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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...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2012
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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. |
format | Online Article Text |
id | pubmed-3511423 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
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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