Identification of Serotype-Specific and Nonserotype-Specific B-Cell Epitopes of Coxsackie B Virus Using Synthetic Peptides

Haarmann, Cäcilia; Schwimmbeck, Peter L.; Mertens, Th.; Schultheiss, Heinz‐Peter; Strauer, B. E.

doi:10.1006/viro.1994.1202

Cited by 37 publications

(23 citation statements)

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“…When the PV type 1 is attached to the cell surface, the amino terminus of subsided VP1 is exposed and may be available for the host immune recognition. Similar conformational changes are observed in non-polio enteroviruses when these viruses infect humans (4,8,10). Furthermore, VP1 of PV is antigenic to B cells and CD4 ϩ T cells and induces neutralizing antibodies in mice and rabbits (7,19).…”

Section: Discussionmentioning

confidence: 62%

Identification of Enteroviruses by Using Monoclonal Antibodies against a Putative Common Epitope

et al. 2003

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A common epitope region of enteroviruses was identified by sequence-independent single-primer amplification (SISPA), followed by immunoscreening of 11 cDNA libraries from two Korean enterovirus isolates (echoviruses 7 and 30) and a coxsackievirus B3 (ATCC-VR 30). The putative common epitope region was localized in the N terminus of VP1 when the displayed recombinant proteins from the phages were chased by the convalescent-phase sera. The genomic region encoding the common epitope region was amplified and then expressed by using the vector pGEX-5X-1. The antigenicity of the expressed recombinant protein was identified by Western blotting with guinea pig antisera for six different serotypes of enteroviruses. After successive immunization of mice with the recombinant common epitope protein, splenocytes were extracted and hybridized with P3X63-Ag8-653 cells. A total of 24 hybridomas that produced monoclonal antibodies (MAbs) against the putative common epitope of enteroviruses were selected. Four of these were immunoglobulin G1 isotypes with a kappa light chain. These MAbs recognized 15 Korean endemic serotypes and prototypes of enteroviruses in an indirect immunofluorescence assay. These results suggest that the expressed protein might be a useful antigen for producing group common antibodies and that the use of the MAbs against the putative common epitope of enteroviruses might be a valuable diagnostic tool for rapidly identifying a broad range of enteroviruses.The genus Enterovirus, belonging to the family Picornaviridae, consists of 66 different subtypes, including polioviruses (PVs), coxsackievirus group A (CVA) and CVB, echoviruses, and the numbered enteroviruses. These viruses cause a wide variety of clinical illnesses, ranging from asymptomatic to fatal, including poliomyelitis, aseptic meningitis, myopericarditis, and respiratory, hepatic, and gastrointestinal infections (15,23,24). Thus, the early diagnosis of enterovirus-related infection from various clinical isolates is important for prognostic, therapeutic, and epidemiologic purposes (1,14,18,26).The primary structure and genetic organization of enteroviruses have been demonstrated recently. Conserved sequences are located in structural and nonstructural proteins of enteroviruses (12,28,29,31,35,39). Therefore, it may be possible to use the conserved region as a common epitope for the serological diagnosis of enteroviruses. The synthetic peptides for the enterovirus group-common epitope could be used for the serological diagnosis of infections caused by a broad range of enteroviruses (3, 25). However, these approaches have not yet been proven effective for the routine laboratory diagnosis of enteroviral antigens in clinical specimens. Thus, it would be useful to develop a rapid and reliable antigen diagnostic method for enteroviral infection by using a monoclonal antibody (MAb) specific for the common epitope region.In the present study, the genetic region of the putative common epitope of enteroviruses was determined by sequenceindependent single-prim...

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Section: Discussionmentioning

confidence: 62%

Identification of Enteroviruses by Using Monoclonal Antibodies against a Putative Common Epitope

et al. 2003

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“…Previous studies have shown that three synthetic peptides based on the sequence of VP1 of CVB3 can identify the serotype-specific and non-serotype-specific Bcell epitopes of coxsackieviruses (7). In that study, three peptides were characterized, showing that peptide VP1-1, representing residues 1-15 of VP1, is highly type-specific for CVB3; peptide VP1-3 (residues 21-35 of VP1) is a non-type-specific antigenic determinant of CVB3; and peptide VP1-24 (residues 229-243 of VP1) is broadly cross-reactive with all coxsackievirus-derived sera, except anti-CVB1 sera.…”

