Strategies for mapping and imitating viral B-cell epitopes

Cason, John

doi:10.1016/0166-0934(94)90045-0

Cited by 11 publications

(5 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Conformational epitopes consist of several amino acid residues that are discrete in the primary sequence but assemble to form an antigenic determinant on the tertiary structure of the native protein (3,17,23). Conformational epitopes are often formed from more than 10 residues separated from the primary structure (2,4,17). By Western blot analysis, MAb 15F3-1 detected NS1 very effectively in both native and denatured gels (data not shown) and the binding motif for 15F3-1 was located in residues 111 to 116 of the NS1 of DEN-1, indicating that the B-cell epitope of 15F3-1 is a linear epitope.…”

Section: Discussionmentioning

confidence: 99%

Identification of B-Cell Epitope of Dengue Virus Type 1 and Its Application in Diagnosis of Patients

Huang

Chao

et al. 2001

J Clin Microbiol

View full text Add to dashboard Cite

Using a serotype-specific monoclonal antibody (MAb) of dengue virus type 1 (DEN-1), 15F3-1, we identified the B-cell epitope of DEN-1 from a random peptide library displayed on phage. Fourteen immunopositive phage clones that bound specifically to MAb 15F3-1 were selected. These phage-borne peptides had a consensus motif of HxYaWb (a ‫؍‬ S/T, b ‫؍‬ K/H/R) that mimicked the sequence HKYSWK, which corresponded to amino acid residues 111 to 116 of the nonstructural protein 1 (NS1) of DEN-1. Among the four synthetic peptides corresponding to amino acid residues 110 to 117 of the NS1 of DEN-1, -2, -3, and -4, only one peptide, EHKYSWKS (P14M) of DEN-1, was found to bind to 15F3-1 specifically. Furthermore, P14M was shown to inhibit the binding of phage particles to 15F3-1 in a competitive inhibition assay. Histidine 111 (His 111 ) was crucial to the binding of P14M to 15F3-1, since its binding activity dramatically reduced when it changed to leucine 111 (Leu 111 ). This epitope-based peptide demonstrated its clinical diagnostic potential when it reacted with a high degree of specificity with serum samples obtained from both DEN-1-infected rabbits and patients. Based on these observations, our DEN-1 epitope-based serologic test could be useful in laboratory viral diagnosis and in understanding the pathogenesis of DEN-1.Dengue virus (DEN) causes serious febrile illness in humans, including dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) (12). It has been estimated that 100 million cases of dengue fever (DF) occur each year, with a 5% annual case fatality rate even where appropriate supportive treatment is given (10). Primary DEN infection with any of the four serotypes (DEN-1, -2, -3, or -4) often results in a painful, debilitating, but usually nonfatal DF and subsequently leads to protection against reinfection by the same serotype. Many severe and fatal DHF and DSS cases have frequently been reported in regions where more than one serotype of DEN is circulating (9, 11). During epidemics of DHF and DSS, rapid diagnosis of the serotype(s) in patients infected with DEN is important, especially for those patients who visit physicians during the late phase of infection or those patients for whom only convalescent-phase serum samples are available. At present it is still not clear whether DHF and DSS are due to a primary or secondary DEN infection or to other immunopathologic mechanisms (9, 11). Therefore, the identification of B-cell epitopes for DEN can provide important information for the development of a safe and effective dengue vaccine and contribute to the understanding of the pathogenesis of DEN and immunological responses in DEN infection.The B-cell epitopes of DEN-2 had been documented using overlapping synthetic peptides (PEPSCAN) to analyze the antisera (1, 14, 21), and a variety of antigenic domains of DEN-2 had been studied by antigen fragments using recombinant or enzyme cleavage proteins (18,20,26). Phage display, a selection technique in which a peptide or protein is expressed as a fusion w...

show abstract

Section: Discussionmentioning

confidence: 99%

Identification of B-Cell Epitope of Dengue Virus Type 1 and Its Application in Diagnosis of Patients

