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

Wagner, Beatrice; Kropff, Barbara; Kalbacher, Hubert; Britt, W J; Sundqvist, Vivi‐Anne; Östberg, Lars; Mach, Michael

doi:10.1128/jvi.66.9.5290-5297.1992

Cited by 73 publications

(38 citation statements)

References 39 publications

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“…Construction of the expression plasmid coding for complete gB has been described previously [32]. Carboxyterminal truncated forms of gB were expressed using pcDNA3 as plasmid.…”

Section: Plasmidsmentioning

confidence: 99%

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B Cell Repertoire Analysis Identifies New Antigenic Domains on Glycoprotein B of Human Cytomegalovirus which Are Target of Neutralizing Antibodies

et al. 2011

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Human cytomegalovirus (HCMV), a herpesvirus, is a ubiquitously distributed pathogen that causes severe disease in immunosuppressed patients and infected newborns. Efforts are underway to prepare effective subunit vaccines and therapies including antiviral antibodies. However, current vaccine efforts are hampered by the lack of information on protective immune responses against HCMV. Characterizing the B-cell response in healthy infected individuals could aid in the design of optimal vaccines and therapeutic antibodies. To address this problem, we determined, for the first time, the B-cell repertoire against glycoprotein B (gB) of HCMV in different healthy HCMV seropositive individuals in an unbiased fashion. HCMV gB represents a dominant viral antigenic determinant for induction of neutralizing antibodies during infection and is also a component in several experimental HCMV vaccines currently being tested in humans. Our findings have revealed that the vast majority (>90%) of gB-specific antibodies secreted from B-cell clones do not have virus neutralizing activity. Most neutralizing antibodies were found to bind to epitopes not located within the previously characterized antigenic domains (AD) of gB. To map the target structures of these neutralizing antibodies, we generated a 3D model of HCMV gB and used it to identify surface exposed protein domains. Two protein domains were found to be targeted by the majority of neutralizing antibodies. Domain I, located between amino acids (aa) 133–343 of gB and domain II, a discontinuous domain, built from residues 121–132 and 344–438. Analysis of a larger panel of human sera from HCMV seropositive individuals revealed positivity rates of >50% against domain I and >90% against domain II, respectively. In accordance with previous nomenclature the domains were designated AD-4 (Dom II) and AD-5 (Dom I), respectively. Collectively, these data will contribute to optimal vaccine design and development of antibodies effective in passive immunization.

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“…Construction of the expression plasmid coding for complete gB has been described previously [32]. Carboxyterminal truncated forms of gB were expressed using pcDNA3 as plasmid.…”

Section: Plasmidsmentioning

confidence: 99%

“…Three antigenic domains (AD) have been described previously. AD-1 consists of approximately 80aa between positions 560 and 640 of gB of HCMV strain AD169 [32]. It is the immunodominant region of gB since nearly all sera from HCMV-infected individuals recognize AD-1 [33].…”

Section: Introductionmentioning

confidence: 99%

B Cell Repertoire Analysis Identifies New Antigenic Domains on Glycoprotein B of Human Cytomegalovirus which Are Target of Neutralizing Antibodies

et al. 2011

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“…Recently, data have been reported by Wagner et al [29] suggesting the presence of a conformational epitope in AD1. On the basis of experiments mainly performed with gp58fusion fragments, they reported that the minimal sequwce required for binding of AD1-specific antibodies consists ' of more than 70 amino acids; competition experiments performed with synthetic peptides failed to detect any ability of overlapping 20-amino-acid peptides to inhibit the binding of one neutralizing monoclonal antibody to a gp58-fusion protein.…”

Section: Discussionmentioning

confidence: 84%

“…The presence of a strong human-antibody response, with both HCMV' and EBV' human sera, against the sequence TVDSMIALDI (positions 643 -652) is clearly evident; it has to be emphasized that that this major antibody-binding fragment is not included in the region covered by the set of peptides used in previous studies [29]. Several examples of monoclonal antibodies and animal sera recognizing HSV gB and the corresponding proteins of other herpes viruses have been described [20].…”

