Recombinant Immunoglobulin A Specific for Influenza A Virus Hemagglutinin: Production, Functional Analysis, and Formation of Secretory Immunoglobulin A

• Published in: Viral immunology Vol. 28; no. 3; pp. 17 - 178
• Main Authors: Shoji, Kentaro, Takahashi, Tadanobu, Kurohane, Kohta, Iwata, Koki, Matsuoka, Takeshi, Tsuruta, Shogo, Sugino, Takatomo, Miyake, Masaki, Suzuki, Takashi, Imai, Yasuyuki
• Format: Journal Article
• Language: English
• Published: United States Mary Ann Liebert, Inc 01.04.2015
• Subjects: Animals; Antibodies, Viral - genetics; Antibodies, Viral - immunology; Antibodies, Viral - isolation & purification; CHO Cells; Cricetulus; Dogs; Hemagglutinin Glycoproteins, Influenza Virus - immunology; Immunoglobulin A, Secretory - genetics; Immunoglobulin A, Secretory - immunology; Immunoglobulin A, Secretory - isolation & purification; Influenza A virus; Influenza A virus - immunology; Influenza A virus - physiology; Madin Darby Canine Kidney Cells; Mice; Original Research Articles; Original s; Recombinant Proteins - genetics; Recombinant Proteins - immunology; Recombinant Proteins - isolation & purification; Virus Internalization - drug effects
• ISSN: 0882-8245; 1557-8976
• DOI: 10.1089/vim.2014.0098

Secretory immunoglobulin (Ig) A (SIgA), comprised of dimeric IgA and secretory component (SC), is believed to provide a defense mechanism on the mucosal surface. Influenza A virus (IAV) hemagglutinin (HA)-specific SIgA is thought to play an important role in the prevention of IAV infection. However, the topical application of preformed IAV-specific SIgA has not been shown to prevent IAV infection. This is due to the difficulty in the production of antigen-specific IgA monoclonal antibodies (mAbs) and monoclonal SIgA. Here, a recombinant hybrid IgA (HIgA) was established that utilizes variable regions of an HA-specific mouse IgG mAb and the heavy chain constant region of a mouse IgA mAb. We expressed the dimeric HIgA in Chinese hamster ovary-K1 (CHO-K1) cells. When in vitro IAV infection of Madin–Darby canine kidney (MDCK) cells was tested, 10 times lower concentrations of HIgA were able to inhibit it as compared with an HA-specific IgG with the same variable regions. A functional hybrid secretory IgA (HSIgA) was also produced through incubation of the dimeric HIgA with recombinant mouse SC in vitro . It was demonstrated that HSIgA could be separated from the dimeric HIgA on size exclusion chromatography. This study provides a basic strategy for investigating the role of SIgA upon IAV infection on the mucosal surface.