title: Recombinant immunoglobulin A specific for influenza A virus hemagglutinin: production, functional analysis, and formation of secretory immunoglobulin A.
abstract: 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.
authors: 1 Laboratory of Microbiology and Immunology, School of Pharmaceutical Sciences, University of Shizuoka , Shizuoka, Japan .
authors: Kentaro
authors: K
authors: Shoji
authors: Tadanobu
authors: T
authors: Takahashi
authors: Kohta
authors: K
authors: Kurohane
authors: Koki
authors: K
authors: Iwata
authors: Takeshi
authors: T
authors: Matsuoka
authors: Shogo
authors: S
authors: Tsuruta
authors: Takatomo
authors: T
authors: Sugino
authors: Masaki
authors: M
authors: Miyake
authors: Takashi
authors: T
authors: Suzuki
authors: Yasuyuki
authors: Y
authors: Imai
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D016466:CHO Cells; D003412:Cricetulus; D004285:Dogs; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000276:immunology*; D007071:Immunoglobulin A, Secretory / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D009980:Influenza A virus / Q000276:immunology* / Q000502:physiology; D061985:Madin Darby Canine Kidney Cells; D051379:Mice; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D053586:Virus Internalization / Q000187:drug effects
chemical_list: D000914:Antibodies, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D007071:Immunoglobulin A, Secretory; D011994:Recombinant Proteins