Yeast-produced recombinant virus-like particles of coxsackievirus A6 elicited protective antibodies in mice

Zhou, Yu; Shen, Chaoyun; Zhang, Chao; Zhang, Wei; Wang, Lili; Lan, Ke; Li, Renjie; Huang, Zhong

doi:10.1016/j.antiviral.2016.06.004

Cited by 30 publications

(19 citation statements)

References 40 publications

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“…Mapping of CVA6-specific linear B-cell epitopes within the VP1 protein. CVA6 VLPs are able to potently elicit protective antibodies in mice (19,20). To identify linear B-cell epitopes, a panel of 59 synthetic peptides (designated P1 to P59) covering the entire VP1 region of CVA6 was tested for reaction with the anti-CVA6 VLP mouse serum by ELISA.…”

Section: Figmentioning

confidence: 99%

“…Indeed, efforts have been made to develop CVA6 vaccines using the traditional inactivated whole-virus vaccine approach (15)(16)(17)(18). Recently, our group produced recombinant virus-like particles (VLPs) of CVA6 in insect cells and yeast and evaluated their vaccine potential in preclinical tests (19,20). The CVA6 VLPs efficiently induced serum antibodies that protected mice against lethal viral challenges (19,20), indicating that CVA6 VLPs represent an excellent CVA6 vaccine candidate.…”

mentioning

confidence: 99%

“…Recently, our group produced recombinant virus-like particles (VLPs) of CVA6 in insect cells and yeast and evaluated their vaccine potential in preclinical tests (19,20). The CVA6 VLPs efficiently induced serum antibodies that protected mice against lethal viral challenges (19,20), indicating that CVA6 VLPs represent an excellent CVA6 vaccine candidate. In the present study, we determined for the first time the atomic-resolution (3.0-Å) structure of CVA6 VLPs by cryo-electron microscopy (cryo-EM) single-particle analysis.…”

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confidence: 99%

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A 3.0-Angstrom Resolution Cryo-Electron Microscopy Structure and Antigenic Sites of Coxsackievirus A6-Like Particles

Chen

Zhang

Zhou

et al. 2018

J Virol

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Coxsackievirus A6 (CVA6) has recently emerged as one of the predominant causative agents of hand, foot, and mouth disease (HFMD). The structure of the CVA6 mature viral particle has not been solved thus far. Our previous work shows that recombinant virus-like particles (VLPs) of CVA6 represent a promising CVA6 vaccine candidate. Here, we report the first cryo-electron microscopy (cryo-EM) structure of the CVA6 VLP at 3.0-Å resolution. The CVA6 VLP exhibits the characteristic features of enteroviruses but presents an open channel at the 2-fold axis and an empty, collapsed VP1 pocket, which is broadly similar to the structures of the enterovirus 71 (EV71) VLP and coxsackievirus A16 (CVA16) 135S expanded particle, indicating that the CVA6 VLP is in an expanded conformation. Structural comparisons reveal that two common salt bridges within protomers are maintained in the CVA6 VLP and other viruses of the genus, implying that these salt bridges may play a critical role in enteroviral protomer assembly. However, there are apparent structural differences among the CVA6 VLP, EV71 VLP, and CVA16 135S particle in the surface-exposed loops and C termini of subunit proteins, which are often antigenic sites for enteroviruses. By immunological assays, we identified two CVA6-specific linear B-cell epitopes (designated P42 and P59) located at the GH loop and the C-terminal region of VP1, respectively, in agreement with the structure-based prediction of antigenic sites. Our findings elucidate the structural basis and important antigenic sites of the CVA6 VLP as a strong vaccine candidate and also provide insight into enteroviral protomer assembly. Coxsackievirus A6 (CVA6) is becoming one of the major pathogens causing hand, foot, and mouth disease (HFMD), leading to significant morbidity and mortality in children and adults. However, no vaccine is currently available to prevent CVA6 infection. Our previous work shows that recombinant virus-like particles (VLPs) of CVA6 are a promising CVA6 vaccine candidate. Here, we present a 3.0-Å structure of the CVA6 VLP determined by cryo-electron microscopy. The overall architecture of the CVA6 VLP is similar to those of the expanded structures of enterovirus 71 (EV71) and coxsackievirus A16 (CVA16), but careful structural comparisons reveal significant differences in the surface-exposed loops and C termini of each capsid protein of these particles. In addition, we identified two CVA6-specific linear B-cell epitopes and mapped them to the GH loop and the C-terminal region of VP1, respectively. Collectively, our findings provide a structural basis and important antigenic information for CVA6 VLP vaccine development.

