1.185-191.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Britt W. J. Neutralizing antibodies detect a disulfide-linked glycoprotein complex within the envelope of human cytomegalovirus. Virology. 1984 Jun;135(2):369–378. doi: 10.1016/0042-6822(84)90193-4. [DOI] [PubMed] [Google Scholar]
  5. Britt W. J., Vugler L. G. Processing of the gp55-116 envelope glycoprotein complex (gB) of human cytomegalovirus. J Virol. 1989 Jan;63(1):403–410. doi: 10.1128/jvi.63.1.403-410.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Britt W. J., Vugler L., Butfiloski E. J., Stephens E. B. Cell surface expression of human cytomegalovirus (HCMV) gp55-116 (gB): use of HCMV-recombinant vaccinia virus-infected cells in analysis of the human neutralizing antibody response. J Virol. 1990 Mar;64(3):1079–1085. doi: 10.1128/jvi.64.3.1079-1085.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brown L. E., Murray J. M., Anders E. M., Tang X. L., White D. O., Tregear G. W., Jackson D. C. Genetic control and fine specificity of the immune response to a synthetic peptide of influenza virus hemagglutinin. J Virol. 1988 May;62(5):1746–1752. doi: 10.1128/jvi.62.5.1746-1752.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chee M. S., Bankier A. T., Beck S., Bohni R., Brown C. M., Cerny R., Horsnell T., Hutchison C. A., 3rd, Kouzarides T., Martignetti J. A. Analysis of the protein-coding content of the sequence of human cytomegalovirus strain AD169. Curr Top Microbiol Immunol. 1990;154:125–169. doi: 10.1007/978-3-642-74980-3_6. [DOI] [PubMed] [Google Scholar]
  9. Chou S. W., Dennison K. M. Analysis of interstrain variation in cytomegalovirus glycoprotein B sequences encoding neutralization-related epitopes. J Infect Dis. 1991 Jun;163(6):1229–1234. doi: 10.1093/infdis/163.6.1229. [DOI] [PubMed] [Google Scholar]
  10. Cranage M. P., Kouzarides T., Bankier A. T., Satchwell S., Weston K., Tomlinson P., Barrell B., Hart H., Bell S. E., Minson A. C. Identification of the human cytomegalovirus glycoprotein B gene and induction of neutralizing antibodies via its expression in recombinant vaccinia virus. EMBO J. 1986 Nov;5(11):3057–3063. doi: 10.1002/j.1460-2075.1986.tb04606.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Darlington J., Super M., Patel K., Grundy J. E., Griffiths P. D., Emery V. C. Use of the polymerase chain reaction to analyse sequence variation within a major neutralizing epitope of glycoprotein B (gp58) in clinical isolates of human cytomegalovirus. J Gen Virol. 1991 Aug;72(Pt 8):1985–1989. doi: 10.1099/0022-1317-72-8-1985. [DOI] [PubMed] [Google Scholar]
  12. Ehrlich P. H., Harfeldt K. E., Justice J. C., Moustafa Z. A., Ostberg L. Rhesus monkey responses to multiple injections of human monoclonal antibodies. Hybridoma. 1987 Apr;6(2):151–160. doi: 10.1089/hyb.1987.6.151. [DOI] [PubMed] [Google Scholar]
  13. Ellinger S., Mach M., Korn K., Jahn G. Cleavage and purification of prokaryotically expressed HIV gag and env fusion proteins for detection of HIV antibodies in the ELISA. Virology. 1991 Feb;180(2):811–813. doi: 10.1016/0042-6822(91)90097-u. [DOI] [PubMed] [Google Scholar]
  14. Farrar G. H., Greenaway P. J. Characterization of glycoprotein complexes present in human cytomegalovirus envelopes. J Gen Virol. 1986 Jul;67(Pt 7):1469–1473. doi: 10.1099/0022-1317-67-7-1469. [DOI] [PubMed] [Google Scholar]
