Isolation and characterization of native Cry j 3 from Japanese cedar (Cryptomeria japonica) pollen

• Published in: Allergy Vol. 62; no. 5; pp. 547 - 553
• Main Authors: Fujimura, T, Futamura, N, Midoro-Horiuti, T, Togawa, A, Goldblum, R.M, Yasueda, H, Saito, A, Shinohara, K, Masuda, K, Kurata, K, Sakaguchi, M
• Format: Journal Article
• Language: English
• Published: Oxford, UK Oxford, UK : Blackwell Publishing Ltd 01-05-2007; Blackwell Publishing Ltd; Blackwell
• Subjects: Allergic diseases; Amino Acid Sequence; Antigens, Plant - immunology; Antigens, Plant - isolation & purification; Antigens, Plant - metabolism; Base Sequence; Binding Sites, Antibody; Biological and medical sciences; Cryptomeria - chemistry; Cryptomeria - genetics; Cryptomeria - immunology; Cryptomeria japonica; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Humans; Hypersensitivity - immunology; Hypersensitivity - metabolism; Immunoglobulin E - biosynthesis; Immunoglobulin E - blood; Immunopathology; Japanese cedar; Juniperus ashei; Medical sciences; Molecular Sequence Data; oral allergy syndrome; pathogenesis-related protein; Pollen - chemistry; Pollen - immunology; pollinosis; Protein Binding - immunology; Protein Isoforms - immunology; Protein Isoforms - metabolism; thaumatin-like protein; Immunopathology; Thaumatin; Pathogenesis related protein; Japanese cedar; pollinosis; Crossed allergy; Immunology; Cryptomeria japonica; thaumatin-like protein; pathogenesis-related protein; Gymnospermae; Coniferales; Spermatophyta; Pollen; Oral allergy syndrome
• ISSN: 0105-4538; 1398-9995; 0108-1675
• DOI: 10.1111/j.1398-9995.2007.01331.x

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. 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. 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. Cry j 3 is a causative allergen in Japanese cedar pollinosis and may play crucial roles in the cross-reactivity with oral allergy syndrome.