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Immunological and Biochemical Characterization of Coxsackie Virus A16 Viral Particles

BACKGROUND: Coxsackie virus A16 (CVA16) infections have become a serious public health problem in the Asia-Pacific region. It manifests most often in childhood exanthema, commonly known as hand-foot-and-mouth disease (HFMD). There are currently no vaccine or effective medical treatments available. P...

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Autores principales: Chong, Pele, Guo, Meng-Shin, Lin, Fion Hsiao-Yu, Hsiao, Kuang-Nan, Weng, Shu-Yang, Chou, Ai-Hsiang, Wang, Jen-Ren, Hsieh, Shih-Yang, Su, Ih-Jen, Liu, Chia-Chyi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3511423/
https://www.ncbi.nlm.nih.gov/pubmed/23226233
http://dx.doi.org/10.1371/journal.pone.0049973
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author Chong, Pele
Guo, Meng-Shin
Lin, Fion Hsiao-Yu
Hsiao, Kuang-Nan
Weng, Shu-Yang
Chou, Ai-Hsiang
Wang, Jen-Ren
Hsieh, Shih-Yang
Su, Ih-Jen
Liu, Chia-Chyi
author_facet Chong, Pele
Guo, Meng-Shin
Lin, Fion Hsiao-Yu
Hsiao, Kuang-Nan
Weng, Shu-Yang
Chou, Ai-Hsiang
Wang, Jen-Ren
Hsieh, Shih-Yang
Su, Ih-Jen
Liu, Chia-Chyi
author_sort Chong, Pele
collection PubMed
description BACKGROUND: Coxsackie virus A16 (CVA16) infections have become a serious public health problem in the Asia-Pacific region. It manifests most often in childhood exanthema, commonly known as hand-foot-and-mouth disease (HFMD). There are currently no vaccine or effective medical treatments available. PRINCIPAL FINDING: In this study, we describe the production, purification and characterization of CVA16 virus produced from Vero cells grown on 5 g/L Cytodex 1 microcarrier beads in a five-liter serum-free bioreactor system. The viral titer was found to be >10(6) the tissue culture's infectious dose (TCID(50)) per mL within 7 days post-infection when a multiplicity of infection (MOI) of 10(−5) was used for initial infection. Two CVA16 virus fractions were separated and detected when the harvested CVA16 viral concentrate was purified by a sucrose gradient zonal ultracentrifugation. The viral particles detected in the 24–28% sucrose fractions had low viral infectivity and RNA content. The viral particles obtained from 35–38% sucrose fractions were found to have high viral infectivity and RNA content, and composed of four viral proteins (VP1, VP2, VP3 and VP4), as shown by SDS-PAGE analyses. These two virus fractions were formalin-inactivated and only the infectious particle fraction was found to be capable of inducing CVA16-specific neutralizing antibody responses in both mouse and rabbit immunogenicity studies. But these antisera failed to neutralize enterovirus 71. In addition, rabbit antisera did not react with any peptides derived from CVA16 capsid proteins. Mouse antisera recognized a single linear immunodominant epitope of VP3 corresponding to residues 176–190. CONCLUSION: These results provide important information for cell-based CVA16 vaccine development. To eliminate HFMD, a bivalent EV71/CVA16 vaccine formulation is necessary.
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spelling pubmed-35114232012-12-05 Immunological and Biochemical Characterization of Coxsackie Virus A16 Viral Particles Chong, Pele Guo, Meng-Shin Lin, Fion Hsiao-Yu Hsiao, Kuang-Nan Weng, Shu-Yang Chou, Ai-Hsiang Wang, Jen-Ren Hsieh, Shih-Yang Su, Ih-Jen Liu, Chia-Chyi PLoS One Research Article BACKGROUND: Coxsackie virus A16 (CVA16) infections have become a serious public health problem in the Asia-Pacific region. It manifests most often in childhood exanthema, commonly known as hand-foot-and-mouth disease (HFMD). There are currently no vaccine or effective medical treatments available. PRINCIPAL FINDING: In this study, we describe the production, purification and characterization of CVA16 virus produced from Vero cells grown on 5 g/L Cytodex 1 microcarrier beads in a five-liter serum-free bioreactor system. The viral titer was found to be >10(6) the tissue culture's infectious dose (TCID(50)) per mL within 7 days post-infection when a multiplicity of infection (MOI) of 10(−5) was used for initial infection. Two CVA16 virus fractions were separated and detected when the harvested CVA16 viral concentrate was purified by a sucrose gradient zonal ultracentrifugation. The viral particles detected in the 24–28% sucrose fractions had low viral infectivity and RNA content. The viral particles obtained from 35–38% sucrose fractions were found to have high viral infectivity and RNA content, and composed of four viral proteins (VP1, VP2, VP3 and VP4), as shown by SDS-PAGE analyses. These two virus fractions were formalin-inactivated and only the infectious particle fraction was found to be capable of inducing CVA16-specific neutralizing antibody responses in both mouse and rabbit immunogenicity studies. But these antisera failed to neutralize enterovirus 71. In addition, rabbit antisera did not react with any peptides derived from CVA16 capsid proteins. Mouse antisera recognized a single linear immunodominant epitope of VP3 corresponding to residues 176–190. CONCLUSION: These results provide important information for cell-based CVA16 vaccine development. To eliminate HFMD, a bivalent EV71/CVA16 vaccine formulation is necessary. Public Library of Science 2012-11-30 /pmc/articles/PMC3511423/ /pubmed/23226233 http://dx.doi.org/10.1371/journal.pone.0049973 Text en © 2012 Chong et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Chong, Pele
Guo, Meng-Shin
Lin, Fion Hsiao-Yu
Hsiao, Kuang-Nan
Weng, Shu-Yang
Chou, Ai-Hsiang
Wang, Jen-Ren
Hsieh, Shih-Yang
Su, Ih-Jen
Liu, Chia-Chyi
Immunological and Biochemical Characterization of Coxsackie Virus A16 Viral Particles
title Immunological and Biochemical Characterization of Coxsackie Virus A16 Viral Particles
title_full Immunological and Biochemical Characterization of Coxsackie Virus A16 Viral Particles
title_fullStr Immunological and Biochemical Characterization of Coxsackie Virus A16 Viral Particles
title_full_unstemmed Immunological and Biochemical Characterization of Coxsackie Virus A16 Viral Particles
title_short Immunological and Biochemical Characterization of Coxsackie Virus A16 Viral Particles
title_sort immunological and biochemical characterization of coxsackie virus a16 viral particles
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3511423/
https://www.ncbi.nlm.nih.gov/pubmed/23226233
http://dx.doi.org/10.1371/journal.pone.0049973
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