  15. Gretch D. R., Gehrz R. C., Stinski M. F. Characterization of a human cytomegalovirus glycoprotein complex (gcI). J Gen Virol. 1988 Jun;69(Pt 6):1205–1215. doi: 10.1099/0022-1317-69-6-1205. [DOI] [PubMed] [Google Scholar]
  16. Gretch D. R., Kari B., Rasmussen L., Gehrz R. C., Stinski M. F. Identification and characterization of three distinct families of glycoprotein complexes in the envelopes of human cytomegalovirus. J Virol. 1988 Mar;62(3):875–881. doi: 10.1128/jvi.62.3.875-881.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gönczöl E., Plotkin S. Progress in vaccine development for prevention of human cytomegalovirus infection. Curr Top Microbiol Immunol. 1990;154:255–274. doi: 10.1007/978-3-642-74980-3_10. [DOI] [PubMed] [Google Scholar]
  18. Houghten R. A. General method for the rapid solid-phase synthesis of large numbers of peptides: specificity of antigen-antibody interaction at the level of individual amino acids. Proc Natl Acad Sci U S A. 1985 Aug;82(15):5131–5135. doi: 10.1073/pnas.82.15.5131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kari B., Gehrz R. Biochemical and immunological analysis of discontinuous epitopes in the family of human cytomegalovirus glycoprotein complexes designated gC-I. J Gen Virol. 1991 Aug;72(Pt 8):1975–1983. doi: 10.1099/0022-1317-72-8-1975. [DOI] [PubMed] [Google Scholar]
  20. Kari B., Liu Y. N., Goertz R., Lussenhop N., Stinski M. F., Gehrz R. Structure and composition of a family of human cytomegalovirus glycoprotein complexes designated gC-I (gB). J Gen Virol. 1990 Nov;71(Pt 11):2673–2680. doi: 10.1099/0022-1317-71-11-2673. [DOI] [PubMed] [Google Scholar]
  21. Knapp S., Bröker M., Amann E. pSEM vectors: high level expression of antigenic determinants and protein domains. Biotechniques. 1990 Mar;8(3):280–281. [PubMed] [Google Scholar]
  22. Kniess N., Mach M., Fay J., Britt W. J. Distribution of linear antigenic sites on glycoprotein gp55 of human cytomegalovirus. J Virol. 1991 Jan;65(1):138–146. doi: 10.1128/jvi.65.1.138-146.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Landini M. P., Re M. C., Mirolo G., Baldassarri B., La Placa M. Human immune response to cytomegalovirus structural polypeptides studied by immunoblotting. J Med Virol. 1985 Dec;17(4):303–311. doi: 10.1002/jmv.1890170403. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Lehner R., Meyer H., Mach M. Identification and characterization of a human cytomegalovirus gene coding for a membrane protein that is conserved among human herpesviruses. J Virol. 1989 Sep;63(9):3792–3800. doi: 10.1128/jvi.63.9.3792-3800.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lussenhop N. O., Goertz R., Wabuke-Bunoti M., Gehrz R., Kari B. Epitope analysis of human cytomegalovirus glycoprotein complexes using murine monoclonal antibodies. Virology. 1988 Jun;164(2):362–372. doi: 10.1016/0042-6822(88)90549-1. [DOI] [PubMed] [Google Scholar]
  27. Matsushita S., Robert-Guroff M., Rusche J., Koito A., Hattori T., Hoshino H., Javaherian K., Takatsuki K., Putney S. Characterization of a human immunodeficiency virus neutralizing monoclonal antibody and mapping of the neutralizing epitope. J Virol. 1988 Jun;62(6):2107–2114. doi: 10.1128/jvi.62.6.2107-2114.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Meyer H., Masuho Y., Mach M. The gp116 of the gp58/116 complex of human cytomegalovirus represents the amino-terminal part of the precursor molecule and contains a neutralizing epitope. J Gen Virol. 1990 Oct;71(Pt 10):2443–2450. doi: 10.1099/0022-1317-71-10-2443. [DOI] [PubMed] [Google Scholar]
