Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1993 Feb;67(2):703–710. doi: 10.1128/jvi.67.2.703-710.1993

Fine specificity of the human immune response to the major neutralization epitopes expressed on cytomegalovirus gp58/116 (gB), as determined with human monoclonal antibodies.

M Ohlin 1, V A Sundqvist 1, M Mach 1, B Wahren 1, C A Borrebaeck 1
PMCID: PMC237421  PMID: 7678304

Abstract

The humoral immune response to human cytomegalovirus (CMV) membrane glycoprotein gp58/116 (gB) has been studied by establishing cell lines producing specific human monoclonal antibodies. These cell lines were generated from peripheral blood lymphocytes obtained from a healthy carrier. Hybridomas producing gp58/116-specific antibodies were detected by reactivity to procaryotically expressed proteins containing the major neutralizing epitopes of this glycoprotein complex. One antibody, ITC88, which recognized an epitope located between amino acid residues 67 and 86 of gp116, potently neutralized the virus at 1 to 2 micrograms of immunoglobulin G per ml. Only four of the six human antibodies detecting the major neutralizing domain of gp58 neutralized the virus, and none of them required complement for activity. All antibodies that bound mature, processed gp58 recognized a conformational epitope involving sequences between residues 549 and 635. However, small differences existed between the antibodies in the actual minimal requirement for C- and N-terminal parts of this epitope. By peptide mapping with several of the antibodies, the epitope was shown to consist mainly of residues between amino acids 570 to 579 and 606 to 619. Despite the conformational nature of the epitope, the antibodies recognized both reduced and denatured native antigen. Presence of carbohydrates was not required for antigen binding of these gp58-specific human antibodies, but in at least one case, it greatly enhanced antigen recognition, indicating an importance of carbohydrate structures in some epitopes within the major neutralizing specificity of gp58.

