title: Phage Display-Derived Ligand for Mucosal Transcytotic Receptor GP-2 Promotes Antigen Delivery to M Cells and Induces Antigen-Specific Immune Response.
abstract: Successful oral immunization depends on efficient delivery of antigens (Ags) to the mucosal immune induction site. Glycoprotein-2 (GP-2) is an integral membrane protein that is expressed specifically on M cells within follicle-associated epithelium (FAE) and serves as transcytotic receptor for luminal Ags. In this study, we selected peptide ligands against recombinant human GP-2 by screening a phage display library and evaluated their interaction with GP-2 in vitro and ex vivo. Selected peptides were conjugated to the C-terminal of enhanced green fluorescence protein (EGFP) and evaluated for their ability to induce an immune response in mice. One of our selected peptides, Gb-1, showed high binding affinity to GP-2 and, when fused to EGFP, significantly increased the uptake of EGFP by M cells compared to EGFP alone. After oral administration, the Gb1-EGFP fusion induced efficient mucosal and systemic immune responses in mice measured at the level of antigen-specific serum and fecal antibodies, cytokine secretion, and lymphocyte proliferation. Furthermore, the IgG subclasses and cytokine secretion showed that ligand Gb-1 induced a Th2-type immune response. Collectively, our findings suggest that the ligand we selected through phage library screening is capable of targeting Ags to GP-2 on M cells and can be used as an oral vaccine adjuvant.
authors.affiliation: 1 Laboratory of Molecular Immunology, State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai 200433, PR China.
authors.forename: Inam Ullah
authors.initials: IU
authors.lastname: Khan
authors.affiliation: 1 Laboratory of Molecular Immunology, State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai 200433, PR China.
authors.forename: Jiansheng
authors.initials: J
authors.lastname: Huang
authors.affiliation: 1 Laboratory of Molecular Immunology, State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai 200433, PR China.
authors.forename: Rui
authors.initials: R
authors.lastname: Liu
authors.affiliation: 1 Laboratory of Molecular Immunology, State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai 200433, PR China.
authors.forename: Jingbo
authors.initials: J
authors.lastname: Wang
authors.affiliation: 1 Laboratory of Molecular Immunology, State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai 200433, PR China.
authors.forename: Jun
authors.initials: J
authors.lastname: Xie
authors.affiliation: 1 Laboratory of Molecular Immunology, State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai 200433, PR China.
authors.forename: Naishuo
authors.initials: N
authors.lastname: Zhu
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000284:Administration, Oral; D000818:Animals; D000941:Antigens / Q000276:immunology*; D001435:Bacteriophages / Q000276:immunology*; D049109:Cell Proliferation / Q000502:physiology; D061505:Cell Surface Display Techniques / Q000379:methods; D004848:Epithelium / Q000276:immunology; D005260:Female; D049452:Green Fluorescent Proteins / Q000276:immunology; D006801:Humans; D018928:Immunity, Mucosal / Q000276:immunology*; D007114:Immunization / Q000379:methods; D008024:Ligands; D008214:Lymphocytes / Q000276:immunology; D008562:Membrane Glycoproteins / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009092:Mucous Membrane / Q000276:immunology; D010455:Peptides / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000276:immunology; D057900:Transcytosis / Q000276:immunology*; D014611:Vaccination / Q000379:methods
chemical_list: D000276:Adjuvants, Immunologic; D000941:Antigens; D008024:Ligands; D008562:Membrane Glycoproteins; D010455:Peptides; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
keywords: M cells; Peyer’s patches; adjuvant; glycoprotein-2; transcytosis