Neutralizing Antibodies Induced by Recombinant Virus-Like Particles of Enterovirus 71 Genotype C4 Inhibit Infection at Pre- and Post-attachment Steps

Ku, Zhiqiang; Ye, Xiaohua; Huang, Xulin; Cai, Yuepiao; Li, Renjie; Li, Yan; Su, Zhiguo; Huang, Zhong

doi:10.1371/journal.pone.0057601

Cited by 66 publications

(74 citation statements)

References 27 publications

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“…RD and Vero cells were grown as described previously (18). EV71 strains used in the present study included EV71/BrCr, EV71/ G081, EV71/G082, EV71/FY2, EV71/SH98, and a mouse-adapted virus termed EV71/MAV-VR.…”

Section: Cells and Virusesmentioning

confidence: 99%

“…EU262658) has been described previously (19). All viruses were propagated in RD cells as described previously (18,19). The virus titers were determined by the Reed and Muench method based on a typical cytopathic effect (CPE) developing in the infected RD cells and expressed in 50% tissue culture infectious doses (TCID 50 ).…”

Section: Cells and Virusesmentioning

confidence: 99%

“…Ltd. (Shanghai, China). Overlapping peptides, which span the entire amino acid sequence of VP1, have been described previously (18). Each peptide contains 15 amino acid residues.…”

Section: Cells and Virusesmentioning

confidence: 99%

“…The pelleted E-and F-particles were resuspended in PBS and stored at Ϫ80°C until use. Recombinant EV71 VLP was prepared as described previously (18). Recombinant EV71 VP1 protein was described previously (21).…”

Section: Cells and Virusesmentioning

confidence: 99%

“…Protein samples were subjected to SDS-12% PAGE and Western blotting as described previously (18) with the following modifications: the membranes were probed with a mouse anti-HBc monoclonal antibody (catalog no. ab8638; Abcam, Cambridge, MA) or a mouse anti-His monoclonal antibody (Abmart, Shanghai, China), followed by incubation with horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (Sigma).…”

Section: Cells and Virusesmentioning

confidence: 99%

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Chimeric Virus-Like Particle Vaccines Displaying Conserved Enterovirus 71 Epitopes Elicit Protective Neutralizing Antibodies in Mice through Divergent Mechanisms

et al. 2014

J Virol

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Enterovirus 71 (EV71) is a major causative agent of hand, food, and mouth disease, which frequently occurs in young children. Since there are 11 subgenotypes (A, B1 to B5, and C1 to C5) within EV71, an EV71 vaccine capable of protecting against all of these subgenotypes is desirable. We report here the vaccine potential and protective mechanism of two chimeric virus-like particles (VLPs) presenting conserved neutralizing epitopes of EV71. We show that fusions of hepatitis B core antigen (HBc) with the SP55 or SP70 epitope of EV71, designated HBcSP55 and HBcSP70, respectively, can be rapidly generated and self-assembled into VLPs with the epitopes displayed on the surface. Immunization with the chimeric VLPs induced carrier-and epitope-specific antibody responses in mice. Anti-HBcSP55 and anti-HBcSP70 sera, but not anti-HBc sera, were able to neutralize in vitro multiple genotypes and strains of EV71. Importantly, passive immunization with anti-HBcSP55 or anti-HBcSP70 sera protected neonatal mice against lethal EV71 infections. Interestingly, anti-HBcSP70 sera could inhibit EV71 attachment to susceptible cells, whereas anti-HBcSP55 sera could not. However, both antisera were able to neutralize EV71 infection in vitro at the postattachment stage. The divergent mechanism of neutralization and protection conferred by anti-SP70 and anti-SP55 sera is in part attributed to their respective ability to bind authentic viral particles. Collectively, our study not only demonstrates that chimeric VLPs displaying the SP55 and SP70 epitopes are promising candidates for a broad-spectrum EV71 vaccine but also reveals distinct mechanisms of neutralization by the SP55-and SP70-targeted antibodies. E nterovirus 71 (EV71) is the major causative agent of hand, foot, and mouth disease, which is prevalent in the Asia-Pacific region. EV71 infection may result in severe neurological complications and even death (1-3). However, there is currently no available EV71 vaccine (4, 5).EV71 is a member of the enterovirus genus of the Picornaviridae family. It possesses a single-stranded, positive-sense RNA genome, which is encapsidated within an icosahedral capsid consisting of 60 copies of each of VP1, VP2, VP3, and VP4 subunit proteins. Based on the VP1 sequence, EV71 is categorized into three genotypes (A, B, and C), which can be further divided into eleven subgenotypes (A, B1 to B5, and C1 to C5) (reviewed in references 1 and 2). Thus, an EV71 vaccine capable of protecting against all of these subgenotypes is desirable.Increasing evidence has established that neutralizing antibodies play a key role in in vivo protection against lethal EV71 infection (reviewed in references 4 and 5). For example, Foo et al. showed that passive transfer of neutralizing antisera protects recipient mice against lethal EV71 challenge (6). Inactivated wholevirus and recombinant EV71 virus-like particles (VLPs) containing all capsid subunit proteins could elicit potent neutralizing antibodies (reviewed in references 4 and 5). Besides intact capsids, VP1 has been sh...

