Exhaustive Allergenome Analysis of Japanese Cedar Pollen Allergen Molecules

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Japanese cedar pollinosis is now a nation-wide allergic disease, although our knowledge about the causing factors, i.e. repertoire of Japanese cedar pollen (JCP) allergen molecules, is still limited. Objective of this study is exhaustive identification and characterization of JCP allergens via proteomics in conjunction with molecular immunological strategies, which we call "Allergenome Analysis".(1) Mapping of JCP allergensTwo-dimensional (2D) IgE immunoblotting revealed that; 1) total of 131 allergen spots were found on the 2D map, and major allergens Cry j 1 and Cry j 2 spots distributed as multiple isoforms. 2) IgE-binding profile of each pollinosis patients was quite different from each other, suggesting that tailor-made molecular diagnosis is crucial for effective immunotherapy. 3) 31 allergen spots showed higher IgE binding frequency than that of Cry j 2 (40%).(2) Identification and immunochemical characterization of new major JCP allergensWe next tried structural and immunochemical analyses of above new CJP allergens using TOF-MS analysis and cDNA cloning. We first identified class IV chitinase homologue (CJP4) as a novel major allergen with 100% IgE binding frequency, which is higher than that of Cry j 1 (71%). We also cloned two other allergens homologous to β-1,3-glucanase (CPA39) and aspartyl protease/nucleoid DNA binding protein family (CPA63), and functionally expressed those recombinant allergens using baculovirus-insect cell culture system. Intriguingly, we found that CJP4 and CPA39 showed IgE crossreactivity with other plant species such as latex or olive pollen, suggesting that JCP chitinase and/or β-1,3-glucanase may serve as pan-allergens involved in the pathogenesis of oral allergy syndrome.