1887

Abstract

Human cytomegalovirus (HCMV) is a common cause of congenital infection leading to birth defects and a leading cause of serious illness in patients with immunodeficiencies. Studies in this laboratory have focused on a molecular analysis of the immune response to glycoprotein B (gB) of HCMV. This protein has been shown to elicit B cell, helper T cell (T), and cytotoxic T cell responses, suggesting that it may be useful as a subunit HCMV vaccine. However, previous studies showed that although peripheral blood mononuclear cells (PBMC) from all HCMV-seropositive donors proliferate in response to stimulation with whole HCMV, not all donors respond to purified recombinant gB. In the present study, PBMC from HCMV-seropositive donors homozygous for HLA-DR were tested for proliferative responses to whole HCMV and to purified gB expressed in vaccinia virus. PBMC from all donors proliferated in response to HCMV, but those from multiple donors expressing the HLA-DR3Dw3 and -DR4Dw4 specificities, and single donors expressing the -DR15Dw2, -DR13Dw19 and -DR14Dw9 specificities, failed to respond to gB. These results suggested a possible HLA-DR association with low proliferative responses to gB. In further studies, PBMC from donors expressing both putative gB-high responder and low responder HLA-DR alleles were stimulated multiple times with gB to generate gB-specific T cell lines. These cells were then tested for proliferative responses to gB presented by irradiated PBMC sharing only one DR allele with the responder cells. Cells from the gB-specific lines proliferated only when antigen was presented in the context of a responder DR allele but not when presented in the context of a low responder DR allele. Analysis of immune sera revealed that those from donors with PBMC proliferative responses always contained antibodies reactive with B cell epitopes on both the N-terminal gp93 and C-terminal gp55 portions of gB. In contrast, many of the sera from donors with low gB-specific proliferative responses had gp55-specific antibodies but lacked antibodies to gp93. These results suggest that immunogenetic differences in T responsiveness to gB may lead to lack of antigen-specific help for antibody responses to gp93 in some cases. The prevalence of these low responder HLA alleles in the population, and the central importance of the T cell response to the generation of antibodies suggest that native gB alone may not be an attractive candidate for an HCMV subunit vaccine.

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1994-02-01
2024-03-28
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References

  1. Alp N. J., Allport T. D., Vanzanten J., Rodgers B., Sissons J. P. G., Borysiewicz L.K-. 1991; Fine specificity of cellular immune responses in humans to human cytomegalovirus immediate-early 1 protein. Journal of Virology 65:4812–4820
    [Google Scholar]
  2. Banks T., Huo B., Kousoulas K., Spaete R., Pachl C., Pereira L. 1989; A major neutralizing domain maps within the carboxy-terminal half of the cleaved cytomegalovirus B glycoprotein. Journal of General Virology 70:979–985
    [Google Scholar]
  3. Borysiewicz L. K., Hickling J. K., Graham S., Sinclair J., Cranage M. P., Smith G. L., Sissons J. G. P. 1988; Human cytomegalovirus-specific cytotoxic T cells: relative frequency of stage-specific CTL recognizing the 72KD immediate early protein and glycoprotein B expressed by recombinant vaccinia viruses. Journal of Experimental Medicine 168:919–931
    [Google Scholar]
  4. Britt W. J. 1984; Neutralizing antibodies detect a disulfide-linked glycoprotein complex within the envelope of human cytomegalovirus. Virology 135:369–378
    [Google Scholar]
