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

Abstract

Cell surface expression of the human cytomegalovirus (HCMV) major envelope glycoprotein complex, gp55-116 (gB), was studied by using monoclonal antibodies and an HCMV gp55-116 (gB) recombinant vaccinia virus. HCMV-infected human fibroblasts and recombinant vaccinia virus-infected HeLa cells expressed three electrophoretically distinct proteins of Mr 170,000, 116,000, and 55,000 on their surface. These species have been previously identified within infected cells and purified virions. Two unique neutralizing epitopes were shown to be present on the cell surface gp55-116 (gB). Utilizing HeLa cells infected with the gp55-116 recombinant vaccinia virus as a specific immunosorbent, we have shown that approximately 40 to 70% of the total serum virus-neutralizing activity of a group of individuals with past HCMV infections was directed against this single envelope glycoprotein. The implications of this finding for vaccine development are discussed.

DNA vaccine prime followed by boost with live attenuated virus significantly improves antigen-specific T cell responses against human cytomegalovirus

“…The selection of gB and gM proteins as antigens was based on previous studies showing them as the main targets for neutralizing antibodies. 29,32,33 Due to the small amount of mouse sera, we did not perform a microneutralization assay and instead, used the levels of binding antibodies to reflect the effectiveness of primeboost immunizations. For gB antibody responses, recombinant gB ( Fig.…”

Herpesviral Fcγ receptors: culprits attenuating antiviral IgG?

“…If a prototypic IgG-Fc domain fulfilling the requirements of selective interaction is found, this module could be fused to a large number of HCMV-reactive IgG-Fab domains. Such a cocktail of monoclonal recombinant Abs will be necessary to deal with the interstrain diversity of the major HCMV neutralising epitopes present on the glycoproteins gB and gH [119][120][121] in order to generate widely effective protection. The MCMV animal model could serve as an optimal test tube to generate experimental evidence in vivo for an IgG-Fc-dependent subversive role of vFcgRs in viral pathogenesis in a natural host.…”

Different antibody response to a neutralizing epitope of human cytomegalovirus glycoprotein B among seropositive individuals

“…Secondly, the C23reactive epitope may be less immunogenic in some individuals. A large proportion (40-70%) of the neutralizing antibody response to natural HCMV infection appears to be directed against gB [Britt et al, 1990;Marshall et al, 19921, but Kniess et al [1991] reported that most neutralizing antibodies are directed against the gB epitope AD-1, which is located around amino acid 616 in gB116. However, we did not test reactivity with AD-1 and comparative studies are needed.…”