title: Construction of a chimeric secretory IgA and its neutralization activity against avian influenza virus H5N1.
abstract: 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.
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Cun
authors.initials: C
authors.lastname: Li
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Xiaoping
authors.initials: X
authors.lastname: An
authors.affiliation: Centre of Excellence in Molecular Biology (CEMB), University of the Punjab, Lahore-53700, Pakistan.
authors.forename: Azeem Mehmood
authors.initials: AM
authors.lastname: Butt
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Baozhong
authors.initials: B
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Zhiyi
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Xiaona
authors.initials: X
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Huang
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Wenhui
authors.initials: W
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Mi
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Yigang
authors.initials: Y
authors.lastname: Tong
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing; D000941:Antigens / Q000276:immunology / Q000378:metabolism; D016466:CHO Cells; D003412:Cricetulus; D015870:Gene Expression; D023061:Gene Order; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007071:Immunoglobulin A, Secretory / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D053124:Influenza A Virus, H5N1 Subtype / Q000276:immunology*; D051379:Mice; D009500:Neutralization Tests; D009976:Orthomyxoviridae Infections / Q000276:immunology / Q000401:mortality / Q000821:virology; D011485:Protein Binding / Q000276:immunology; D011993:Recombinant Fusion Proteins
chemical_list: D057134:Antibodies, Neutralizing; D000941:Antigens; D007071:Immunoglobulin A, Secretory; D011993:Recombinant Fusion Proteins
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