Expression and immunogenicity of novel subunit enterovirus 71 VP1 antigens

Xu, Juan; Wang, Shixia; Gan, Weihua; Zhang, Wenhong; Ju, Li; Huang, Zhibin; Lu, Shan

doi:10.1016/j.bbrc.2012.03.067

Cited by 7 publications

(12 citation statements)

References 23 publications

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“…1C). VP1, an outer surface capsid protein of EV71, is reportedly involved in virus infectivity (68) and immunogenicity (69,70). VP1 has been shown to interact with the cell surface receptors SCARB2 and PSGL-1 (71,72) and with the binding molecules lactoferrin and annexin IIA (10,73).…”

Section: Figmentioning

confidence: 99%

Cell Surface Nucleolin Facilitates Enterovirus 71 Binding and Infection

Wang

Huang

et al. 2015

J Virol

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Because the pathogenesis of enterovirus 71 (EV71) remains mostly ambiguous, identifying the factors that mediate viral binding and entry to host cells is indispensable to ultimately uncover the mechanisms that underlie virus infection and pathogenesis. Despite the identification of several receptors/attachment molecules for EV71, the binding, entry, and infection mechanisms of EV71 remain unclear. Herein, we employed glycoproteomic approaches to identify human nucleolin as a novel binding receptor for EV71. Glycoproteins purified by lectin chromatography from the membrane extraction of human cells were treated with sialidase, followed by immunoprecipitation with EV71 particles. Among the 16 proteins identified by tandem mass spectrometry analysis, cell surface nucleolin attracted our attention. We found that EV71 interacted directly with nucleolin via the VP1 capsid protein and that an antinucleolin antibody reduced the binding of EV71 to human cells. In addition, the knockdown of cell surface nucleolin decreased EV71 binding, infection, and production in human cells. Furthermore, the expression of human nucleolin on the cell surface of a mouse cell line increased EV71 binding and conferred EV71 infection and production in the cells. These results strongly indicate that human nucleolin can mediate EV71 binding to and infection of cells. Our findings also demonstrate that the use of glycoproteomic approaches is a reliable methodology to discover novel receptors for pathogens. IMPORTANCEOutbreaks of EV71 have been reported in Asia-Pacific countries and have caused thousands of deaths in young children during the last 2 decades. The discovery of new EV71-interacting molecules to understand the infection mechanism has become an emergent issue. Hence, this study uses glycoproteomic approaches to comprehensively investigate the EV71-interacting glycoproteins. Several EV71-interacting glycoproteins are identified, and the role of cell surface nucleolin in mediating the attachment and entry of EV71 is characterized and validated. Our findings not only indicate a novel target for uncovering the EV71 infection mechanism and anti-EV71 drug discovery but also provide a new strategy for virus receptor identification.

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Section: Figmentioning

confidence: 99%

Cell Surface Nucleolin Facilitates Enterovirus 71 Binding and Infection

Wang

Huang

et al. 2015

J Virol

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“…The four structural proteins are the major Ags of EV71 and have thus been the focus of vaccine development and applied as the main effective constituents in vaccine effectiveness evaluations (13,14). In particular, VP1 is highly exposed, as well as considered the main target of neutralizing Abs, and has thus been broadly used for subunit vaccine development (15)(16)(17). The lower prevalence of neutralizing Abs accompanied by higher transmission rates in children and infants indicates that neutralizing Abs are important for the prevention of EV71 infection (8,18).…”

mentioning

confidence: 99%

VP2 Dominated CD4+ T Cell Responses against Enterovirus 71 and Cross-Reactivity against Coxsackievirus A16 and Polioviruses in a Healthy Population