  5. Britt W. J., Vugler L., Butfiloski E. J., Stephens E. B. 1990; Cell surface expression of human cytomegalovirus (HCMV) gp55-116 (gB): use of recombinant vaccinia virus-infected cells in analysis of the human neutralizing antibody response. Journal of Virology 64:1079–1085
    [Google Scholar]
  6. Charpentier B., Michelson S., Martin B. 1986; Definition of HCMV specific target antigens recognized by cytotoxic T cells generated in vitro by using an autologous lymphocyte system. Journal of Immunology 137:330–336
    [Google Scholar]
  7. Chou S. 1992; Comparative analysis of sequence variation in gpl 16 and gp55 components of human cytomegalovirus. Virology 188:388–390
    [Google Scholar]
  8. Forman S. J., Zaia J. A., Clark B. R., Wright C. L., Mills B. J., Pottathil R., Racklin B. C., Gallagher M. T., Welte K., Blume K. G. 1985; A 64,000 dalton matrix protein of human cytomegalovirus induces in vitro immune responses similar to those of whole viral antigen. Journal of Immunology 134:3391–3395
    [Google Scholar]
  9. Gehrz R. C. 1991; Human cytomegalovirus. Biological and clinical perspectives. Advances in Pediatrics 38:190–219
    [Google Scholar]
  10. Gehrz R. C., Christianson W. R., Linner K. M., Groth K. E., Balfour H. H. 1980; Cytomegalovirus vaccine: specific humoral and cellular immune responses in human volunteers. Archives of Internal Medicine 140:936–939
    [Google Scholar]
  11. Gehrz R. C., Fuad S., Liu Y. -N. C., Bach F. H. 1987; HLA class II restriction of T helper cell response to cytomegalovirus (CMV). I. Immunogenetic control of restriction. Journal of Immunology 138:3145–3151
    [Google Scholar]
  12. Gehrz R. C., Liu Y. -N. C., Eckhardt J., Klaus A. 1991; Relevance of immune responses to pathogenesis of cytomegalovirus-associated diseases. Transplantation Proceedings 3: Suppl.3 75–84
    [Google Scholar]
  13. Gehrz R. C., Nelson C. M., Kari B. 1992; A combination of human cytomegalovirus (HCMV)-specific murine monoclonal antibodies exhibits synergistic antiviral activity in vitro . Antiviral Research 17:115–131
    [Google Scholar]
  14. Gold D., Ashley R., Handsfield H. H., Verdon M., Leach L., Millo J., Drew L., Corey L. 1988; Immunoblot analysis of the humoral immune response in primary cytomegalovirus infection. Journal of Infectious Diseases 157:319–326
    [Google Scholar]
  15. Gretch D. R., Suter M., Stinski M. F. 1987; The use of biotinylated monoclonal antibodies and streptavidin affinity chromatography to isolate herpes virus hydrophobic proteins and glycoproteins. Analytical Biochemistry 163:270–277
    [Google Scholar]
  16. Gretch D. R., Gehrz R. C., Stinski M. F. 1988; Characterization of a human cytomegalovirus glycoprotein complex (gcI). Journal of General Virology 69:1205–1215
    [Google Scholar]
  17. Hayes K., Alford C., Britt W. 1987; Antibody response to virus-encoded proteins after cytomegalovirus mononucleosis. Journal of Infectious Diseases 156:615–621
    [Google Scholar]
  18. Kari B., Gehrz R. 1990; Analysis of human antibody responses to human cytomegalovirus envelop glycoproteins found in two families of disulfide linked glycoprotein complexes designated gC-I and gC-II. Archives of Virology 114:213–228
    [Google Scholar]
  19. Kari B., Lussenhop N., Goertz R., Wabuke-Bunoti M., Gehrz R. C. 1986; Characterization of monoclonal antibodies reactive to three biochemically distinct human cytomegalovirus glycoprotein complexes. Journal of Virology 60:345–352
    [Google Scholar]
  20. Kari B., Liu Y. -N. C., Goertz R., Lussenhop N., Stinski M. F., Gehrz R. 1990; Structure, and composition of a family of human cytomegalovirus glycoprotein complexes designated gC-I (gB). Journal of General Virology 71:2673–2680
    [Google Scholar]
  21. Landini M. P., Re M. C., Mirolo G., Baldassarri B., Laplaca M. 1985; Human immune response to cytomegalovirus structural polypeptides studied by immunoblotting. Journal of Medical Vi-rology 17:303–311
    [Google Scholar]
