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Development of novel claudin‐4 binder and its application in mucosal vaccine
Author(s) -
Yagi Kiyohito,
Yamane Seiji,
Suzuki Hidehiko,
Watari Akihiro,
Kondoh Masuo,
Uchida Hiroshi
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.lb389
Subject(s) - ovalbumin , antigen , enterotoxin , immune system , vaccination , mutant , immunization , chemistry , microbiology and biotechnology , immunology , medicine , biology , escherichia coli , biochemistry , gene
Mucosal vaccines activate both mucosal and systemic immune responses, but mucosal administration is needle‐free and less painful. Thus, mucosal vaccination appears to be an ideal vaccination strategy. The effective delivery of antigens to mucosa‐associated lymphoid tissues (MALT) is critical to the development of potent mucosal vaccines. We previously found that claudin‐4‐targeting is a promising strategy for mucosal vaccination using a claudin‐4 ligand, the C‐terminal fragment of Clostridium perfringens enterotoxin (C‐CPE). In the present study, we identified the functional domain of C‐CPE, and we attempted to prepare a novel claudin binder with higher affinity for claudin using the functional mapping information of C‐CPE. Substitution of some amino acids with alanine increased affinity of C‐CPE to claudin‐4, and Kd values decreased from 455 pM to 46 pM. To investigate mucosal vaccination using the mutant C‐CPE, ovalbumin (OVA) was used as a model antigen. When mice were immunized with the C‐CPE mutant‐fused antigen, antigen‐specific serum IgG and nasal IgA increased 10‐fold when compared with mice immunized using C‐CPE‐fused antigen. Immunization with the C‐CPE mutant‐fused antigen OVA activated Th1‐ and Th2‐type responses more strongly than in mice immunized with C‐CPE‐fused antigen. These results indicate that the alanine‐substituted C‐CPE mutant is a promising lead for claudin‐targeted mucosal vaccines.