title: A plant-produced antigen elicits potent immune responses against West Nile virus in mice.
abstract: 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 of Nicotiana benthamiana by 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  μ g 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  μ g 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.
authors.affiliation: The Biodesign Institute, Arizona State University, 1001 S. McAllister Avenue, Tempe, AZ 85287, USA.
authors.forename: Junyun
authors.initials: J
authors.lastname: He
authors.affiliation: The Biodesign Institute, Arizona State University, 1001 S. McAllister Avenue, Tempe, AZ 85287, USA.
authors.forename: Li
authors.initials: L
authors.lastname: Peng
authors.affiliation: The Biodesign Institute, Arizona State University, 1001 S. McAllister Avenue, Tempe, AZ 85287, USA.
authors.forename: Huafang
authors.identifier: 0000-0002-3812-7627
authors.initials: H
authors.lastname: Lai
authors.affiliation: The Biodesign Institute, Arizona State University, 1001 S. McAllister Avenue, Tempe, AZ 85287, USA ; School of Life Sciences, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Jonathan
authors.identifier: 0000-0003-4145-7042
authors.initials: J
authors.lastname: Hurtado
authors.affiliation: The Biodesign Institute, Arizona State University, 1001 S. McAllister Avenue, Tempe, AZ 85287, USA ; School of Life Sciences, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Jake
authors.identifier: 0000-0002-1285-6290
authors.initials: J
authors.lastname: Stahnke
authors.affiliation: The Biodesign Institute, Arizona State University, 1001 S. McAllister Avenue, Tempe, AZ 85287, USA ; School of Life Sciences, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Chen
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000941:Antigens / Q000096:biosynthesis* / Q000276:immunology; D002736:Chloroplasts / Q000235:genetics / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000235:genetics; D006801:Humans; D051379:Mice; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; D017434:Protein Structure, Tertiary / Q000235:genetics; D014612:Vaccines / Q000096:biosynthesis* / Q000235:genetics; D014759:Viral Envelope Proteins / Q000096:biosynthesis* / Q000235:genetics; D014901:West Nile Fever / Q000276:immunology / Q000517:prevention & control*; D014902:West Nile virus / Q000235:genetics* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000941:Antigens; D014612:Vaccines; D014759:Viral Envelope Proteins