Immunogenicity and efficacy of two types of West Nile virus-like particles different in size and maturation as a second-generation vaccine candidate

ABSTRACT A trans-packaging system for West Nile virus (WNV) subgenomic replicon RNAs (repRNAs), deleted for the structural coding region, was developed. WNV repRNAs were efficiently encapsidated by the WNV C/prM/E structural proteins expressed in trans from replication-competent, noncytopathic Sindbis virus-derived RNAs. Infectious virus-like particles (VLPs) were produced in titers of up to 109 infectious units/ml. WNV VLPs established a single round of infection in a variety of different cell lines without production of progeny virions. The infectious properties of WNV and VLPs were indistinguishable when efficiencies of infection of a number of different cell lines and inhibition of infection by neutralizing antibodies were determined. To investigate the usefulness of VLPs to address biological questions in vivo, Culex pipiens quinquefasciatus mosquitoes were orally and parenterally infected with VLPs, and dissected tissues were analyzed for WNV antigen expression. Antigen-positive cells in midguts of orally infected mosquitoes were detected as early as 2 days postinfection and as late as 8 days. Intrathoracic inoculation of VLPs into mosquitoes demonstrated a dose-dependent pattern of infection of secondary tissues and identified fat body, salivary glands, tracheal cells, and midgut muscle as susceptible WNV VLP infection targets. These results demonstrate that VLPs can serve as a valuable tool for the investigation of tissue tropism during the early stages of infection, where virus spread and the need for biosafety level 3 containment complicate the use of wild-type virus.

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Purification and concentration of non-infectious West Nile virus-like particles and infectious virions using a pseudo-affinity Cellufine Sulfate column

Journal of Virological Methods ◽

10.1016/j.jviromet.2011.03.021 ◽

2011 ◽

Vol 174 (1-2) ◽

pp. 131-135 ◽

Cited By ~ 20

Author(s):

Naohiro Ohtaki ◽

Hidehiro Takahashi ◽

Keiko Kaneko ◽

Yasuyuki Gomi ◽

Toyokazu Ishikawa ◽

...

Keyword(s):

West Nile Virus ◽

West Nile ◽

Virus Like Particles

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Transcellular transport of West Nile virus-like particles across human endothelial cells depends on residues 156 and 159 of envelope protein

BMC Microbiology ◽

10.1186/1471-2180-10-165 ◽

2010 ◽

Vol 10 (1) ◽

pp. 165 ◽

Cited By ~ 25

Author(s):

Rie Hasebe ◽

Tadaki Suzuki ◽

Yoshinori Makino ◽

Manabu Igarashi ◽

Satoko Yamanouchi ◽

...

Keyword(s):

Endothelial Cells ◽

West Nile Virus ◽

Envelope Protein ◽

Human Endothelial Cells ◽

West Nile ◽

Transcellular Transport ◽

Virus Like Particles

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Construction and characterization of a second-generation pseudoinfectious West Nile virus vaccine propagated using a new cultivation system

Vaccine ◽

10.1016/j.vaccine.2008.03.009 ◽

2008 ◽

Vol 26 (22) ◽

pp. 2762-2771 ◽

Cited By ~ 51

Author(s):

Douglas G. Widman ◽

Tomohiro Ishikawa ◽

Rafik Fayzulin ◽

Nigel Bourne ◽

Peter W. Mason

Keyword(s):

West Nile Virus ◽

Virus Vaccine ◽

Second Generation ◽

West Nile ◽

Cultivation System

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Identification of Mutated Cyclization Sequences That Permit Efficient Replication of West Nile Virus Genomes: Use in Safer Propagation of a Novel Vaccine Candidate

Journal of Virology ◽

10.1128/jvi.00662-08 ◽

2008 ◽

Vol 82 (14) ◽

pp. 6942-6951 ◽

Cited By ~ 24

Author(s):

Ryosuke Suzuki ◽

Rafik Fayzulin ◽

Ilya Frolov ◽

Peter W. Mason

Keyword(s):

West Nile Virus ◽

Vaccine Candidate ◽

Systematic Evaluation ◽

Genome Replication ◽

West Nile ◽

Recombinant Viruses ◽

System A ◽

Mammalian Cell Lines ◽

Efficient Replication ◽

Virus Genomes

ABSTRACT Existing live-attenuated flavivirus vaccines (LAV) could be improved by reducing their potential to recombine with naturally circulating viruses in the field. Since the highly conserved cyclization sequences (CS) found in the termini of flavivirus genomes must be complementary to each other to support genome replication, we set out to identify paired mutant CS that could support the efficient replication of LAV but would be unable to support replication in recombinant viruses harboring one wild-type (WT) CS. By systematic evaluation of paired mutated CS encoded in West Nile virus (WNV) replicons, we identified variants having single and double mutations in the 5′- and 3′-CS components that could support genome replication at WT levels. Replicons containing only the double-mutated CS in the 5′ or the 3′ ends of the genome were incapable of replication, indicating that mutated CS could be useful for constructing safer LAV. Despite the identity of the central portion of the CS in all mosquito-borne flaviviruses, viruses carrying complementary the double mutations in both the 5′- and the 3′-CS were indistinguishable from WT WNV in their replication in insect and mammalian cell lines. In addition to the utility of our novel CS pair in constructing safer LAV, we demonstrated that introduction of these mutated CS into one component of a recently described two-component genome system (A. V. Shustov, P. W. Mason, and I. Frolov, J. Virol. 81:11737-11748, 2007) enabled us to engineer a safer single-cycle WNV vaccine candidate with reduced potential for recombination during its propagation.

