Expression and purification of norovirus virus like particles in Escherichia coli and their immunogenicity in mice

Huo, Yuqi; Wan, Xin; Ling, Tong; Wu, Jie; Wang, Wenhui; Shen, Shuo

doi:10.1016/j.molimm.2017.07.014

Cited by 27 publications

(20 citation statements)

References 22 publications

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“…VLPs are multi-protein structures that resemble the organization and morphology of the native virus, but contain no genetic material and are therefore noninfectious. VLPs are not only typically produced by recombinant baculovirus, but also can be produced by E. coli, Pichia pastoris (yeast), and plants, which, when optimized, may lower the cost of VLP production [56–59]. P particles are developed to resemble the P domain of norovirus, which is the part of the virus that binds to HBGAs [60,61].…”

Section: Vaccine Developmentmentioning

confidence: 99%

Progress on norovirus vaccine research: public health considerations and future directions

Mattison

Cardemil

Hall

2018

Expert Review of Vaccines

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Noroviruses are the leading cause of foodborne illness worldwide, account for approximately one-fifth of acute gastroenteritis (AGE) cases globally, and cause a substantial economic burden. Candidate norovirus vaccines are in development, but there is currently no licensed vaccine. Areas covered: Noroviruses cause approximately 684 million cases and 212,000 deaths per year across all age groups, though burden estimates vary by study and region. Challenges to vaccine research include substantial and rapidly evolving genetic diversity, short-term and homotypic immunity to infection, and the absence of a single, well-established correlate of protection. Nonetheless, several norovirus vaccine candidates are currently in development, utilizing virus-like particles (VLPs), P particles, and recombinant adenoviruses. Of these, a bivalent GI.1/GII.4 VLP-based intramuscular vaccine (Phase IIb) and GI.1 oral vaccine (Phase I) are in clinical trials. Expert Commentary: A norovirus vaccine should target high-risk populations, including the young and the elderly, and protect them against the most common circulating norovirus strains. A norovirus vaccine would be a powerful tool in the prevention and control of norovirus while lessening the burden of AGE worldwide. However, more robust burden and cost estimates are needed to justify investments in and guide norovirus vaccine development.

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Section: Vaccine Developmentmentioning

confidence: 99%

Progress on norovirus vaccine research: public health considerations and future directions

Mattison

Cardemil

Hall

2018

Expert Review of Vaccines

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“…Research has indicated that structural proteins of the virus not only play an important role in viral invasion and replication but also have excellent immunogenicity and that they are frequently selected as candidates for the development of therapeutic antibodies (Huo et al, 2018;Li et al, 2014;Stachyra et al, 2016;Wang et al, 2017). Compared with vertebrate immune systems, honey bee immune pathways lack acquired immunity (Bull et al, 2012) and cannot directly generate specific antibodies in response to an antigen.…”

Section: Discussionmentioning

confidence: 99%

Codon optimization, expression in Escherichia coli, and immunogenicity analysis of deformed wing virus (DWV) structural protein

Fei

Guo

Fan

et al. 2020

PeerJ

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Background Deformed wing virus (DWV) is a serious threat to honey bees (Apis mellifera) and is considered a major cause of elevated losses of honey bee colonies. However, lack of information on the immunogenicity of DWV structural proteins has hindered the development of effective biocontrol drugs. Methods We optimized the VP1, VP2 and VP3 codons of DWV surface capsid protein genes on the basis of an Escherichia coli codon bias, and the optimized genes of roVP1, roVP2 and roVP3 were separately expressed in E. coli and purified. Next, the three recombinant proteins of roVP1, roVP2 and roVP3 were intramuscularly injected into BALB/c and the immunogenicity was evaluated by the levels of specific IgG and cytokines. Furthermore, anti-roVP-antisera (roVP1 or roVP2 or roVP3) from the immunized mice was incubated with DWV for injecting healthy white-eyed pupae for the viral challenge test, respectively. Results The optimized genes roVP1, roVP2 and roVP3 achieved the expression in E. coli using SDS-PAGE and Western blotting. Post-immunization, roVP2 and roVP3 exhibited higher immunogenicity than roVP1 and stimulated a stronger humoral immune response in the mice, which showed that the recombinant proteins of roVP3 and roVP2 induced a specific immune response in the mice. In the challenge test, data regarding quantitative real-time RT-PCR (qRT-PCR) from challenged pupae showed that the level of virus copies in the recombinant protein groups was significantly lower than that of the virus-only group at 96 h post-inoculation (P < 0.05). Among them, the degree of neutralization using antibodies raised to the recombinant proteins are between approximately 2-fold and 4-fold and the virus copies of the roVP3 group are the lowest in the three recombinant protein groups, which indicated that specific antibodies against recombinant proteins roVP1, roVP2 and roVP3 of DWV could neutralize DWV to reduce the virus titer in the pupae. Collectively, these results demonstrated that the surface capsid protein of DWV acted as candidates for the development of therapeutic antibodies against the virus.

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Section: Discussionmentioning

confidence: 99%

Peer Review #1 of "Codon optimization, expression in Escherichia coli, and immunogenicity analysis of deformed wing virus (DWV) structural protein (v0.1)"

2020

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Background: Deformed wing virus (DWV) is a serious threat to honey bees (Apis mellifera) and is considered a major cause of elevated losses of honey bee colonies. However, lack of information on the immunogenicity of DWV structural proteins has hindered the development of effective biocontrol drugs. Methods: We optimized the VP1, VP2, and VP3 codons of DWV surface capsid protein genes on the basis of an Escherichia coli codon bias, and the optimized genes of roVP1, roVP2, and roVP3 were separately expressed in E. coli and purified. Next, the three recombinant proteins of roVP1, roVP2, and roVP3 were intramuscularly injected into BALB/c, and the immunogenicity was evaluated by the levels of specific IgG and cytokines. Furthermore, anti-roVP-antisera (roVP1 or roVP2 or roVP3) from the immunized mice was incubated with DWV for injecting healthy white-eyed pupae for the viral challenge test, respectively. Results: The optimized genes roVP1, roVP2, and roVP3 achieved the expression in E. coli using SDS-PAGE and Western blotting. Post-immunization, roVP2 and roVP3 exhibited higher immunogenicity than roVP1 and stimulated a stronger humoral immune response in the mice, which showed that the recombinant proteins of roVP3 and roVP2 induced a specific immune response in the mice. In the challenge test, data regarding quantitative real-time RT-PCR (qRT-PCR) from challenged pupae showed that the level of virus copies in the recombinant protein groups was significantly lower than that of the virus-only group at 96 h post-inoculation (P < 0.05). Among them, the degree of neutralisation using antibodies raised to the recombinant proteins are between approximately 2-fold and 4-fold, and the virus copies of the roVP3 group are the lowest in the three recombinant protein groups, which indicated that specific antibodies against recombinant proteins roVP1, roVP2, and roVP3 of DWV could neutralize DWV to reduce the virus titer in the pupae. Collectively, these results demonstrated that the surface capsid protein of DWV acted as candidates for

show abstract

Expression and purification of norovirus virus like particles in Escherichia coli and their immunogenicity in mice

Cited by 27 publications

References 22 publications

Progress on norovirus vaccine research: public health considerations and future directions

Progress on norovirus vaccine research: public health considerations and future directions

Codon optimization, expression in Escherichia coli, and immunogenicity analysis of deformed wing virus (DWV) structural protein

Peer Review #1 of "Codon optimization, expression in Escherichia coli, and immunogenicity analysis of deformed wing virus (DWV) structural protein (v0.1)"

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