Fine Mapping of Neutralization Epitopes on Duck Hepatitis B Virus (DHBV) pre-S Protein Using Monoclonal Antibodies and Overlapping Peptides

Chassot, Sylvie; Lambert, Véronique; Kay, Alan; Godinot, Catherine; Roux, B.; Trépo, C.; Cova, Lucyna

doi:10.1006/viro.1993.1024

Cited by 28 publications

(28 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The major DHBV neutralization epitopes map within the pre-S region of the DHBV large envelope protein is thought to mediate virus-hepatocytes interaction. Binding sites for pre-Sneutralizing antibodies are located in residues 58-66, 83-90, 91-99, 100-107, and 139-145 [21,22]. Our previous researches indicated that a B-cell epitope of residue 127-138 in the envelope protein of DHBV was high antigenic and immunogenic while cross-link it to a protein carrier [23][24][25][26].…”

Section: Discussionmentioning

confidence: 99%

“…C 18 L(AAT TTT TGC CTT CTG ACT TCT TTC CTT CTA TTG) and C 18 D(AAT TCA ATA GAA GGA AAG AAG TCA GAA GGC AAA) encodes for superior HBV CTL epitope C [18][19][20][21][22][23][24][25][26][27] with EcoRI site flanking. The synthetic complementary oligonucleotides of B-cell epitope gene from DHBV pre-S (P 127-138 ) containing flanking EcoR1 restriction sites is as following: b-1: 5 -AAT TGG GGG ACG ATC CAC TCC TGG GAA ATC AGT CTC TCC TCG-3 and b-2: 5 -AAT TCG AGG AGA GAC TGA TTT CCC AGG AGT GGA TCG TCC CCG-3 .…”

Section: Construction Of Epitope-based Dna Vaccinementioning

confidence: 99%

See 1 more Smart Citation

Single B or T-cell epitope-based DNA vaccine using modified vector induces specific immune response against hepadnavirus

Chu

et al. 2005

Immunology Letters

View full text Add to dashboard Cite

Section: Discussionmentioning

confidence: 99%

Section: Construction Of Epitope-based Dna Vaccinementioning

confidence: 99%

Single B or T-cell epitope-based DNA vaccine using modified vector induces specific immune response against hepadnavirus

Chu

et al. 2005

Immunology Letters

View full text Add to dashboard Cite

“…Circumstantial evidence supports a role for DGD in DHBV infection. First, the DGD binding site is highly conserved among the diverse DHBV strains and corresponds to a known neutralizing epitope (5). Second, DGD expression is restricted to DHBV-infectible duck tissues such as the liver, pancreas, and kidney (18,19), although liver-specific expression of transcription factors required for production of viral pregenomic RNA could also restrict hepadnaviral infection to the liver (30).…”

Section: Discussionmentioning

confidence: 99%

Glycine Decarboxylase Mediates a Postbinding Step in Duck Hepatitis B Virus Infection

Tong

Lee

et al. 2004

J Virol

View full text Add to dashboard Cite

The early events in the virus infectious cycle may serve as critical targets for therapeutic intervention. These events are difficult to study due to the transient and dynamic nature of the entry process, the low signal intensity, and the lack of a systemic approach to identify the cellular components required for individual steps. Despite such difficulties, receptors for many important viral pathogens have been identified and general themes in viral attachment and entry are beginning to emerge. For instance, the envelope proteins of orthomyxoviruses, paramyxoviruses, and retroviruses are translated as single polypeptides that are subsequently processed by a basic endopeptidase to generate two polypeptides, one for receptor binding and the other for membrane fusion (13,22). The fusion activity is not triggered until following receptor-ligand interaction and may occur at the site of virus binding (retroviruses) or in the acidic environment of the endosome following virus endocytosis (influenza virus). Furin and PC7 are the known endopeptidases capable of processing gp160 of human immunodeficiency virus (HIV) into the N-terminal gp120 (surface protein) and C-terminal gp41 (transmembrane protein) in the secretory pathway (9). The gp120 remains associated with virus particles via its noncovalent interaction with gp41. Binding of gp120 to CD4, the docking receptor, is accompanied by gp41 interaction with CXCR4 or CCR5, the viral coreceptor, thus leading to fusion of the viral membrane with the plasma membrane of the target cell (1, 6).Such a cleavage event has not been documented for hepatitis B virus (HBV) or its related animal viruses, a group of enveloped hepatotropic DNA viruses (Hepadnaviridae) that, similar to retroviruses, requires reverse transcription for genome replication (7). Instead, HBV expresses three coterminal envelope proteins via alternative translational initiation. Thus, the abundant small envelope protein contains the S domain only, while the large and middle envelope proteins contain additional pre-S1/pre-S2 domains and pre-S2 domain, respectively. The cellular receptor for HBV remains enigmatic following decades of extensive search, as no binding protein for the pre-S1 domain, the candidate site for receptor engagement, turned out to confer HBV susceptibility in reconstitution experiments. Due to the difficulty in obtaining primary human hepatocytes, which may have variable susceptibility to HBV infection, it is not clear whether antibodies against some of the pre-S1 binding partners could block HBV infection. In this regard, duck hepatitis B virus (DHBV) is an ideal model to study the early events in the hepadnaviral life cycle. DHBV can infect ducklings in vivo and primary duck hepatocytes (PDH) in vitro, which remain susceptible to DHBV infection for 2 weeks if maintained in serum-free medium supplemented with dimethyl sulfoxide (DMSO) (25). DHBV expresses two envelope proteins, with a 161-amino-acid pre-S domain unique to the large envelope protein. At least two cellular proteins have been...