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Development of chimaeric West Nile virus attenuated vaccine candidate based on the Japanese encephalitis vaccine strain SA14-14-2

Journal of General Virology ◽

10.1099/vir.0.059436-0 ◽

2013 ◽

Vol 94 (12) ◽

pp. 2700-2709 ◽

Cited By ~ 12

Author(s):

Xiao-Feng Li ◽

Wei Zhao ◽

Fang Lin ◽

Qing Ye ◽

Hong-Jiang Wang ◽

...

Keyword(s):

West Nile Virus ◽

Japanese Encephalitis ◽

Vaccine Development ◽

Vaccine Candidate ◽

Genetic System ◽

Reverse Genetic System ◽

West Nile ◽

Protective Antigens ◽

Japanese Encephalitis Vaccine

Mosquito-borne flaviviruses include a large group of important human medical pathogens. Several chimaeric flaviviruses have been constructed, and show potential for vaccine development. Although Japanese encephalitis virus (JEV) live vaccine SA14-14-2 has been widely used with ideal safety and efficacy profiles, no chimaeric flavivirus based on the JEV vaccine has been described to date. Based on the reverse genetic system of the JEV vaccine SA14-14-2, a novel live chimaeric flavivirus carrying the protective antigens of West Nile virus (WNV) was constructed and recovered in this study. The resulting chimaera (ChinWNV) replicated efficiently in both mammalian and mosquito cells and possessed genetic stability after in vitro serial passaging. ChinWNV exhibited a small-plaque phenotype, and its replication was significantly restricted in mouse peripheral blood and brain compared with parental WNV. Importantly, ChinWNV was highly attenuated with regard to both neurovirulence and neuroinvasiveness in mice. Furthermore, a single ChinWNV immunization stimulated robust WNV-specific adaptive immune responses in mice, conferring significant protection against lethal WNV infection. Our results demonstrate that chimaeric flaviviruses based on the JEV vaccine can serve as a powerful platform for vaccine development, and that ChinWNV represents a potential WNV vaccine candidate that merits further development.

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TLR3- and MyD88-Dependent Signaling Differentially Influences the Development of West Nile Virus-Specific B Cell Responses in Mice following Immunization with RepliVAX WN, a Single-Cycle Flavivirus Vaccine Candidate

Journal of Virology ◽

10.1128/jvi.01469-13 ◽

2013 ◽

Vol 87 (22) ◽

pp. 12090-12101 ◽

Cited By ~ 17

Author(s):

J. Xia ◽

E. R. Winkelmann ◽

S. R. Gorder ◽

P. W. Mason ◽

G. N. Milligan

Keyword(s):

West Nile Virus ◽

B Cell ◽

Vaccine Candidate ◽

Single Cycle ◽

West Nile ◽

Cell Responses ◽

B Cell Responses ◽

Flavivirus Vaccine

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Immunoinformatics and molecular dynamics approaches: Next generation vaccine design against West Nile virus

PLoS ONE ◽

10.1371/journal.pone.0253393 ◽

2021 ◽

Vol 16 (6) ◽

pp. e0253393

Author(s):

Md Tahsin Khan ◽

Rahatul Islam ◽

Tarhima Jahan Jerin ◽

Araf Mahmud ◽

Sahara Khatun ◽

...

Keyword(s):

Molecular Dynamics ◽

West Nile Virus ◽

Dynamics Simulation ◽

West Nile ◽

Strong Immune Response ◽

Generation Vaccine ◽

Life Threatening ◽

Microbial System

West Nile Virus (WNV) is a life threatening flavivirus that causes significant morbidity and mortality worldwide. No preventive therapeutics including vaccines against WNV are available for human use. In this study, immunoinformatics approach was performed to design a multi epitope-based subunit vaccine against this deadly pathogen. Human (HLA) and Mice (H-2) allele specific potential T-cell and B-cell epitopes were shortlisted through a stringent procedure. Molecular docking showed selected epitopes that have stronger binding affinity with human TLR-4. Molecular dynamics simulation confirmed the stable nature of the docked complex. Furthermore, in silico cloning analysis ensures efficient expression of desired gene in the microbial system. Interestingly, previous studies showed that two of our selected epitopes have strong immune response against WNV. Therefore, selected epitopes could be strong vaccine candidates to prevent WNV infections in human. However, further in vitro and in vivo investigations could be strengthening the validation of the vaccine candidate against WNV.

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