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Isolation and characterization of native Cry j 3 from Japanese cedar ( Cryptomeria japonica ) pollen
Author(s) -
Fujimura T.,
Futamura N.,
MidoroHoriuti T.,
Togawa A.,
Goldblum R. M.,
Yasueda H.,
Saito A.,
Shinohara K.,
Masuda K.,
Kurata K.,
Sakaguchi M.
Publication year - 2007
Publication title -
allergy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.363
H-Index - 173
eISSN - 1398-9995
pISSN - 0105-4538
DOI - 10.1111/j.1398-9995.2007.01331.x
Subject(s) - cryptomeria , allergen , immunoglobulin e , pollen , japonica , complementary dna , histamine , biology , allergy , antibody , immunology , cdna library , microbiology and biotechnology , botany , biochemistry , gene , endocrinology
Background: Japanese cedar ( Cryptomeria japonica ) pollinosis is the most prevalent allergy in Japan. Recently, the Japanese cedar pollen allergen Cry j 3 was cloned as a homologue of Jun a 3, which is a major allergen from mountain cedar ( Juniperus ashei ) pollen. However, native Cry j 3 has not been isolated and there are no reports on its allergenic activity. The aims of this study were to isolate native Cry j 3 and assess its immunoglobulin E (IgE)‐binding capacity in patients with Japanese cedar pollinosis. Methods: Native Cry j 3 was purified from Japanese cedar pollen by multidimensional chromatography. We assessed the IgE‐binding capacity using sera from patients allergic to Japanese cedar pollen by immunoblot analysis and ELISA. Moreover, we assayed the capacity of Cry j 3 to induce histamine release from the patients’ leukocytes. We cloned cDNA corresponding to purified Cry j 3 from a cDNA library of Japanese cedar pollen. Results: We isolated native Cry j 3 as a 27‐kDa protein. The IgE‐binding frequency of Cry j 3 from the sera of patients allergic to Japanese cedar pollen was estimated as 27% (27/100) by ELISA. Cry j 3 induced the release of histamine from leukocytes. We cloned the cDNA and named it Cry j 3.8. Cry j 3.8 cDNA encoded 225 amino acids and had significant homology with thaumatin‐like proteins. Conclusions: Cry j 3 is a causative allergen in Japanese cedar pollinosis and may play crucial roles in the cross‐reactivity with oral allergy syndrome.