Section: Discussionmentioning

confidence: 98%

Characterization of immunoreactive octapeptides of human‐cytomegalovirus gp58

Bonci

Bracci

Caudai

et al. 1993

European Journal of Biochemistry

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We have mapped continuous epitopes, for positions 591 -673 of the human cytomegalovirus 58-kDa glycoprotein using overlapping synthetic peptides and human sera. This region contains a fragment previously described as including the dominant site for induction of human-cytomegalovirus antibodies. Since the selected sequence is highly conserved among herpes viruses, we have considered the possible presence of antigenic cross-reactivity, particularly with the Epstein-Barr virus. Several peptides in the studied region were antigenic and two main continuous epitopes have been identified. Serological cross-reactions observed with Epstein-Barr virus are discussed, focussing on the possible implications of structural features and sequence similarity between humancytomegalovirus and Epstein-Barr-virus glycoproteins.Human-cytomegalovirus (HCMV) infections play an important role as a source of human disease; although primary infection is usually mild or asymptomatic the virus is a major pathogen in immunocompromised individuals and a common cause of congenital defects [l]. A more detailed understanding of the antigenic features involved in the human immune response against HCMV would allow for better control and detection of this infection in susceptible hosts.The HCMV genome codes for 35 -45 structural proteins, most of which are immunogenic [2-41. Outer-envelope glycoproteins, in particular, seem to be major targets for viral neutralization. In several reports, monoclonal antibodies have been described that react with HCMV glycoproteins [5-71, and individually purified or expressed glycoproteins have also been shown to elicit neutralizing responses in experimental animals [8-111. HCMV glycoproteins have been recently grouped into three distinct families, designated gCI-gCIII [ 121. gCI is the major envelope component, consisting of two disulphidelinked protein species of estimated molecular mass 55-58 kDa and 100-116 kDa [13]; these mature forms are generated by proteolytic cleavage of a high-molecular-mass precursor, which is both co-translationally and post-translationally glycosylated [ 14-161.The gene encoding the 58-kDa glycoprotein (gp58) has been mapped and sequenced [ 10, 171 ; the translational product shows sequence similarity with other herpes-virus gly- coproteins, namely the Epstein-Barr-virus (EBV) and herpessimplex-virus (HSV) glycoprotein B (gB), and varicella-herpes-zoster-virus (VHZV) glycoprotein 11, whose role in eliciting a host-immune response has been clearly shown [lo, 17-191. Since cross-reactions of antisera directed against these proteins have been described [20], this feature must be considered for a detailed analysis of HCMV gp58 antigenic properties. These studies are significant, since it has been recently shown that the majority of neutralizing activity in convalescent-human serum is directed against gCI [21], and the isolated complex is also able to induce both humoral and cellular-immune responses in human volunteers [22].In previous studies a HCMV gp58 linear antigenic site, recogniz...

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“…Among these, antigenic domain 1 (AD-1), located around amino acid residue 616, constitutes the major site of recognition by neutralizing antibodies (12). Using fusion proteins expressed in Escherichia coli and synthetic peptides, Wagner et al reported that a continuous sequence of more than 70 amino acids between residues 552 and 630 is essential for antibody binding to AD-1 (13). Furthermore, a strong correlation between the neutralization titer of CMV and antibody levels against AD-1 was observed (5).…”

mentioning

confidence: 99%

Immune Response to Neutralizing Epitope on Human Cytomegalovirus Glycoprotein B in Japanese: Correlation of Serologic Response with HLA‐Type

Wada

Mizuno

Ohta³

et al. 1997

Microbiology and Immunology

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Antigenic domain 1 (AD-1), located between amino acids 608 and 625 of human cytomegalovirus (CMV) gB protein, is the major domain recognized by neutralizing antibodies. Amino acids 552 to 630 are essential for the binding of neutralizing antibodies. We developed an enzyme-linked immunosorbent assay (ELISA) to detect antibodies against a fusion protein containing amino acid residues 549 to 644 of the gB polypeptide and maltose binding protein (MBP). Of 180 seropositive samples, 106 (58.9%) showed positive immuno-reactivity against the fusion protein. None of the seronegative samples reacted with the fusion protein. Among 57 seropositive individuals typed for HLA, subjects with HLA-DR9 had a higher positive rate against the fusion protein (13/14=92.9%) than those without HLA-DR9 (25/43=58.1%). In addition, subjects with HLA-DR15 had a lower positive rate against the fusion protein (7/16=43.3%) than those without HLA-DR15 (31/41=75.6%). Mean OD values of HLA-DR15-positive individuals were significantly lower than those of HLA-DR15-negative individuals. Thus, among CMV-infected individuals, HLA-DR9 may be associated with responders for neutralizing antibodies and HLA-DR15 may be associated with non/lowresponders.

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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

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 * Corresponding author.

Cited by 73 publications

References 39 publications

B Cell Repertoire Analysis Identifies New Antigenic Domains on Glycoprotein B of Human Cytomegalovirus which Are Target of Neutralizing Antibodies

B Cell Repertoire Analysis Identifies New Antigenic Domains on Glycoprotein B of Human Cytomegalovirus which Are Target of Neutralizing Antibodies

Characterization of immunoreactive octapeptides of human‐cytomegalovirus gp58

Immune Response to Neutralizing Epitope on Human Cytomegalovirus Glycoprotein B in Japanese: Correlation of Serologic Response with HLA‐Type

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