Dengue-2 virus-like particle (VLP) based vaccine elicits the highest titers of neutralizing antibodies when produced at reduced temperature

Abstract Purpose of Review Treatment of equine insect bite hypersensitivity (IBH) needs to be improved. Allergen-specific immunotherapy (ASIT), the only curative treatment of allergy, currently has only a limited efficacy for treatment of IBH. This review highlights the latest findings in prophylactic and therapeutic strategies. Recent Findings Prophylactic vaccination against IBH using recombinant Culicoides allergen has been developed in unexposed Icelandic horses and is ready to be tested. Therapeutic virus-like particle (VLP)–based vaccines targeting equine interleukin- (IL-) 5 or IL-31 improved clinical signs of IBH by induction of anti-cytokine antibodies thus reducing eosinophil counts or allergic pruritus, respectively. Summary First studies for development of ASIT using pure r-Culicoides allergens have yielded promising results and need now to be tested in clinical studies for both prevention and treatment of IBH. Therapeutic vaccines inducing neutralizing antibodies against IL-5 or IL-31 will be valuable future treatments for reduction of clinical signs of IBH.

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Effects of the Japanese Encephalitis Virus Genotype V-Derived Sub-Viral Particles on the Immunogenicity of the Vaccine Characterized by a Novel Virus-Like Particle-Based Assay

Vaccines ◽

10.3390/vaccines7030081 ◽

2019 ◽

Vol 7 (3) ◽

pp. 81

Author(s):

Sarah Honjo ◽

Michiaki Masuda ◽

Tomohiro Ishikawa

Keyword(s):

Japanese Encephalitis Virus ◽

Japanese Encephalitis ◽

Neutralizing Antibodies ◽

Encephalitis Virus ◽

Virus Genotype ◽

Virus Like Particle ◽

Viral Particles ◽

Geographical Regions ◽

Novel Virus ◽

Genotype V

Japanese encephalitis virus (JEV) is classified into five genotypes labelled I through V. Although the genotype V (GV) JEV was originally found and had apparently been limited in Malaysia for more than 50 years, its emergence in Korea and China has recently been reported. Therefore, the GV JEV might be spreading over new geographical regions as a cause of potential public health problems. However, it is unknown whether the currently available JEV vaccines are effective against the emerging GV strains. To investigate this issue, a novel virus-like particle-based neutralizing assay was developed in this study. By using this assay, the inactivated JEV vaccine used in Japan and the recombinant sub-viral particles (SVPs) bearing the E protein of the GV Muar strain were characterized for the immunogenicity against the GV JEV. Although the inactivated vaccine alone failed to elicit a detectable level of neutralizing antibodies against the GV JEV, the vaccine added with the Muar-derived SVPs induced relatively high titers of neutralizing antibodies, associated with the efficient Th1 immune responses, against the GV JEV. The results indicate that addition of the GV JEV-derived antigens may be useful for developing the vaccine that is universally effective against JEV including the emerging GV strains.

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Virus-Like Particle Based Vaccines Elicit Neutralizing Antibodies against the HIV-1 Fusion Peptide

Vaccines ◽

10.3390/vaccines8040765 ◽

2020 ◽

Vol 8 (4) ◽

pp. 765

Author(s):

Alemu Tekewe Mogus ◽

Lihong Liu ◽

Manxue Jia ◽

Diane T. Ajayi ◽

Kai Xu ◽

...

Keyword(s):

Neutralizing Antibodies ◽

Vaccination Strategy ◽

Fusion Peptide ◽

Linear Epitope ◽

Broadly Neutralizing Antibodies ◽

Virus Like Particle ◽

Boost Immunization ◽

Potential Vaccine ◽

Hiv Envelope ◽

Hiv 1

Broadly neutralizing antibodies (bnAbs) isolated from HIV-infected individuals delineate vulnerable sites on the HIV envelope glycoprotein that are potential vaccine targets. A linear epitope within the N-terminal region of the HIV-1 fusion peptide (FP8) is the primary target of VRC34.01, a bnAb that neutralizes ~50% of primary HIV isolates. FP8 has attracted attention as a potential HIV vaccine target because it is a simple linear epitope. Here, platform technologies based on RNA bacteriophage virus-like particles (VLPs) were used to develop multivalent vaccines targeting the FP8 epitope. Both recombinant MS2 VLPs displaying the FP8 peptide and Qβ VLPs displaying chemically conjugated FP8 peptide induced high titers of FP8-specific antibodies in mice. Moreover, a heterologous prime-boost-boost regimen employing the two FP8-VLP vaccines and native envelope trimer was the most effective approach for eliciting HIV-1 neutralizing antibodies. Given the potent immunogenicity of VLP-based vaccines, this vaccination strategy—inspired by bnAb-guided epitope mapping, VLP bioengineering, and prime-boost immunization approaches—may be a useful strategy for eliciting bnAb responses against HIV.

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Improved Efficiency of a Salmonella-Based Vaccine against Human Papillomavirus Type 16 Virus-Like Particles Achieved by Using a Codon-Optimized Version of L1

Journal of Virology ◽

10.1128/jvi.78.23.12901-12909.2004 ◽

2004 ◽

Vol 78 (23) ◽

pp. 12901-12909 ◽

Cited By ~ 38

Author(s):

David Baud ◽

Françoise Ponci ◽

Martine Bobst ◽

Pierre De Grandi ◽

Denise Nardelli-Haefliger

Keyword(s):

Neutralizing Antibodies ◽

Hpv Infection ◽

Oral Immunization ◽

Human Papillomaviruses ◽

Effective Vaccine ◽

Virus Like Particle ◽

Serovar Typhimurium ◽

New Generation

ABSTRACT Cervical cancer results from cervical infection by human papillomaviruses (HPVs), especially HPV16. An effective vaccine against these HPVs is expected to have a dramatic impact on the incidence of this cancer and its precursor lesions. The leading candidate, a subunit prophylactic HPV virus-like particle (VLP) vaccine, can protect women from HPV infection. An alternative improved vaccine that avoids parenteral injection, that is efficient with a single dose, and that induces mucosal immunity might greatly facilitate vaccine implementation in different settings. In this study, we have constructed a new generation of recombinant Salmonella organisms that assemble HPV16 VLPs and induce high titers of neutralizing antibodies in mice after a single nasal or oral immunization with live bacteria. This was achieved through the expression of a HPV16 L1 capsid gene whose codon usage was optimized to fit with the most frequently used codons in Salmonella. Interestingly, the high immunogenicity of the new recombinant bacteria did not correlate with an increased expression of L1 VLPs but with a greater stability of the L1-expressing plasmid in vitro and in vivo in absence of antibiotic selection. Anti-HPV16 humoral and neutralizing responses were also observed with different Salmonella enterica serovar Typhimurium strains whose attenuating deletions have already been shown to be safe after oral vaccination of humans. Thus, our findings are a promising improvement toward a vaccine strain that could be tested in human volunteers.

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Structure, Immunogenicity, and Protective Mechanism of an Engineered Enterovirus 71-Like Particle Vaccine Mimicking 80S Empty Capsid

Journal of Virology ◽

10.1128/jvi.01330-17 ◽

2017 ◽

Vol 92 (1) ◽

Cited By ~ 3

Author(s):

Xiaoli Wang ◽

Zhiqiang Ku ◽

Xiang Zhang ◽

Xiaohua Ye ◽

Jinhuan Chen ◽

...

Keyword(s):

Neutralizing Antibodies ◽

Enterovirus 71 ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Asia Pacific ◽

Protective Mechanism ◽

Particle Assembly ◽

Virus Like Particle ◽

Empty Capsid ◽

Foot And Mouth

ABSTRACTEnterovirus 71 (EV71) is the major causative agent of severe hand, foot, and mouth disease, which affects millions of young children in the Asia-Pacific region annually. In this study, we engineered a novel EV71 virus-like particle (VLP) that lacks VP4 (therefore designated VLPΔVP4) and investigated its structure, antigenicity, and vaccine potential. The cryo-electron microscopy (cryo-EM) structure of VLPΔVP4was reconstructed to 3.71-Å resolution. Results from structural and biochemical analyses revealed that VLPΔVP4resembles the end product of the viral uncoating process, the 80S empty capsid. VLPΔVP4is able to elicit high-titer neutralizing antibodies and to fully protect mice against lethal viral challenge. Mechanistic studies showed that, at the cellular level, the anti-VLPΔVP4sera exert neutralization effects at both pre- and postattachment stages by inhibiting both virus attachment and internalization, and at the molecular level, the antisera can block multiple interactions between EV71 and its key receptors. Our study gives a better understanding of EV71 capsid assembly and provides important information for the design and development of new-generation vaccines for EV71, and perhaps for other enteroviruses, as well.IMPORTANCEEnterovirus 71 (EV71) infection may lead to severe hand, foot, and mouth disease, with significant morbidity and mortality. Knowledge regarding EV71 particle assembly remains limited. Here, we report the generation and characterization of a novel EV71 virus-like particle that lacks the VP4 capsid subunit protein. This particle, termed VLPΔVP4, structurally mimics the 80S empty capsid, which is the end stage of EV71 uncoating. We further show that VLPΔVP4exhibits desirable immunogenicity and protective efficacy in proof-of-concept studies. In addition, the inhibitory mechanisms of the VLPΔVP4-induced antibodies are unraveled at both the cellular and molecular levels. Our work provides the first evidence of picornaviral particle assembly in the complete absence of VP4 and identifies VLPΔVP4as an improved EV71 vaccine candidate with desirable traits. These findings not only enhance our understanding of particle assembly and uncoating of picornaviruses, but also provide important information for structure-guided vaccine design for EV71 and other enteroviruses.

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Safe and effective two-in-one replicon-and-VLP minispike vaccine for COVID-19: Protection of mice after a single immunization

PLoS Pathogens ◽

10.1371/journal.ppat.1009064 ◽

2021 ◽

Vol 17 (4) ◽

pp. e1009064

Author(s):

Alexandru A. Hennrich ◽

Bevan Sawatsky ◽

Rosalía Santos-Mandujano ◽

Dominic H. Banda ◽

Martina Oberhuber ◽

...

Keyword(s):

Neutralizing Antibodies ◽

Antigen Expression ◽

Vaccine Platform ◽

Dna And Rna ◽

Glycoprotein G ◽

Virus Like Particle ◽

Main Target ◽

Boost Immunization ◽

Vesicular Stomatitis ◽

Anchor Sequence

Vaccines of outstanding efficiency, safety, and public acceptance are needed to halt the current SARS-CoV-2 pandemic. Concerns include potential side effects caused by the antigen itself and safety of viral DNA and RNA delivery vectors. The large SARS-CoV-2 spike (S) protein is the main target of current COVID-19 vaccine candidates but can induce non-neutralizing antibodies, which might cause vaccination-induced complications or enhancement of COVID-19 disease. Besides, encoding of a functional S in replication-competent virus vector vaccines may result in the emergence of viruses with altered or expanded tropism. Here, we have developed a safe single round rhabdovirus replicon vaccine platform for enhanced presentation of the S receptor-binding domain (RBD). Structure-guided design was employed to build a chimeric minispike comprising the globular RBD linked to a transmembrane stem-anchor sequence derived from rabies virus (RABV) glycoprotein (G). Vesicular stomatitis virus (VSV) and RABV replicons encoding the minispike not only allowed expression of the antigen at the cell surface but also incorporation into the envelope of secreted non-infectious particles, thus combining classic vector-driven antigen expression and particulate virus-like particle (VLP) presentation. A single dose of a prototype replicon vaccine complemented with VSV G, VSVΔG-minispike-eGFP (G), stimulated high titers of SARS-CoV-2 neutralizing antibodies in mice, equivalent to those found in COVID-19 patients, and protected transgenic K18-hACE2 mice from COVID-19-like disease. Homologous boost immunization further enhanced virus neutralizing activity. The results demonstrate that non-spreading rhabdovirus RNA replicons expressing minispike proteins represent effective and safe alternatives to vaccination approaches using replication-competent viruses and/or the entire S antigen.

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Virus-Like Particle Vaccine Containing the F Protein of Respiratory Syncytial Virus Confers Protection without Pulmonary Disease by Modulating Specific Subsets of Dendritic Cells and Effector T Cells

Journal of Virology ◽

10.1128/jvi.02018-15 ◽

2015 ◽

Vol 89 (22) ◽

pp. 11692-11705 ◽

Cited By ~ 27

Author(s):

Ki-Hye Kim ◽

Young-Tae Lee ◽

Hye Suk Hwang ◽

Young-Man Kwon ◽

Min-Chul Kim ◽

...

Keyword(s):

Weight Loss ◽

T Cells ◽

Dendritic Cells ◽

Respiratory Syncytial Virus ◽

Neutralizing Antibodies ◽

Immune Correlates ◽

Virus Like Particle ◽

Tnf Α ◽

Ifn Γ ◽

Syncytial Virus

ABSTRACTThere is no licensed vaccine against respiratory syncytial virus (RSV) since the failure of formalin-inactivated RSV (FI-RSV) due to its vaccine-enhanced disease. We investigated immune correlates conferring protection without causing disease after intranasal immunization with virus-like particle vaccine containing the RSV fusion protein (F VLP) in comparison to FI-RSV and live RSV. Upon RSV challenge, FI-RSV immune mice showed severe weight loss, eosinophilia, and histopathology, and RSV reinfection also caused substantial RSV disease despite their viral clearance. In contrast, F VLP immune mice showed least weight loss and no sign of histopathology and eosinophilia. High levels of interleukin-4-positive (IL-4+) and tumor necrosis factor alpha-positive (TNF-α+) CD4+T cells were found in FI-RSV immune mice, whereas gamma interferon-positive (IFN-γ+) and TNF-α+CD4+T cells were predominantly detected in live RSV-infected mice. More importantly, in contrast to FI-RSV and live RSV that induced higher levels of CD11b+dendritic cells, F VLP immunization induced CD8α+and CD103+dendritic cells, as well as F-specific IFN-γ+and TNF-α+CD8+T cells. These results suggest that F VLP can induce protection without causing pulmonary RSV disease by inducing RSV neutralizing antibodies, as well as modulating specific subsets of dendritic cells and CD8 T cell immunity.IMPORTANCEIt has been a difficult challenge to develop an effective and safe vaccine against respiratory syncytial virus (RSV), a leading cause of respiratory disease. Immune correlates conferring protection but preventing vaccine-enhanced disease remain poorly understood. RSV F virus-like particle (VLP) would be an efficient vaccine platform conferring protection. Here, we investigated the protective immune correlates without causing disease after intranasal immunization with RSV F VLP in comparison to FI-RSV and live RSV. In addition to inducing RSV neutralizing antibodies responsible for clearing lung viral loads, we show that modulation of specific subsets of dendritic cells and CD8 T cells producing T helper type 1 cytokines are important immune correlates conferring protection but not causing vaccine-enhanced disease.

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

Journal of Virology ◽

10.1128/jvi.01848-13 ◽

2013 ◽

Vol 88 (1) ◽

pp. 72-81 ◽

Cited By ~ 51

Author(s):

X. Ye ◽

Z. Ku ◽

Q. Liu ◽

X. Wang ◽

J. Shi ◽

...

Keyword(s):

Neutralizing Antibodies ◽

Enterovirus 71 ◽

Chimeric Virus ◽

Virus Like Particle

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Zika virus-like particle vaccine protects AG129 mice and rhesus macaques against Zika virus

PLoS Neglected Tropical Diseases ◽

10.1371/journal.pntd.0009195 ◽

2021 ◽

Vol 15 (3) ◽

pp. e0009195

Author(s):

Lo Vang ◽

Christopher S. Morello ◽

Jason Mendy ◽

Danielle Thompson ◽

Darly Manayani ◽

...

Keyword(s):

Zika Virus ◽

Mammalian Cells ◽

Neutralizing Antibodies ◽

Rhesus Macaques ◽

High Titer ◽

Neutralization Assay ◽

Dose Titration ◽

Development Programs ◽

Structural Protein ◽

Virus Like Particle

Background Zika virus (ZIKV), a mosquito-borne flavivirus, is a re-emerging virus that constitutes a public health threat due to its recent global spread, recurrent outbreaks, and infections that are associated with neurological abnormalities in developing fetuses and Guillain-Barré syndrome in adults. To date, there are no approved vaccines against ZIKV infection. Various preclinical and clinical development programs are currently ongoing in an effort to bring forward a vaccine for ZIKV. Methodology/Principle findings We have developed a ZIKV vaccine candidate based on Virus-Like-Particles (VLPs) produced in HEK293 mammalian cells using the prM (a precursor to M protein) and envelope (E) structural protein genes from ZIKV. Transient transfection of cells via plasmid and electroporation produced VLPs which were subsequently purified by column chromatography yielding approximately 2mg/L. Initially, immunogenicity and efficacy were evaluated in AG129 mice using a dose titration of VLP with and without Alhydrogel 2% (alum) adjuvant. We found that VLP with and without alum elicited ZIKV-specific serum neutralizing antibodies (nAbs) and that titers correlated with protection. A follow-up immunogenicity and efficacy study in rhesus macaques was performed using VLP formulated with alum. Multiple neutralization assay methods were performed on immune sera including a plaque reduction neutralization test, a microneutralization assay, and a Zika virus Renilla luciferase neutralization assay. All of these assays indicate that following immunization, VLP induces high titer nAbs which correlate with protection against ZIKV challenge. Conclusions/Significance These studies confirm that ZIKV VLPs could be efficiently generated and purified. Upon VLP immunization, in both mice and NHPs, nAb was induced that correlate with protection against ZIKV challenge. These studies support translational efforts in developing a ZIKV VLP vaccine for evaluation in human clinical trials.

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Newcastle Disease Virus-Like Particles Containing Respiratory Syncytial Virus G Protein Induced Protection in BALB/c Mice, with No Evidence of Immunopathology

Journal of Virology ◽

10.1128/jvi.01709-09 ◽

2009 ◽

Vol 84 (2) ◽

pp. 1110-1123 ◽

Cited By ~ 90

Author(s):

Matthew R. Murawski ◽

Lori W. McGinnes ◽

Robert W. Finberg ◽

Evelyn A. Kurt-Jones ◽

Michael J. Massare ◽

...

Keyword(s):

Respiratory Syncytial Virus ◽

Newcastle Disease Virus ◽

G Protein ◽

Disease Virus ◽

Newcastle Disease ◽

Neutralizing Antibodies ◽

Respiratory Infections ◽

Chimeric Protein ◽

Virus Like Particle ◽

Syncytial Virus

ABSTRACT Respiratory syncytial virus (RSV) is the leading cause of serious respiratory infections in children as well as a serious cause of disease in elderly and immunosuppressed populations. There are no licensed vaccines available to prevent RSV disease. We have developed a virus-like particle (VLP) vaccine candidate for protection from RSV. The VLP is composed of the NP and M proteins of Newcastle disease virus (NDV) and a chimeric protein containing the cytoplasmic and transmembrane domains of the NDV HN protein and the ectodomain of the human RSV G protein (H/G). Immunization of mice with 10 or 40 μg total VLP-H/G protein by intraperitoneal or intramuscular inoculation stimulated antibody responses to G protein which were as good as or better than those stimulated by comparable amounts of UV-inactivated RSV. Immunization of mice with two doses or even a single dose of these particles resulted in the complete protection of mice from RSV replication in the lungs. Immunization with these particles induced neutralizing antibodies with modest titers. Upon RSV challenge of VLP-H/G-immunized mice, no enhanced pathology in the lungs was observed, although lungs of mice immunized in parallel with formalin-inactivated RSV (FI-RSV) showed the significant pathology that has previously been documented after immunization with FI-RSV. Thus, the VLP-H/G candidate vaccine was immunogenic in BALB/c mice and prevented replication of RSV in murine lungs, with no evidence of immunopathology. These data support further development of virus-like particle vaccine candidates for protection against RSV.

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