title: The application of an alanine-substituted mutant of the C-terminal fragment of Clostridium perfringens enterotoxin as a mucosal vaccine in mice.
abstract: Efficient delivery of antigen to mucosal immune tissues is an essential part of mucosal vaccination. Claudin-4 is expressed on the epithelial cells that cover the mucosal immune tissues. We previously found that claudin-4-targeting is a promising strategy for mucosal vaccination by using a claudin-4 binder, the C-terminal fragment of Clostridium perfringens enterotoxin (C-CPE). Substitution of Asn and Ser at positions 309 and 313, respectively, with alanine increased the affinity of C-CPE for claudin-4. However, application of the C-CPE mutant as a mucosal vaccine has never been tried. Here, we investigated whether the C-CPE mutant could serve as a mucosal vaccine. We used ovalbumin (OVA) as a model antigen and fused the C-CPE mutant to it. The resultant fusion protein was bound to claudin-4. When mice were immunized with the C-CPE mutant-fused OVA, OVA-specific serum IgG and nasal IgA increased relative to levels in mice immunized with a C-CPE-fused antigen. Immunization with the C-CPE mutant-fused OVA activated Th1- and Th2-type responses and led to increased anti-tumor activity against OVA-expressing thymoma cells relative to that of mice immunized with the C-CPE-fused antigen. These findings suggest that the alanine-substituted C-CPE mutant shows promise as a claudin-targeted mucosal vaccine.
authors: Laboratory of Bio-Functional Molecular Chemistry, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka 565-0871, Japan.
authors: Hidehiko
authors: H
authors: Suzuki
authors: Masuo
authors: M
authors: Kondoh
authors: Hideki
authors: H
authors: Kakutani
authors: Seiji
authors: S
authors: Yamane
authors: Hiroshi
authors: H
authors: Uchida
authors: Takao
authors: T
authors: Hamakubo
authors: Kiyohito
authors: K
authors: Yagi
mesh_terms: D000409:Alanine / Q000737:chemistry* / Q000235:genetics; D000818:Animals; D001428:Bacterial Vaccines / Q000235:genetics / Q000276:immunology*; D062506:Claudin-4; D057167:Claudins / Q000276:immunology* / Q000378:metabolism; D003015:Clostridium Infections / Q000276:immunology / Q000517:prevention & control; D003016:Clostridium perfringens / Q000276:immunology; D004768:Enterotoxins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D009092:Mucous Membrane / Q000276:immunology* / Q000378:metabolism / Q000382:microbiology; D010047:Ovalbumin / Q000276:immunology
chemical_list: D001428:Bacterial Vaccines; D062506:Claudin-4; D057167:Claudins; C568158:Cldn4 protein, mouse; D004768:Enterotoxins; C035598:enterotoxin, Clostridium; D010047:Ovalbumin; D000409:Alanine