scholarly journals A Plant-Produced Antigen Elicits Potent Immune Responses against West Nile Virus in Mice

2014 ◽  
Vol 2014 ◽  
pp. 1-10 ◽  
Author(s):  
Junyun He ◽  
Li Peng ◽  
Huafang Lai ◽  
Jonathan Hurtado ◽  
Jake Stahnke ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Immune Responses ◽  
Viral Vector ◽  
Specific Binding ◽  
Low Cost ◽  
Expression System ◽  
Conformational Epitope ◽  
West Nile ◽  
Domain Iii ◽  
Subcutaneous Immunization

We described the rapid production of the domain III (DIII) of the envelope (E) protein in plants as a vaccine candidate for West Nile Virus (WNV). Using various combinations of vector modules of a deconstructed viral vector expression system, DIII was produced in three subcellular compartments in leaves ofNicotiana benthamianaby transient expression. DIII expressed at much higher levels when targeted to the endoplasmic reticulum (ER) than that targeted to the chloroplast or the cytosol, with accumulation level up to 73 μg DIII per gram of leaf fresh weight within 4 days after infiltration. Plant ER-derived DIII was soluble and readily purified to > 95% homogeneity without the time-consuming process of denaturing and refolding. Further analysis revealed that plant-produced DIII was processed properly and demonstrated specific binding to an anti-DIII monoclonal antibody that recognizes a conformational epitope. Furthermore, subcutaneous immunization of mice with 5 and 25 μg of purified DIII elicited a potent systemic response. This study provided the proof of principle for rapidly producing immunogenic vaccine candidates against WNV in plants with low cost and scalability.

scholarly journals Plant-Produced Antigen Displaying Virus-Like Particles Evokes Potent Antibody Responses against West Nile Virus in Mice

Vaccines ◽  
2021 ◽  
Vol 9 (1) ◽  
pp. 60
Author(s):  
Junyun He ◽  
Huafang Lai ◽  
Adrian Esqueda ◽  
Qiang Chen
Keyword(s):  
West Nile Virus ◽  
Fusion Protein ◽  
Nicotiana Benthamiana ◽  
Specific Binding ◽  
Low Cost ◽  
Gradient Centrifugation ◽  
Conformational Epitope ◽  
Protein Domain ◽  
Core Antigen ◽  
West Nile

In this study, we developed a hepatitis B core antigen (HBcAg)-based virus-like particle (VLP) that displays the West Nile virus (WNV) Envelope protein domain III (wDIII) as a vaccine candidate for WNV. The HBcAg-wDIII fusion protein was quickly produced in Nicotiana benthamiana plants and reached a high expression level of approximately 1.2 mg of fusion protein per gram of leaf fresh weight within six days post gene infiltration. Electron microscopy and gradient centrifugation analysis indicated that the introduction of wDIII did not interfere with VLP formation and HBcAg-wDIII successfully assembled into VLPs. HBcAg-wDIII VLPs can be easily purified in large quantities from Nicotiana benthamiana leaves to >95% homogeneity. Further analysis revealed that the wDIII was displayed properly and demonstrated specific binding to an anti-wDIII monoclonal antibody that recognizes a conformational epitope of wDIII. Notably, HBcAg-wDIII VLPs were shown to be highly immunogenic and elicited potent humoral responses in mice with antigen-specific IgG titers equivalent to that of protective wDIII antigens in previous studies. Thus, our wDIII-based VLP vaccine offers an attractive option for developing effective, safe, and low-cost vaccines against WNV.

scholarly journals A Recombinant Influenza A Virus Expressing Domain III of West Nile Virus Induces Protective Immune Responses against Influenza and West Nile Virus

PLoS ONE ◽  
2011 ◽  
Vol 6 (4) ◽  
pp. e18995 ◽  
Author(s):  
Byron E. E. Martina ◽  
Petra van den Doel ◽  
Penelope Koraka ◽  
Geert van Amerongen ◽  
Gunther Spohn ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Immune Responses ◽  
Influenza A Virus ◽  
Influenza A ◽  
West Nile ◽  
Domain Iii

scholarly journals A Novel System for Rapid and Cost-Effective Production of Detection and Diagnostic Reagents of West Nile Virus in Plants

2012 ◽  
Vol 2012 ◽  
pp. 1-10 ◽  
Author(s):  
Junyun He ◽  
Huafang Lai ◽  
Christopher Brock ◽  
Qiang Chen
Keyword(s):  
West Nile Virus ◽  
Posttranslational Modifications ◽  
Protein Production ◽  
Low Cost ◽  
Expression System ◽  
Cost Effective ◽  
West Nile ◽  
Economical System ◽  
High Scalability ◽  
Detection And Diagnosis

The threat of West Nile virus (WNV) epidemics necessitates the development of a technology platform that can produce reagents to support detection and diagnosis rapidly and inexpensively. A plant expression system is attractive for protein production due to its low-cost and high-scalability nature and its ability to make appropriate posttranslational modifications. Here, we investigated the feasibility of using plants to produce two WNV detection and diagnostic reagents to address the current cost and scalability issues. We demonstrated that WNV DIII antigen and E16 monoclonal antibody are rapidly produced at high levels in two plant species and are easily purified. Furthermore, they are effective in identifying WNV and in detecting human IgM response to WNV infection. E16 mAb does not cross-react with other flaviviruses, therefore, is valuable for improving diagnostic accuracy. This study provides a proof of principle for using plants as a robust and economical system to produce diagnostic reagents for arboviruses.

scholarly journals West Nile Virus Population Structure, Injury, and Interferon-Stimulated Gene Expression in the Brain From a Fatal Case of Encephalitis

2015 ◽  
Vol 3 (1) ◽  
Author(s):  
Nathan D. Grubaugh ◽  
Aaron Massey ◽  
Katherine D. Shives ◽  
Mark D. Stenglein ◽  
Gregory D. Ebel ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Immune Responses ◽  
Selection Pressure ◽  
Fatal Case ◽  
Brain Regions ◽  
Human Case ◽  
Innate Immune ◽  
Innate Immune Responses ◽  
West Nile ◽  
Acute Encephalitis

Abstract Background.  West Nile virus (WNV) infection in humans can result in severe, acute encephalitis typically involving subcortical gray matter brain regions. West Nile virus replication within specific human brain regions from a human case of acute encephalitis has not been studied. Methods.  We describe a fatal case of WNV encephalitis in which we obtained tissue from specific brain regions at autopsy to evaluate viral-host interactions using next-generation sequencing and immunohistochemistry analysis. Results.  We found that WNV populations in the injured subcortical brain regions exhibited increased amino acid variation and increased expression of specific interferon genes compared with cortical tissues despite similar viral burden. Conclusions.  These observational, patient-based data suggest that neuronal injury and the strength of viral selection pressure may be associated with the level of the innate immune responses. Further studies in human and animal models evaluating the role of innate immune responses on injury patterns and viral selection pressure are needed.

scholarly journals Immune responses to an attenuated West Nile virus NS4B-P38G mutant strain

Vaccine ◽  
2011 ◽  
Vol 29 (29-30) ◽  
pp. 4853-4861 ◽  
Author(s):  
Thomas Welte ◽  
Guorui Xie ◽  
Jason A. Wicker ◽  
Melissa C. Whiteman ◽  
Li Li ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Immune Responses ◽  
Mutant Strain ◽  
West Nile
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