show abstract

“…Moreover, in contrast to DHBV envelope protein, for which the humoral response of ducks has been extensively studied and major antigenic regions (ARs) have been precisely mapped by us and others (3,4,28,35), data on the humoral response of ducks to DHBV core protein are scarce. Similarly to HBV, the DHBV core gene encodes two polypeptides: the core protein that forms particles and the secreted e-antigen.…”

mentioning

confidence: 99%

“…To identify DHBc ARs, we used the overlapping peptide scanning technique, an approach initially described by Geysen et al (7,8), which has been successful in the determination of the ARs of different viruses, including the DHBV large envelope protein major antigenic domains (3,4). In the experimentally infected DHBV carrier ducks, anti-DHBc antibodies were demonstrated to be present early after infection, depending of the dose of inoculated virus, and persisted throughout the course of infection, similarly to anti-HBc antibodies in serum of patients with chronic HBV infection (10).…”

mentioning

confidence: 99%

Identification of Antigenic Regions of Duck Hepatitis B Virus Core Protein with Antibodies Elicited by DNA Immunization and Chronic Infection

Thermet

Robaczewska

Rollier

et al. 2004

J Virol

View full text Add to dashboard Cite

The induction of humoral response in ducks by DNA-based immunization against duck hepatitis B virus (DHBV) core protein (DHBc) was investigated. In addition, the amino acid specificity of the induced response was compared by using peptide scanning to that elicited either by protein immunization or during chronic DHBV infection. Immunization of ducks with a plasmid expressing DHBc protein led to the induction of a long-lasting antibody response able to specifically recognize viral protein in chronically infected duck livers. Peptide scanning analysis of anti-DHBc response induced during chronic DHBV infection allowed us to identify six major antigenic regions (AR1 to AR6). The reactivity spectrum of duck sera elicited by protein immunization appeared narrower and was restricted to only four of these antigenic regions in spite of higher anti-DHBc antibody titers. Interestingly, anti-DHBc antibodies induced by DNA-based immunization recognized five of six antigenic regions, and the epitope pattern was broader and more closely related to that observed in chronic viral infections. To gain more insight into the location of antigenic regions, we built a three-dimensional (3-D) model of DHBc protein based on human and duck core sequence alignment data and the HBc 3-D crystal structure. The results suggest that two identified antigenic regions (AR2, amino acids [aa] 64 T-P 84 , and AR5, aa 183 A-R 210 ) are located at positions on the protein surface equivalent to those of the two HBc major epitopes. Moreover, we identified another antigenic region (AR3, aa 99 I-I 112 ) that was recognized by all sera from chronically infected, DNA-or protein-immunized ducks within the large 45-aa insertion in DHBc protein, suggesting that this region, which lacks HBc, is externally exposed.DNA vaccine is a powerful new approach to immunization and immunotherapy that has been shown to induce specific humoral and cellular responses against variety of viral antigens (13). In a genetic vaccine, direct inoculation of plasmid DNA results in the in situ synthesis of antigen by the immunized host; therefore, it has a potential advantage over a recombinant protein immunization in that the resulting antigen would have the conformation and posttranslational modifications identical to those occurring during viral infection (13, 27). Genetic vaccination is also an efficient tool for defining immunogenic epitopes of selected viral antigens. For example, this approach allowed the successful characterization of B-and T-cell epitopes of human immunodeficiency virus Nef, Rev, and Tat proteins (9).The effectiveness of DNA vaccine to Hepatitis B Virus (HBV) envelope proteins has been extensively studied in a murine model and showed induction of high-titer and longlasting humoral responses (19). Studies with a transgenic mouse, lineage E36, which constitutively expresses the HBsAg, strongly suggest the therapeutic efficacy of DNA vaccine encoding viral envelope proteins (17). Moreover, DNA-based vaccination allowed better insight into the murine immune resp...

show abstract

Fine Mapping of Neutralization Epitopes on Duck Hepatitis B Virus (DHBV) pre-S Protein Using Monoclonal Antibodies and Overlapping Peptides

Cited by 28 publications

References 0 publications

Single B or T-cell epitope-based DNA vaccine using modified vector induces specific immune response against hepadnavirus

Single B or T-cell epitope-based DNA vaccine using modified vector induces specific immune response against hepadnavirus

Glycine Decarboxylase Mediates a Postbinding Step in Duck Hepatitis B Virus Infection

Identification of Antigenic Regions of Duck Hepatitis B Virus Core Protein with Antibodies Elicited by DNA Immunization and Chronic Infection

Contact Info

Product

Resources

About