Epitope analysis of Japanese cedar pollen allergen Cry j2 with a human IgM class monoclonal antibody 404-117

Article type: Research Article

Authors: Chiba, Seiya^a | Yokoyama, Haruka^a | Kumazawa, Ayane^a | Shimmoto, Michie^b | Naganawa, Yasunori^a | Shinmoto, Hiroshi^{a; *}

Affiliations: [a] Tamagawa University, College of Agriculture, Machida, Tokyo, Japan | [b] Tokyo Metropolitan Institute of Medical Science, Setagaya-ku, Tokyo, Japan

Correspondence: [*] Corresponding author: Hiroshi Shinmoto, College of Agriculture, Tamagawa University, 6-1-1 Tamagawa-gakuen, Machida, Tokyo 194-8610, Japan. Tel./Fax: +81 42 739 8267; E-mail:[email protected]

Abstract: Japanese cedar pollen allergen Cry j2 is a causal allergen of seasonal pollinosis in Japan. To analyze B cell epitopes of Cry j2, we established two human-mouse hybridomas secreting IgM class human monoclonal antibodies to Cry j2. A pin-peptide enzyme-linked immunosorbent assay with synthesized icosa peptides showed that 404-117 monoclonal antibody bound to peptides #11-13 with cry j2 amino acid sequence of 101F-L140. Detailed analysis with octa peptides and alanine substituted peptides indicated that an amino acid sequence of 118FKVD121 was an essential for antibody binding. When K119 (Asn) was substituted with alanine, 404-117 monoclonal antibody did not bind to the alanine substituted peptide. We concluded that the 118FKVD121 sequence might have a very important role in early recognition by Cry j2-specific B cells, which could act as antigen presenting cells.

Keywords: Human monoclonal antibody, allergen, Japanese cedar pollen, epitope, IgM

DOI: 10.3233/HAB-160301

Journal: Human Antibodies, vol. 25, no. 1-2, pp. 17-21, 2017