Huang

Chao

et al. 2001

J Clin Microbiol

View full text Add to dashboard Cite

show abstract

“…However, a partial restoration of discontinuous epitopes in recombinant proteins during Western blotting may also contribute to their reactivity with MAbs (Harper et al, 1990). The incomplete expression of these epitopes in E. coli-expressed recombinant proteins was probably due to the lack of authentic higher-order structure, which is inherent to prokaryotic expression systems (Cason, 1994). Also, it cannot be ruled out that these discontinuous epitopes include residues located outside the identified regions of gBc.…”

mentioning

confidence: 93%

“…Previously, we showed that MAbs specific for discontinuous epitopes of gB retained reactivity against viral gB in a Western blot, although at a relatively low level, even after boiling the samples under reducing conditions before SDS-PAGE (Zaripov et al, 1998). Therefore, we expected that E. coli-expressed recombinant fragments, which presumably FDI lack authentic higher-order structure (Cason, 1994), could be used to map both continuous epitopes and the regions involved in the assembly of discontinuous epitopes. Fig.…”

mentioning

confidence: 99%

Distribution of B-cell epitopes on the pseudorabies virus glycoprotein B.

Zaripov

Morenkov

Fodor

et al. 1999

Journal of General Virology

View full text Add to dashboard Cite

In order to map antigenically important regions of glycoprotein B (gB) of pseudorabies virus (PrV), a panel of recombinant fragments of gB expressed in E. coli and truncated fragments of gB generated by cleavage of purified native gB with trypsin and cyanogen bromide was analysed by using 26 monoclonal antibodies directed against gB. Three continuous epitopes were localized in the vicinity of the N terminus of gB, between amino acids (aa) 59 and 126. One continuous epitope mapped between residues 214 and 279. The residues involved in the assembly of eight discontinuous epitopes were located between aa 540 and 734. The constituents of two discontinuous epitopes were harboured in a segment encompassing aa 540-646. The clustering of continuous epitopes at the extreme N terminus of PrV gB and the locations of residues involved in the assembly of discontinuous epitopes of PrV gB are in good agreement with data on epitope locations in gB homologues from other herpesviruses.

show abstract

“…An appropriate identification of immunogenic motifs in antigens is the first fundamental step when designing epitope-based vaccines [ 30 ]. Advances in knowledge on B-cell epitope sequence and structure have been achieved by techniques such as nuclear magnetic resonance (NMR), and/or X-ray crystallography of antigen-antibody three-dimensional structure complexes [ 31 , 32 ], and by epitope mapping using predictive bioinformatic algorithms [ 33 ] and/or peptide library screening by antibody binding assays [ 34 , 35 , 36 ]. On the other hand, T-cell epitopes can be predicted by means of bioinformatic algorithms and experimentally characterized using MHC multimers and lymphoproliferation or ELISPOT assays [ 37 , 38 ].…”

Section: Overview Of Epitope-based Vaccinesmentioning

confidence: 99%

Peptide-Based Vaccines: Foot-and-Mouth Disease Virus, a Paradigm in Animal Health

et al. 2021

View full text Add to dashboard Cite

Vaccines are considered one of the greatest global health achievements, improving the welfare of society by saving lives and substantially reducing the burden of infectious diseases. However, few vaccines are fully effective, for reasons ranging from intrinsic limitations to more contingent shortcomings related, e.g., to cold chain transport, handling and storage. In this context, subunit vaccines where the essential antigenic traits (but not the entire pathogen) are presented in rationally designed fashion have emerged as an attractive alternative to conventional ones. In particular, this includes the option of fully synthetic peptide vaccines able to mimic well-defined B- and T-cell epitopes from the infectious agent and to induce protection against it. Although, in general, linear peptides have been associated to low immunogenicity and partial protection, there are several strategies to address such issues. In this review, we report the progress towards the development of peptide-based vaccines against foot-and-mouth disease (FMD) a highly transmissible, economically devastating animal disease. Starting from preliminary experiments using single linear B-cell epitopes, recent research has led to more complex and successful second-generation vaccines featuring peptide dendrimers containing multiple copies of B- and T-cell epitopes against FMD virus or classical swine fever virus (CSFV). The usefulness of this strategy to prevent other animal and human diseases is discussed.

show abstract

Strategies for mapping and imitating viral B-cell epitopes

Cited by 11 publications

References 40 publications

Identification of B-Cell Epitope of Dengue Virus Type 1 and Its Application in Diagnosis of Patients

Identification of B-Cell Epitope of Dengue Virus Type 1 and Its Application in Diagnosis of Patients

Distribution of B-cell epitopes on the pseudorabies virus glycoprotein B.

Peptide-Based Vaccines: Foot-and-Mouth Disease Virus, a Paradigm in Animal Health

Contact Info

Product

Resources

About