title: Antigen 43/Fcε3 chimeric protein expressed by a novel bacterial surface expression system as an effective asthma vaccine.
abstract: The IgE Fcε3 domain is an active immunotherapeutic target for asthma and other allergic diseases. However, previous methods for preparing IgE fusion protein vaccines are complex. Antigen 43 (Ag43) is a surface protein found in Escherichia coli that contains α and β subunits (the α subunit contains multiple T epitopes). Here we constructed a novel Ag43 surface display system (Ag43 system) to express Ag43 chimeric proteins to disrupt immune tolerance against IgE. The Ag43 system was constructed from the E. coli strain Tan109, in which the Ag43 gene was deleted and a recombinant plasmid (pETAg43) expressing a partial Ag43 gene was introduced. The Fcε3 domain of the IgE gene was then subcloned into plasmid pETAg43, resulting in a recombinant plasmid pETAg43/Fcε3, which was used to transform Tan109 for Ag43/Fcε3 surface expression. Thereafter, Ag43/Fcε3 was investigated as an asthma vaccine in a mouse model. Ag43/Fcε3 was expressed on and could be separated from the bacterial surface by heating to 60° while retaining activity. Ag43/Fcε3, as a protein vaccine, produced neutralizing autoantibodies to murine IgE, induced significant anti-asthma effects, and regulated IgE and T helper cytokines in a murine asthma model. Data show that Ag43/Fcε3 chimeric protein is a potential model vaccine for asthma treatment, and that the Ag43 system may be an effective tool for novel vaccine preparation to break immune tolerance to other self-molecules.
authors.affiliation: Hainan Provincial Key Laboratory of Tropical Medicine, Hainan Medical College, Haikou, China.
authors.forename: Feng-Ying
authors.initials: FY
authors.lastname: Huang
authors.forename: Cai-Chun
authors.initials: CC
authors.lastname: Wang
authors.forename: Yong-Hao
authors.initials: YH
authors.lastname: Huang
authors.forename: Huan-Ge
authors.initials: HG
authors.lastname: Zhao
authors.forename: Jun-Li
authors.initials: JL
authors.lastname: Guo
authors.forename: Song-Lin
authors.initials: SL
authors.lastname: Zhou
authors.forename: Hua
authors.initials: H
authors.lastname: Wang
authors.forename: Ying-Ying
authors.initials: YY
authors.lastname: Lin
authors.forename: Guang-Hong
authors.initials: GH
authors.lastname: Tan
mesh_terms: D018830:Adhesins, Escherichia coli / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D019264:Adoptive Transfer; D000818:Animals; D057134:Antibodies, Neutralizing / Q000097:blood; D001249:Asthma / Q000097:blood / Q000276:immunology / Q000503:physiopathology / Q000517:prevention & control*; D001323:Autoantibodies / Q000097:blood; D016535:Bronchial Hyperreactivity / Q000097:blood / Q000276:immunology / Q000503:physiopathology / Q000517:prevention & control*; D016084:Bronchoconstriction; D002478:Cells, Cultured; D003001:Cloning, Molecular; D016207:Cytokines / Q000378:metabolism; D004195:Disease Models, Animal; D006632:Histamine / Q000378:metabolism; D007108:Immune Tolerance; D007073:Immunoglobulin E / Q000276:immunology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010047:Ovalbumin / Q000276:immunology; D017455:Receptors, IgE / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology; D013997:Time Factors; D014614:Vaccines, Synthetic / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*
chemical_list: D018830:Adhesins, Escherichia coli; D057134:Antibodies, Neutralizing; D001323:Autoantibodies; D016207:Cytokines; D017455:Receptors, IgE; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; C118376:antigen 43, E coli; D007073:Immunoglobulin E; D006632:Histamine; D010047:Ovalbumin
keywords: IgE; antigen 43 (Ag43) surface display system; asthma; chronic airway hyper-reactivity; recombinant vaccine
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