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Section: Cells and Virusesmentioning

confidence: 99%

Section: Cells and Virusesmentioning

confidence: 99%

“…Ltd. (Shanghai, China). Overlapping peptides, which span the entire amino acid sequence of VP1, have been described previously (18). Each peptide contains 15 amino acid residues.…”

Section: Cells and Virusesmentioning

confidence: 99%

Section: Cells and Virusesmentioning

confidence: 99%

Section: Cells and Virusesmentioning

confidence: 99%

See 3 more Smart Citations

Chimeric Virus-Like Particle Vaccines Displaying Conserved Enterovirus 71 Epitopes Elicit Protective Neutralizing Antibodies in Mice through Divergent Mechanisms

et al. 2014

J Virol

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Virus-like particles in picornavirus vaccine development

Dong

Guo

Sun

2014

Appl Microbiol Biotechnol

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Virus-like particles (VLP), which are similar to natural virus particles but do not contain viral genes, have brought about significant breakthroughs in many research fields because of their unique advantages. The ordered repeating epitopes of VLP can induce immunity responses similar to those prompted by natural viral infection; thus, VLP vaccines are regarded as candidate alternatives to whole-virus vaccines. As picornavirus has serious impacts on human and animal health, the development of efficient and safe vaccines is a key endeavor in preventing virus infections. The characteristics of picornavirus capsid proteins allow the development of VLP vaccines. This paper investigates research scenarios and progress on picornavirus VLP vaccines with the aim of providing a reference for researchers focusing on virology and vaccinology.

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Generation and characterization of a protective mouse monoclonal antibody against human enterovirus 71

Deng

et al. 2015

Appl Microbiol Biotechnol

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Human enterovirus 71 (EV71) infection has emerged as a major threat to children; however, no effective antiviral treatment or vaccine is currently available. Antibody-based treatment shows promises to control this growing public health problem of EV71 infection, and a few potent monoclonal antibodies (mAbs) targeting viral capsid protein have been well described. Here, we generated an EV71-specific mouse mAb 2G8 that conferred full protection against lethal EV71 challenge in a suckling mouse model. 2G8 belonged to IgM isotype and neutralized EV71 at the attachment stage. Biochemical assays mapped the binding epitope of 2G8 to the SP70 peptide, which spanning amino acid residues 208-222 on the VP1 protein. Alanine scanning mutagenesis defined the essential roles of multiple residues, including Y208, T210, G212, K215, K218, L220, E221, and Y222, for 2G8 binding. Then, a panel of single mutation was individually introduced into the EV71 infectious clone by reverse genetics, and three mutant viruses, K215A, K218A, and L220A, were successfully recovered and characterized. Biochemical and neutralization assays revealed that K218A mutant partially escaped 2G8 neutralization, while L220A completely abolished 2G8 binding and neutralization. In particular, neutralization assays with human sera demonstrated that K218A and L220A substitutions are also critical for antibody neutralization in natural infection population. These findings not only generate a protective mAb candidate with therapeutic potential but also provide insights into antibody-mediated EV71 neutralization mechanism.

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Neutralizing Antibodies Induced by Recombinant Virus-Like Particles of Enterovirus 71 Genotype C4 Inhibit Infection at Pre- and Post-attachment Steps

Cited by 66 publications

References 27 publications

Chimeric Virus-Like Particle Vaccines Displaying Conserved Enterovirus 71 Epitopes Elicit Protective Neutralizing Antibodies in Mice through Divergent Mechanisms

Chimeric Virus-Like Particle Vaccines Displaying Conserved Enterovirus 71 Epitopes Elicit Protective Neutralizing Antibodies in Mice through Divergent Mechanisms

Virus-like particles in picornavirus vaccine development

Generation and characterization of a protective mouse monoclonal antibody against human enterovirus 71

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