Section: Discussionmentioning

confidence: 99%

“…Moreover, VP2 has an antigenic neutralizing region, termed the "puff region," which is present on the exterior of the virus capsid and is one of the protein regions in poliovirus that line the rim of the canyon floor (6,9,12,15). However, it is principally accepted that, among the four capsid proteins, VP1 has the major antigenic determinants (7).…”

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confidence: 99%

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Production of Cross‐Reactive Peptide Antibodies against Viral Capsid Proteins of Human Enterovirus B to Apply Diagnostic Reagent

Jeong

Ahn

Cho

et al. 2007

Microbiology and Immunology

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The coxsackievirus group B (CVB) of the genus Enterovirus and the species human enterovirus B is a nonenveloped virus containing a single‐stranded positive‐sense RNA genome. Coxsackievirus has icosahedral symmetry and four capsid proteins, VP1, VP2, VP3, and VP4. Specific antibodies against each viral protein are prerequisites for various studies. In this study, we developed seven peptide‐derived antibodies directed against coxsackievirus VP1 (NO1‐NO5), VP2 (B3), and VP3 (GL3). We developed a type‐specific antibody (NO1) and broadly cross‐reactive antibodies (NO3 and NO5) to VP1. Anti‐VP2 and anti‐VP3 antibodies (B3 and GL3, respectively) are also cross‐reactive to human enterovirus B such as CVB and echoviruses. Their sensitivities and reactivities are likely to be better than those of the commercial VP1 monoclonal antibody (MAb). The dot‐blot analysis also showed that NO5 against VP1 is able to detect less than 1 μg [2× 106 plaque‐forming unit (pfu) of CVB3] of viruses, suggesting that it could be used to develop a diagnostic kit that can directly detect human enterovirus B. The antibodies produced here may allow us to undertake several studies, such as those involving viral trafficking, expression kinetics, and the roles of viral proteins in infection, and the development of diagnostic kits.

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“…The P1 region encodes four structural proteins: VP1, VP2, VP3, and VP4. VP1 is the major antigenic determinant of CVB3 and is the most external and immunodominant of its capsid proteins (1). CVB3 can cause meningoencephalitis (2), acute pancreatitis (3), and childhood-onset diabetes (4,5).…”

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confidence: 99%

Development of an Enzyme-Linked Immunosorbent Spot Assay To Measure Serum-Neutralizing Antibodies against Coxsackievirus B3

Yang

Tang

et al. 2014

Clin. Vaccine Immunol.

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Coxsackievirus B3 (CVB3) is the most common pathogen that induces acute and chronic viral myocarditis in children. The cytopathic effect (CPE)-based neutralization test (Nt-CPE) and the plaque reduction neutralization test (PRNT) are the most common methods for measuring neutralizing antibody titers against CVB3 in blood serum samples. However, these two methods are inefficient for CVB3 vaccine clinical trials, which require the testing of a large number of serum specimens. In this study, we developed an efficient neutralization test based on the enzyme-linked immunospot (Nt-ELISPOT) assay for measuring CVB3-neutralizing antibodies. This modified ELISPOT assay was based on the use of a monoclonal antibody against the viral capsid protein VP1 to detect the cells that are infected with CVB3, which, after immunoperoxidase staining, are counted as spots using an automated ELISPOT analyzer. Using the modified ELISPOT assay, we characterized the infection kinetics of CVB3 and divided the infection process of CVB3 on a cluster of cells into four phases. The stability of the Nt-ELISPOT was then evaluated. We found that over a wide range of infectious doses (10 2 to 10 6.5 ؋ 50% tissue culture infectious dose [TCID 50 ] per well), the neutralizing titers of the sera were steady as long as they were tested during the log phase or the first half of the stationary phase of growth of the spots. We successfully shortened the testing period from 7 days to approximately 20 h. We also found that there was a good correlation (R 2 ‫؍‬ 0.9462) between the Nt-ELISPOT and the Nt-CPE assays. Overall, the Nt-ELISPOT assay is a reliable and efficient method for measuring neutralizing antibodies in serum.

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Identification of Serotype-Specific and Nonserotype-Specific B-Cell Epitopes of Coxsackie B Virus Using Synthetic Peptides

Cited by 37 publications

References 0 publications

Identification of Enteroviruses by Using Monoclonal Antibodies against a Putative Common Epitope

Identification of Enteroviruses by Using Monoclonal Antibodies against a Putative Common Epitope

Production of Cross‐Reactive Peptide Antibodies against Viral Capsid Proteins of Human Enterovirus B to Apply Diagnostic Reagent

Development of an Enzyme-Linked Immunosorbent Spot Assay To Measure Serum-Neutralizing Antibodies against Coxsackievirus B3

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