  22. Liu Y. -N. C., Kari B., Gehrz R. C. 1988a; Immune responses to major human cytomegalovirus glycoprotein complexes. Journal of Virology 62:1066–1070
    [Google Scholar]
  23. Liu Y. -N. C., Eckhardt J., Kari B., Gehrz R. C. 1988b; General characterization of human cytomegalovirus-specific proliferative CD4+ T cell clones. International Journal of Cell Cloning 2:352–364
    [Google Scholar]
  24. Liu Y. -N. C., Klaus A., Kari B., Stinski M. F., Eckhardt J., Gehrz R. C. 1991a; The N-terminal 513 amino acids of the envelope glycoprotein gB of human cytomegalovirus stimulates both B and T cell immune responses in humans. Journal of Virology 65:1644–1648
    [Google Scholar]
  25. Liu Y. -N. C., Opitz G., Klaus A., Cooper J., Gehrz R. C. 1991b; Helper T cell (Th) epitope mapping of gp93 in gcI complexes of human cytomegalovirus (HCMV). In Progress in Cytomegalovirus Research pp 187–190 Landini M. P. Edited by
    [Google Scholar]
  26. Liu Y. -N. C., Curtsinger J., Donahue P. R., Klaus A., Opitz G., Cooper J., Karr R. W., Bach F. H., Gehrz R. C. 1993; Molecular analysis of the immune response to human cytomegalovirus glycoprotein B. I. Mapping of HLA-restricted helper T cell epitopes on gp93. Journal of General Virology 74:2207–2214
    [Google Scholar]
  27. Lussenhop N. O., Goertz R., Wabuke-Bunoti M., Gehrz R., Kari B. 1988; Epitope analysis of human cytomegalovirus glycoprotein complexes using murine monoclonal antibodies. Virology 164:362–372
    [Google Scholar]
  28. Masuho Y., Matsumoto Y. -I., Sugano T., Fujinaga S., Minamishima Y. 1987; Human monoclonal antibodies neutralizing human cytomegalovirus. Journal of General Virology 68:1457–1461
    [Google Scholar]
  29. Meyer H., Sundqvist V. -A., Pereira L., Mach M. 1992; Glycoprotein gpl 16 of human cytomegalovirus contains epitopes for strain-common and strain-specific antibodies. Journal of General Virology 73:2375–2383
    [Google Scholar]
  30. Ohlin M., Sundqvist V. -A., Mach M., Wahren B., Borrebaeck C. A. K. 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. Journal of Virology 67:703–710
    [Google Scholar]
  31. Pereira L., Hoffman M., Cremer N. 1982a; Electrophoretic analysis of polypeptides immune precipitated from cytomegalovirus-infected cell extracts by human sera. Infection and Immunity 36:933–942
    [Google Scholar]
  32. Pereira L., Hoffman M., Gallo D., Cremer N. 1982b; Monoclonal antibodies to human cytomegalovirus: three surface membrane proteins with unique immunological and electrophoretic properties specify cross-reactive determinants. Infection and Immunity 36:924–932
    [Google Scholar]
  33. Pereira L., Hoffman M., Tatsuno M., Dondero J. 1984; Polymorphism of human cytomegalovirus glycoproteins characterized by monoclonal antibodies. Virology 139:73–86
    [Google Scholar]
  34. Rasmussen L., Mullenax J., Nelson R., Merigan T. C. 1985; Viral polypeptides detected by a complement-dependent neutralizing murine monoclonal antibody to human cytomegalovirus. Journal of Virology 55:274–280
    [Google Scholar]
  35. Rasmussen L. 1990; Immune response to human cytomegalovirus infection. Current Topics in Microbiology and Immunology 154:221–254
    [Google Scholar]
  36. Riddell S. R., Rabin M., Geballe A. P., Britt W. J., Greenberg P. D. 1991; Class I MHC-restricted cytotoxic T lymphocyte recognition of cells infected with human cytomegalovirus does not require endogenous viral gene expression. Journal of Immunology 146:2795–2804
    [Google Scholar]
  37. Rodgers B., Borysiewicz L., Mundin J., Graham S., Sissons P. 1987; Immunoaffinity purification of a 72K early antigen of human cytomegalovirus: analysis of humoral and cell-mediated immunity to the purified polypeptide. Journal of General Virology 68:2371–2378
    [Google Scholar]
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