  29. Newkirk M. M., Gram H., Heinrich G. F., Ostberg L., Capra J. D., Wasserman R. L. Complete protein sequences of the variable regions of the cloned heavy and light chains of a human anti-cytomegalovirus antibody reveal a striking similarity to human monoclonal rheumatoid factors of the Wa idiotypic family. J Clin Invest. 1988 May;81(5):1511–1518. doi: 10.1172/JCI113483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nowak B., Sullivan C., Sarnow P., Thomas R., Bricout F., Nicolas J. C., Fleckenstein B., Levine A. J. Characterization of monoclonal antibodies and polyclonal immune sera directed against human cytomegalovirus virion proteins. Virology. 1984 Jan 30;132(2):325–338. doi: 10.1016/0042-6822(84)90039-4. [DOI] [PubMed] [Google Scholar]
  31. Pereira L., Hoffman M., Cremer N. Electrophoretic analysis of polypeptides immune precipitated from cytomegalovirus-infected cell extracts by human sera. Infect Immun. 1982 Jun;36(3):933–942. doi: 10.1128/iai.36.3.933-942.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Plotkin S. A., Starr S. E., Friedman H. M., Gonczol E., Brayman K. Vaccines for the prevention of human cytomegalovirus infection. Rev Infect Dis. 1990 Sep-Oct;12 (Suppl 7):S827–S838. doi: 10.1093/clinids/12.supplement_7.s827. [DOI] [PubMed] [Google Scholar]
  33. Silvestri M., Sundqvist V. A., Rudén U., Wahren B. Characterization of a major antigenic region on gp55 of human cytomegalovirus. J Gen Virol. 1991 Dec;72(Pt 12):3017–3023. doi: 10.1099/0022-1317-72-12-3017. [DOI] [PubMed] [Google Scholar]
  34. Skinner M. A., Ting R., Langlois A. J., Weinhold K. J., Lyerly H. K., Javaherian K., Matthews T. J. Characteristics of a neutralizing monoclonal antibody to the HIV envelope glycoprotein. AIDS Res Hum Retroviruses. 1988 Jun;4(3):187–197. doi: 10.1089/aid.1988.4.187. [DOI] [PubMed] [Google Scholar]
  35. Spaete R. R., Thayer R. M., Probert W. S., Masiarz F. R., Chamberlain S. H., Rasmussen L., Merigan T. C., Pachl C. Human cytomegalovirus strain Towne glycoprotein B is processed by proteolytic cleavage. Virology. 1988 Nov;167(1):207–225. doi: 10.1016/0042-6822(88)90071-2. [DOI] [PubMed] [Google Scholar]
  36. Spindler K. R., Rosser D. S., Berk A. J. Analysis of adenovirus transforming proteins from early regions 1A and 1B with antisera to inducible fusion antigens produced in Escherichia coli. J Virol. 1984 Jan;49(1):132–141. doi: 10.1128/jvi.49.1.132-141.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sällberg M., Rudén U., Magnius L. O., Norrby E., Wahren B. Rapid "tea-bag" peptide synthesis using 9-fluorenylmethoxycarbonyl (Fmoc) protected amino acids applied for antigenic mapping of viral proteins. Immunol Lett. 1991 Sep;30(1):59–68. doi: 10.1016/0165-2478(91)90090-w. [DOI] [PubMed] [Google Scholar]
  38. Utz U., Britt W., Vugler L., Mach M. Identification of a neutralizing epitope on glycoprotein gp58 of human cytomegalovirus. J Virol. 1989 May;63(5):1995–2001. doi: 10.1128/jvi.63.5.1995-2001.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Zaia J. A., Forman S. J., Ting Y. P., Vanderwal-Urbina E., Blume K. G. Polypeptide-specific antibody response to human cytomegalovirus after infection in bone marrow transplant recipients. J Infect Dis. 1986 Apr;153(4):780–787. doi: 10.1093/infdis/153.4.780. [DOI] [PubMed] [Google Scholar]

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