Full text

PDF
707

Images in this article

707Image
on p.707
708Image
on p.708

Selected References

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

  1. Ahlfors K. Epidemiological studies of congenital cytomegalovirus infection. Scand J Infect Dis Suppl. 1982;34:1–36. doi: 10.3109/inf.1982.14.suppl-34.01. [DOI] [PubMed] [Google Scholar]
  2. Beatty J. D., Beatty B. G., Vlahos W. G. Measurement of monoclonal antibody affinity by non-competitive enzyme immunoassay. J Immunol Methods. 1987 Jun 26;100(1-2):173–179. doi: 10.1016/0022-1759(87)90187-6. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. 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]
  5. Britt W. J., Vugler L., Stephens E. B. Induction of complement-dependent and -independent neutralizing antibodies by recombinant-derived human cytomegalovirus gp55-116 (gB). J Virol. 1988 Sep;62(9):3309–3318. doi: 10.1128/jvi.62.9.3309-3318.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Carroll W. L., Thielemans K., Dilley J., Levy R. Mouse x human heterohybridomas as fusion partners with human B cell tumors. J Immunol Methods. 1986 May 1;89(1):61–72. doi: 10.1016/0022-1759(86)90032-3. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. 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]
  10. Gilljam G., Wahren B. Properties of IgG subclasses to human cytomegalovirus. J Virol Methods. 1989 Aug;25(2):139–151. doi: 10.1016/0166-0934(89)90028-1. [DOI] [PubMed] [Google Scholar]
  11. Grunow R., Jahn S., Porstmann T., Kiessig S. S., Steinkellner H., Steindl F., Mattanovich D., Gürtler L., Deinhardt F., Katinger H. The high efficiency, human B cell immortalizing heteromyeloma CB-F7. Production of human monoclonal antibodies to human immunodeficiency virus. J Immunol Methods. 1988 Feb 10;106(2):257–265. doi: 10.1016/0022-1759(88)90206-2. [DOI] [PubMed] [Google Scholar]
  12. Johnson V. G., Schlom J., Paterson A. J., Bennett J., Magnani J. L., Colcher D. Analysis of a human tumor-associated glycoprotein (TAG-72) identified by monoclonal antibody B72.3. Cancer Res. 1986 Feb;46(2):850–857. [PubMed] [Google Scholar]
  13. Kari B., Gehrz R. Analysis of human antibody responses to human cytomegalovirus envelope glycoproteins found in two families of disulfide linked glycoprotein complexes designated gC-I and gC-II. Arch Virol. 1990;114(3-4):213–228. doi: 10.1007/BF01310750. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Kari B., Lussenhop N., Goertz R., Wabuke-Bunoti M., Radeke R., Gehrz R. Characterization of monoclonal antibodies reactive to several biochemically distinct human cytomegalovirus glycoprotein complexes. J Virol. 1986 Nov;60(2):345–352. doi: 10.1128/jvi.60.2.345-352.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  18. Linde A., Andersson J., Lundgren G., Wahren B. Subclass reactivity to Epstein-Barr virus capsid antigen in primary and reactivated EBV infections. J Med Virol. 1987 Feb;21(2):109–121. doi: 10.1002/jmv.1890210203. [DOI] [PubMed] [Google Scholar]
  19. Linde A., Dahl H., Wahren B., Fridell E., Salahuddin Z., Biberfeld P. IgG antibodies to human herpesvirus-6 in children and adults and in primary Epstein-Barr virus infections and cytomegalovirus infections [corrected]. J Virol Methods. 1988 Sep;21(1-4):117–123. doi: 10.1016/0166-0934(88)90058-4. [DOI] [PubMed] [Google Scholar]
  20. Liu Y. N., Kari B., Gehrz R. C. Human immune responses to major human cytomegalovirus glycoprotein complexes. J Virol. 1988 Mar;62(3):1066–1070. doi: 10.1128/jvi.62.3.1066-1070.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ljungman P., Wahren B., Sundqvist V. A. Lymphocyte proliferation and IgG production with herpesvirus antigens in solid phase. J Virol Methods. 1985 Dec;12(3-4):199–208. doi: 10.1016/0166-0934(85)90130-2. [DOI] [PubMed] [Google Scholar]
  22. Ljungman P., Zetterqvist L., Sundqvist V. A., Jeansson S., Heimdahl A., Wahren B. Lymphocyte and IgG responses to different herpes simplex virus antigens in patients with frequent HSV-1 reactivations. Clin Exp Immunol. 1988 May;72(2):207–210. [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Masuho Y., Matsumoto Y., Sugano T., Fujinaga S., Minamishima Y. Human monoclonal antibodies neutralizing human cytomegalovirus. J Gen Virol. 1987 May;68(Pt 5):1457–1461. doi: 10.1099/0022-1317-68-5-1457. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Meyer H., Sundqvist V. A., Pereira L., Mach M. Glycoprotein gp116 of human cytomegalovirus contains epitopes for strain-common and strain-specific antibodies. J Gen Virol. 1992 Sep;73(Pt 9):2375–2383. doi: 10.1099/0022-1317-73-9-2375. [DOI] [PubMed] [Google Scholar]
  27. Meyers J. D., Leszczynski J., Zaia J. A., Flournoy N., Newton B., Snydman D. R., Wright G. G., Levin M. J., Thomas E. D. Prevention of cytomegalovirus infection by cytomegalovirus immune globulin after marrow transplantation. Ann Intern Med. 1983 Apr;98(4):442–446. doi: 10.7326/0003-4819-98-4-442. [DOI] [PubMed] [Google Scholar]
  28. Ohlin M., Sundqvist V. A., Gilljam G., Rudén U., Gombert F. O., Wahren B., Borrebaeck C. A. Characterization of human monoclonal antibodies directed against the pp65-kD matrix antigen of human cytomegalovirus. Clin Exp Immunol. 1991 Jun;84(3):508–514. [PMC free article] [PubMed] [Google Scholar]
  29. Parry N. R., Barnett P. V., Ouldridge E. J., Rowlands D. J., Brown F. Neutralizing epitopes of type O foot-and-mouth disease virus. II. Mapping three conformational sites with synthetic peptide reagents. J Gen Virol. 1989 Jun;70(Pt 6):1493–1503. doi: 10.1099/0022-1317-70-6-1493. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Radsak K., Brücher K. H., Britt W., Shiou H., Schneider D., Kollert A. Nuclear compartmentation of glycoprotein B of human cytomegalovirus. Virology. 1990 Aug;177(2):515–522. doi: 10.1016/0042-6822(90)90516-t. [DOI] [PubMed] [Google Scholar]
  32. Rasmussen L. E., Nelson R. M., Kelsall D. C., Merigan T. C. Murine monoclonal antibody to a single protein neutralizes the infectivity of human cytomegalovirus. Proc Natl Acad Sci U S A. 1984 Feb;81(3):876–880. doi: 10.1073/pnas.81.3.876. [DOI] [PMC free article] [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. 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]
  35. Sundqvist V. A. Frequency and specificity of varicella zoster virus IgM response. J Virol Methods. 1982 Nov;5(3-4):219–227. doi: 10.1016/0166-0934(82)90012-x. [DOI] [PubMed] [Google Scholar]
  36. 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]
  37. Tomiyama T., Masuho Y. Antiviral activities of a human monoclonal antibody against human cytomegalovirus. Adv Exp Med Biol. 1990;278:105–113. doi: 10.1007/978-1-4684-5853-4_11. [DOI] [PubMed] [Google Scholar]
  38. Tomiyama T., Matsumoto Y., Sugano T., Fujinaga S., Masuho Y. Preparation of human monoclonal antibodies against a cytomegalovirus glycoprotein complex of 130 and 55 kDa. J Immunol Methods. 1990 Aug 7;131(2):249–255. doi: 10.1016/0022-1759(90)90196-3. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. Wagner B., Kropff B., Kalbacher H., Britt W., Sundqvist V. A., Ostberg L., Mach M. 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. J Virol. 1992 Sep;66(9):5290–5297. doi: 10.1128/jvi.66.9.5290-5297.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wahren B., Oberg B. Inhibition of cytomegalovirus late antigens by phosphonoformate. Intervirology. 1980;12(6):335–339. doi: 10.1159/000149093. [DOI] [PubMed] [Google Scholar]
  42. Winston D. J., Ho W. G., Lin C. H., Bartoni K., Budinger M. D., Gale R. P., Champlin R. E. Intravenous immune globulin for prevention of cytomegalovirus infection and interstitial pneumonia after bone marrow transplantation. Ann Intern Med. 1987 Jan;106(1):12–18. doi: 10.7326/0003-4819-106-1-12. [DOI] [PubMed] [Google Scholar]
  43. Woodward M. P., Young W. W., Jr, Bloodgood R. A. Detection of monoclonal antibodies specific for carbohydrate epitopes using periodate oxidation. J Immunol Methods. 1985 Apr 8;78(1):143–153. doi: 10.1016/0022-1759(85)90337-0. [DOI] [PubMed] [Google Scholar]
  44. van der Voort L. H., de Leij L. F., The T. H. Characterization of membrane antigens on human cytomegalovirus-infected fibroblasts recognized by human antibodies. J Virol. 1989 Mar;63(3):1485–1488. doi: 10.1128/jvi.63.3.1485-1488.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES