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Development of Virus-Like-Particle Vaccine and Reporter Assay for Zika Virus.

作者信息

Garg Himanshu, Sedano Melina, Plata Gabrielle, Punke Erin B, Joshi Anjali

机构信息

Center of Emphasis in Infectious Diseases, Department of Biomedical Sciences, Texas Tech University Health Sciences Center, El Paso, Texas, USA

Center of Emphasis in Infectious Diseases, Department of Biomedical Sciences, Texas Tech University Health Sciences Center, El Paso, Texas, USA.

出版信息

J Virol. 2017 Sep 27;91(20). doi: 10.1128/JVI.00834-17. Print 2017 Oct 15.


DOI:10.1128/JVI.00834-17
PMID:28794019
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5625514/
Abstract

Recent worldwide outbreaks of Zika virus (ZIKV) infection and the lack of an approved vaccine raise serious concerns regarding preparedness to combat this emerging virus. We used a virus-like particle (VLP)-based approach to develop a vaccine and a microneutralization assay for ZIKV. A synthetic capsid-premembrane-envelope (C-prM-E) gene construct of ZIKV was used to generate reporter virus particles (RVPs) that package a green fluorescent protein (GFP) reporter-expressing West Nile virus (WNV) replicon. The assay was adapted to a 96-well format, similar to the plaque reduction neutralization test (PRNT), and showed high reproducibility with specific detection of ZIKV neutralizing antibodies. Furthermore, C-prM-E and prM-E VLPs were tested as vaccine candidates in mice and compared to DNA vaccination. While the ZIKV prM-E construct alone was sufficient for generating VLPs, efficient VLP production from the C-prM-E construct could be achieved in the presence of the WNV NS2B-3 protease, which cleaves C from prM, allowing virus release. Immunization studies in mice showed that VLPs generated higher neutralizing antibody titers than those with the DNA vaccines, with C-prM-E VLPs giving slightly higher titers than those with prM-E VLPs. The superiority of C-prM-E VLPs suggests that inclusion of capsid may have benefits for ZIKV and other flaviviral VLP vaccines. To facilitate the VLP platform, we generated a stable cell line expressing high levels of ZIKV prM-E proteins that constitutively produce VLPs as well as a cell line expressing ZIKV C-prM-E proteins for RVP production. While several vaccine platforms have been proposed for ZIKV, this study describes a safe, effective, and economical VLP-based vaccine against ZIKV. To address the growing Zika virus epidemic, we undertook this study with two objectives: first, to develop a safe, effective, and economical vaccine for ZIKV, and second, to develop a rapid and versatile assay to detect the anti-ZIKV immune response. We generated a cell line stably expressing ZIKV prM-E that produces large amounts of VLPs in the supernatant and a ZIKV C-prM-E cell line that produces reporter virus particles upon transfection with a GFP replicon plasmid. The prM-E VLPs induced a strong neutralizing antibody response in mice that was better when the capsid was included. VLP-based vaccines showed significantly better neutralizing antibody responses than those with their DNA counterparts. The RVP-based microneutralization assay worked similarly to the PRNT assay, with a rapid GFP readout in a 96-well format. Our VLP-based platform provides a source for a ZIKV vaccine and diagnosis that can rapidly be adapted to current outbreaks.

摘要

相似文献

[1]
Development of Virus-Like-Particle Vaccine and Reporter Assay for Zika Virus.

J Virol. 2017-9-27

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[6]
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[7]
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[8]
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Sci China Life Sci. 2025-6-3

[2]
A review on Zika vaccine development.

Pathog Dis. 2024-2-7

[3]
Identification of a critical role for ZIKV capsid α3 in virus assembly and its genetic interaction with M protein.

PLoS Negl Trop Dis. 2024-1

[4]
Zika virus prM protein contains cholesterol binding motifs required for virus entry and assembly.

Nat Commun. 2023-11-13

[5]
Generation of Human 293-F Suspension NGFR Knockout Cells Using CRISPR/Cas9 Coupled to Fluorescent Protein Expression.

Methods Mol Biol. 2023

[6]
Development of a novel virus-like particle-based vaccine for preventing tick-borne encephalitis virus infection.

Virol Sin. 2023-10

[7]
An optimized messenger RNA vaccine candidate protects non-human primates from Zika virus infection.

NPJ Vaccines. 2023-4-20

[8]
Bioluminescent and Fluorescent Reporter-Expressing Recombinant SARS-CoV-2.

Methods Mol Biol. 2022

[9]
Identification and characterization of key residues in Zika virus envelope protein for virus assembly and entry.

Emerg Microbes Infect. 2022-12

[10]
Measles-based Zika vaccine induces long-term immunity and requires NS1 antibodies to protect the female reproductive tract.

NPJ Vaccines. 2022-4-19

本文引用的文献

[1]
Zika virus-like particle (VLP) based vaccine.

PLoS Negl Trop Dis. 2017-5-8

[2]
Modified mRNA Vaccines Protect against Zika Virus Infection.

Cell. 2017-3-23

[3]
An RNA nanoparticle vaccine against Zika virus elicits antibody and CD8+ T cell responses in a mouse model.

Sci Rep. 2017-3-21

[4]
Structural Biology of the Zika Virus.

Trends Biochem Sci. 2017-3-16

[5]
A human antibody against Zika virus crosslinks the E protein to prevent infection.

Nat Commun. 2017-3-16

[6]
Exploiting virus-like particles as innovative vaccines against emerging viral infections.

J Microbiol. 2017-3

[7]
Zika virus protection by a single low-dose nucleoside-modified mRNA vaccination.

Nature. 2017-3-9

[8]
Molecular determinants of human neutralizing antibodies isolated from a patient infected with Zika virus.

Sci Transl Med. 2016-12-14

[9]
Neutralizing human antibodies prevent Zika virus replication and fetal disease in mice.

Nature. 2016-12-15

[10]
The flavivirus capsid protein: Structure, function and perspectives towards drug design.

Virus Res. 2017-1-2

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