Please use this identifier to cite or link to this item: https://doi.org/10.1016/j.virol.2004.11.022
Title: West Nile premembrane-envelope genetic vaccine encoded as a chimera containing the transmembrane and cytoplasmic domains of a lysosome-associated membrane protein: Increased cellular concentration of the transgene product, targeting to the MHC II compartment, and enhanced neutralizing antibody response
Authors: Anwar, A.
Chandrasekaran, A.
Ng, M.L. 
Marques, E.
August, J.T.
Keywords: Electron microscopy
H-2M
LAMP
MHC II
MIICs
Neutralizing antibody
West Nile virus DNA vaccine
Issue Date: 5-Feb-2005
Citation: Anwar, A., Chandrasekaran, A., Ng, M.L., Marques, E., August, J.T. (2005-02-05). West Nile premembrane-envelope genetic vaccine encoded as a chimera containing the transmembrane and cytoplasmic domains of a lysosome-associated membrane protein: Increased cellular concentration of the transgene product, targeting to the MHC II compartment, and enhanced neutralizing antibody response. Virology 332 (1) : 66-77. ScholarBank@NUS Repository. https://doi.org/10.1016/j.virol.2004.11.022
Abstract: A genetic vaccine for West Nile virus (WN) has been synthesized with the WN premembrane-envelope (WN preM-E) gene sequences encoded as a chimera with the transmembrane and carboxyl terminal domains of the lysosome-associated membrane protein (LAMP). The LAMP sequences are used to direct the antigen protein to the major histocompatibility class II (MHC II) vesicular compartment of transfected professional antigen-presenting cells (APCs). Vaccine constructs encoding the native WN preM-E and WN preM-E/LAMP chimera were synthesized in pVAX1 and pITR plasmid backbones. Extracts of human fibroblast 293 and monkey kidney COS-7 cells transfected with the WN preM-E/LAMP chimera constructs contained much greater amounts of E than did the cells transfected with constructs encoding the native WN preM-E. This difference in the concentration of native E and the E/LAMP chimera in transfected cells is attributed to the secretion of native E. The amount of preM protein in cell extracts, in contrast to the E protein, and the levels of DNA and RNA transcripts, did not differ between WN preM-E- and WN preM-E/LAMP-transfected cells. Additionally, confocal and immunoelectron microscopic analyses of transfected B cells showed localization of the WN preM-E/LAMP chimera in vesicular compartments containing endogenous LAMP, MHC II, and H2-M, whereas native viral preM-E lacking the LAMP sequences was distributed within the cellular vesicular network with little LAMP or MHC II association. Mice immunized with a DNA construct expressing the WN preM-E/LAMP antigen induced significant antibody and long-term neutralization titers in contrast to the minimal and short-lived neutralization titer of mice vaccinated with a plasmid expressing the untargeted antigen. These results underscore the utility of LAMP targeting of the WN envelope to the MHC II compartments in the design of a genetic WN vaccine. © 2004 Elsevier Inc. All rights reserved.
Source Title: Virology
URI: http://scholarbank.nus.edu.sg/handle/10635/130350
ISSN: 00426822
DOI: 10.1016/j.virol.2004.11.022
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