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Construction of a Chimeric Secretory IgA and Its Neutralization Activity against Avian Influenza Virus H5N1

Secretory immunoglobulin A (SIgA) acts as the first line of defense against respiratory pathogens. In this assay, the variable regions of heavy chain (VH) and Light chain (VL) genes from a mouse monoclonal antibody against H5N1 were cloned and fused with human IgA constant regions. The full-length c...

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Autores principales: Li, Cun, An, Xiaoping, Butt, Azeem Mehmood, Zhang, Baozhong, Zhang, Zhiyi, Wang, Xiaona, Huang, Yong, Zhang, Wenhui, Zhang, Bo, Mi, Zhiqiang, Tong, Yigang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3987799/
https://www.ncbi.nlm.nih.gov/pubmed/24741594
http://dx.doi.org/10.1155/2014/394127
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author Li, Cun
An, Xiaoping
Butt, Azeem Mehmood
Zhang, Baozhong
Zhang, Zhiyi
Wang, Xiaona
Huang, Yong
Zhang, Wenhui
Zhang, Bo
Mi, Zhiqiang
Tong, Yigang
author_facet Li, Cun
An, Xiaoping
Butt, Azeem Mehmood
Zhang, Baozhong
Zhang, Zhiyi
Wang, Xiaona
Huang, Yong
Zhang, Wenhui
Zhang, Bo
Mi, Zhiqiang
Tong, Yigang
author_sort Li, Cun
collection PubMed
description Secretory immunoglobulin A (SIgA) acts as the first line of defense against respiratory pathogens. In this assay, the variable regions of heavy chain (VH) and Light chain (VL) genes from a mouse monoclonal antibody against H5N1 were cloned and fused with human IgA constant regions. The full-length chimeric light and heavy chains were inserted into a eukaryotic expressing vector and then transfected into CHO/dhfr-cells. The chimeric monomeric IgA antibody expression was confirmed by using ELISA, SDS-PAGE, and Western blot. In order to obtain a dimeric secretory IgA, another two expressing plasmids, namely, pcDNA4/His A-IgJ and pcDNA4/His A-SC, were cotransfected into the CHO/dhfr-cells. The expression of dimeric SIgA was confirmed by using ELISA assay and native gel electrophoresis. In microneutralization assay on 96-well immunoplate, the chimeric SIgA showed neutralization activity against H5N1 virus on MDCK cells and the titer was determined to be 1 : 64. On preadministrating intranasally, the chimeric SIgA could prevent mice from lethal attack by using A/Vietnam/1194/04 H5N1 with a survival rate of 80%. So we concluded that the constructed recombinant chimeric SIgA has a neutralization capability targeting avian influenza virus H5N1 infection in vitro and in vivo.
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spelling pubmed-39877992014-04-16 Construction of a Chimeric Secretory IgA and Its Neutralization Activity against Avian Influenza Virus H5N1 Li, Cun An, Xiaoping Butt, Azeem Mehmood Zhang, Baozhong Zhang, Zhiyi Wang, Xiaona Huang, Yong Zhang, Wenhui Zhang, Bo Mi, Zhiqiang Tong, Yigang J Immunol Res Research Article Secretory immunoglobulin A (SIgA) acts as the first line of defense against respiratory pathogens. In this assay, the variable regions of heavy chain (VH) and Light chain (VL) genes from a mouse monoclonal antibody against H5N1 were cloned and fused with human IgA constant regions. The full-length chimeric light and heavy chains were inserted into a eukaryotic expressing vector and then transfected into CHO/dhfr-cells. The chimeric monomeric IgA antibody expression was confirmed by using ELISA, SDS-PAGE, and Western blot. In order to obtain a dimeric secretory IgA, another two expressing plasmids, namely, pcDNA4/His A-IgJ and pcDNA4/His A-SC, were cotransfected into the CHO/dhfr-cells. The expression of dimeric SIgA was confirmed by using ELISA assay and native gel electrophoresis. In microneutralization assay on 96-well immunoplate, the chimeric SIgA showed neutralization activity against H5N1 virus on MDCK cells and the titer was determined to be 1 : 64. On preadministrating intranasally, the chimeric SIgA could prevent mice from lethal attack by using A/Vietnam/1194/04 H5N1 with a survival rate of 80%. So we concluded that the constructed recombinant chimeric SIgA has a neutralization capability targeting avian influenza virus H5N1 infection in vitro and in vivo. Hindawi Publishing Corporation 2014 2014-02-13 /pmc/articles/PMC3987799/ /pubmed/24741594 http://dx.doi.org/10.1155/2014/394127 Text en Copyright © 2014 Cun Li et al. https://creativecommons.org/licenses/by/3.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Li, Cun
An, Xiaoping
Butt, Azeem Mehmood
Zhang, Baozhong
Zhang, Zhiyi
Wang, Xiaona
Huang, Yong
Zhang, Wenhui
Zhang, Bo
Mi, Zhiqiang
Tong, Yigang
Construction of a Chimeric Secretory IgA and Its Neutralization Activity against Avian Influenza Virus H5N1
title Construction of a Chimeric Secretory IgA and Its Neutralization Activity against Avian Influenza Virus H5N1
title_full Construction of a Chimeric Secretory IgA and Its Neutralization Activity against Avian Influenza Virus H5N1
title_fullStr Construction of a Chimeric Secretory IgA and Its Neutralization Activity against Avian Influenza Virus H5N1
title_full_unstemmed Construction of a Chimeric Secretory IgA and Its Neutralization Activity against Avian Influenza Virus H5N1
title_short Construction of a Chimeric Secretory IgA and Its Neutralization Activity against Avian Influenza Virus H5N1
title_sort construction of a chimeric secretory iga and its neutralization activity against avian influenza virus h5n1
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3987799/
https://www.ncbi.nlm.nih.gov/pubmed/24741594
http://dx.doi.org/10.1155/2014/394127
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