Tan

Sun

et al. 2013

The Journal of Immunology

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Enterovirus 71 (EV71)–associated hand-foot-mouth disease has become a major threat to public health in the Asia–Pacific region. Although T cell immunity is closely correlated with clinical outcomes of EV71 infection, little is known about T cell immunity baseline against EV71 and T cell immunogenecity of EV71 Ags in the population, which has restricted our understanding of immunoprotection mechanisms. In this study, we investigated the cellular immune responses against the four structural Ags of EV71 and determined the immunohierarchy of these Ags in healthy adults. A low frequency of EV71-responsive T cells was detected circulating in peripheral blood, and broad T cell immune responses could be identified in most of the subjects after in vitro expansion. We demonstrated that the VP2 Ag with broad distribution of immunogenic peptides dominates T cell responses against EV71 compared with VP1, VP3, and VP4. Furthermore, the responses were illuminated to be mainly single IFN-γ–secreting CD4+ T cell dependent, indicating the previous natural acute viral infection of the adult population. Conservancy analysis of the immunogenic peptides revealed that moderately variant peptides were in the majority in coxsackievirus A16 (CV-A16) whereas most of the peptides were highly variant in polioviruses. Less efficient cross-reactivity against CV-A16 might broadly exist among individuals, whereas influences derived from poliovirus vaccination would be limited. Our findings suggest that the significance of VP2 Ag should be addressed in the future EV71-responsive immunological investigations. And the findings concerning the less efficient cross-reactivity against CV-A16 and limited influences from poliovirus vaccination in EV71-contacted population would contribute to a better understanding of immunoprotection mechanisms against enteroviruses.

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“…In our previous research, a pilot antigen engineering study was conducted to explore novel VP1 designs to improve expression and immunogenicity of VP1 antigen [ 21 ] . Five codon-optimized VP1 DNA vaccines, including wt-VP1, tPA-VP1, tPA-VP1-dimer (VP1-d), tPA-VP1-hFc (VP1-hFc) and tPA-VP1-mFc (VP1-mFc), were constructed and tested for in vitro expression and in vivo immunogenicity.…”

Section: Introductionmentioning

confidence: 99%

“…In this study, we investigated possible factors that could affect the expression, secretion, and immuno- genicity of VP1 DNA vaccines. In our previous study, we showed that VP1-mFc expressed at significantly lower levels and failed to mount an effective immune response [ 21 ] . In subsequent experiments, an EV71 VP1 monoclonal antibody raised in mice was investigated to further identify the optimal configuration of VP1.…”

Section: Introductionmentioning

confidence: 99%

Human IgG Fc promotes expression, secretion and immunogenicity of enterovirus 71 VP1 protein

Zhang²

2016

J Biomed Res

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Enterovirus (EV71) can cause severe neurological diseases, but the underlying pathogenesis remains unclear. The capsid protein, viral protein 1 (VP1), plays a critical role in the pathogenicity of EV71. High level expression and secretion of VP1 protein are necessary for structure, function and immunogenicity in its natural conformation. In our previous studies, 5 codon-optimized VP1 DNA vaccines, including wt-VP1, tPA-VP1, VP1-d, VP1-hFc and VP1-mFc, were constructed and analyzed. They expressed VP1 protein, but the levels of secretion and immunogenicity of these VP1 constructs were significantly different (P<0.05). In this study, we further investigated the protein levels of these constructs and determined that all of these constructs expressed VP1 protein. The secretion level was increased by including a tPA leader sequence, which was further increased by fusing human IgG Fc (hFc) to VP1. VP1-hFc demonstrated the most potent immunogenicity in mice. Furthermore, hFc domain could be used to purify VP1-hFc protein for additional studies.

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Expression and immunogenicity of novel subunit enterovirus 71 VP1 antigens

Cited by 7 publications

References 23 publications

Cell Surface Nucleolin Facilitates Enterovirus 71 Binding and Infection

Cell Surface Nucleolin Facilitates Enterovirus 71 Binding and Infection

VP2 Dominated CD4+ T Cell Responses against Enterovirus 71 and Cross-Reactivity against Coxsackievirus A16 and Polioviruses in a Healthy Population

Human IgG Fc promotes expression, secretion and immunogenicity of enterovirus 71 VP1 protein

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