Isolation and characterization of the major allergen from Italian cypress ( Cupressus sempervirens) pollen

• Published in: Journal of allergy and clinical immunology 2004-02, Vol.113 (2), p.S300-S300
• Main Authors: Togawa, A., Panzani, R.C., Garza, M., Goldblum, R.M., Midoro-Horiuti, T.
• Format: Article
• Language: English
• ISSN: 0091-6749; 1097-6825
• DOI: 10.1016/j.jaci.2004.01.562

The prevalence of seasonal allergic diseases of the upper airways is high in many industrialized countries and increasing. The family Cupressaceae is an important cause of pollinosis, particularly in Europe. Based on our previous studies of mountain cedar pollen, we hypothesized that the Italian cypress pollen would contain an allergenic, pathogenesis-related group 5 (PR-5) protein. We therefore sought to clone a cDNA from that pollen, based on nucleotide homology to Jun a 3 of mountain cedar. mRNA was purified from Italian cypress pollen. cDNA was synthesized and portion of a PR-5 message was amplified by PCR and extended by RACE methods. Recombinant Cup s 3 was expressed as a fusion protein with maltose binding protein. The allergenicity of Cup s 3 was established using an ELISA inhibition assay, based on crude extracts of Italian cedar pollen, the recombinant fusion protein of Cup s 3 and sera from France patients allergic to Italian cypress. Three isotypes, Cup s 3.1, Cup s 3.2 and Cup s 3.3. were identified. These had 95.5% amino acid sequence identity to Jun a 3. Recombinant Cup s 3.2 inhibited the binding of IgE from the majority of sera from patients to an extract of Italian cypress. Cup s 3 is a major allergen of Italian cypress. Anticipated variations in the content of Cup s 3 in pollen from different region/trees should be considered in the preparation of extracts for diagnostic and therapeutic use.