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Section: Figmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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A 3.0-Angstrom Resolution Cryo-Electron Microscopy Structure and Antigenic Sites of Coxsackievirus A6-Like Particles

Chen

Zhang

Zhou

et al. 2018

J Virol

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“…There are two different types of expression strategies for expression plasmids. One is to use two separate expression plasmids, one for producing the P1 region and the other for the 3CD protease (Zhao et al 2013a), and the other strategy is to use a single expression plasmid for both regions (Zhang et al 2015;Zhou et al 2016). In both cases, the VLPs produced had potential as vaccines but Zhang et al obtained significantly higher levels of expression of enterovirus 71 antigens from cells transformed with the single expression plasmid than from those transformed with the two separate plasmids.…”

Section: Multilayered Vlps Produced In Yeastsmentioning

confidence: 99%

Yeast as an expression system for producing virus-like particles: what factors do we need to consider?

Kim

2016

Lett Appl Microbiol

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The development of various types of virus-like particles (VLPs) has accelerated over the past two decades as the importance of VLPs for generating nextgeneration vaccines has been appreciated. Yeast has advantages such as scalable fermentation, low risk of contamination by adventitious agents, low production costs and the ability to produce VLPs with reliable qualities. It is generally recognized that yeast is suitable for producing VLPs that have simple structures and are produced intracellularly. However, recently there has been much effort to extend its applicability, and there is now evidence that it can be used as an expression platform for the productions of VLPs not only of nonenveloped viruses but also of enveloped viruses. Moreover, evidences indicated that yeast allows secretory VLP productions. Meanwhile, it has become evident that the quality and quantity of yeast-derived VLPs are influenced by the choice of plasmid and promoter, the ratio of the structural proteins produced. Here, we review the characteristics of the yeast expression system in terms of the production of VLP and compare it with other expression systems. We also consider strategies for VLP production in yeast and factors that need to be taken into account.

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“…10,11 Both children and adults can be infected by CV-A6, 12,13 patients suffer either from classical HFMD symptoms or atypical HFMD symptoms, such as herpangina, erythema, onychomadesis and epididymitis. 6,14 In order to strengthen the prevention and control of HFMD related diseases, studies on CV-A6 virus-like particle vaccines 15,16 and the establishment of neonatal mouse models have been reported. 17,18 Neutralizing antibody (NtAb) is often used as a major indicator of protective immunity elicited by vaccines and previous infection in epidemiological studies.- 19,20 The cytopathic effect (CPE) is a "gold standard" method to detect NtAb against enteroviruses.…”

Section: Introductionmentioning

confidence: 99%

A surrogate assay for measuring Coxsackievirus A6 neutralizing antibodies

Chen

Gao

et al. 2018

Human Vaccines & Immunotherapeutics

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Coxsackievirus A6 (CV-A6) is one of pathogens causing hand, foot and mouth disease (HFMD) and becomes a new challenge to HFMD control. In this study, we first built a single-round pseudovirus infection system for CV-A6, and then developed a pseudovirus luciferase assay (PVLA) for anti-CV-A6 neutralizing antibody (NtAb) quantification. Since cytopahtic effect (CPE) is considered as the gold standard test for anti-enterovirus NtAb detection, a comparison study has been performed using 318 clinical serum samples, as measured both by PVLA and CPE. The sensitivity and specificity of PVLA was 94.9% (95% CI between 90.8-97.5%) and 92.7% (95% CI between 86.6-96.6%), respectively. Statistical analysis revealed that PVLA and CPE were highly correlated (spearman r = 0.931, P < 0.0001) and in good agreement (94.0%, 95% CI between 90.8-96.4%), showing that PVLA could be used as a surrogate assay for anti-CV-A6 NtAb detection and served as a valuable tool for CV-A6 vaccine evaluation and CV-A6 epidemiological surveillance.

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Yeast-produced recombinant virus-like particles of coxsackievirus A6 elicited protective antibodies in mice

Cited by 30 publications

References 40 publications

A 3.0-Angstrom Resolution Cryo-Electron Microscopy Structure and Antigenic Sites of Coxsackievirus A6-Like Particles

A 3.0-Angstrom Resolution Cryo-Electron Microscopy Structure and Antigenic Sites of Coxsackievirus A6-Like Particles

Yeast as an expression system for producing virus-like particles: what factors do we need to consider?

A surrogate assay for measuring Coxsackievirus A6 neutralizing antibodies

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