title: Pichia pastoris-expressed dengue 3 envelope-based virus-like particles elicit predominantly domain III-focused high titer neutralizing antibodies.
abstract: Dengue poses a serious public health risk to nearly half the global population. It causes ~400 million infections annually and is considered to be one of the fastest spreading vector-borne diseases. Four distinct serotypes of dengue viruses (DENV-1, -2, -3, and -4) cause dengue disease, which may be either mild or extremely severe. Antibody-dependent enhancement (ADE), by pre-existing cross-reactive antibodies, is considered to be the major mechanism underlying severe disease. This mandates that a preventive vaccine must confer simultaneous and durable immunity to each of the four prevalent DENV serotypes. Recently, we used Pichia pastoris, to express recombinant DENV-2 E ectodomain, and found that it assembled into virus-like particles (VLPs), in the absence of prM, implicated in the elicitation of ADE-mediating antibodies. These VLPs elicited predominantly type-specific neutralizing antibodies that conferred significant protection against lethal DENV-2 challenge, in a mouse model. The current work is an extension of this approach to develop prM-lacking DENV-3 E VLPs. Our data reveal that P. pastoris-produced DENV-3 E VLPs not only preserve the antigenic integrity of the major neutralizing epitopes, but also elicit potent DENV-3 virus-neutralizing antibodies. Further, these neutralizing antibodies appear to be exclusively directed toward domain III of the DENV-3 E VLPs. Significantly, they also lack discernible ADE potential toward heterotypic DENVs. Taken together with the high productivity of the P. pastoris expression system, this approach could potentially pave the way toward developing a DENV E-based, inexpensive, safe, and efficacious tetravalent sub-unit vaccine, for use in resource-poor dengue endemic countries.
authors.affiliation: Recombinant Gene Products Group, International Centre for Genetic Engineering and Biotechnology, New Delhi India.
authors.forename: Lav
authors.initials: L
authors.lastname: Tripathi
authors.affiliation: Recombinant Gene Products Group, International Centre for Genetic Engineering and Biotechnology, New Delhi India.
authors.forename: Shailendra
authors.initials: S
authors.lastname: Mani
authors.affiliation: Recombinant Gene Products Group, International Centre for Genetic Engineering and Biotechnology, New Delhi India.
authors.forename: Rajendra
authors.initials: R
authors.lastname: Raut
authors.affiliation: Recombinant Gene Products Group, International Centre for Genetic Engineering and Biotechnology, New Delhi India.
authors.forename: Ankur
authors.initials: A
authors.lastname: Poddar
authors.affiliation: Recombinant Gene Products Group, International Centre for Genetic Engineering and Biotechnology, New Delhi India.
authors.forename: Poornima
authors.initials: P
authors.lastname: Tyagi
authors.affiliation: Recombinant Gene Products Group, International Centre for Genetic Engineering and Biotechnology, New Delhi India.
authors.forename: Upasana
authors.initials: U
authors.lastname: Arora
authors.affiliation: Department of Microbiology and Immunology, University of North Carolina School of Medicine, Chapel Hill, NC USA.
authors.forename: Aravinda
authors.initials: A
authors.lastname: de Silva
authors.affiliation: Department of Biological Sciences, Birla Institute of Technology and Sciences, Hyderabad India.
authors.forename: Sathyamangalam
authors.initials: S
authors.lastname: Swaminathan
authors.affiliation: Recombinant Gene Products Group, International Centre for Genetic Engineering and Biotechnology, New Delhi India ; Translational Health Science and Technology Institute, NCR Biotech Science Cluster, Faridabad India ; Department of Pediatrics, Emory University School of Medicine, Atlanta, GA USA.
authors.forename: Navin
authors.initials: N
authors.lastname: Khanna
keywords: Pichia pastoris; antibody-dependent enhancement (ADE); dengue; dengue envelope; dengue vaccine; neutralizing antibody; virus-like particle (VLP)