Author/Authors :
Varasteh، Abdol-Reza نويسنده Allergy Research Center, Mashhad University of Medical Sciences, Mashhad, Iran , , Sankian، Mojtaba نويسنده , , Midoro-Horiuti، Terumi نويسنده INDOOR Biotechnologies, Inc., Charlottesville, Virginia, USA , , Moghadam، Malihe نويسنده Immuno-Biochemistry Lab, Immunology Research Center, Mashhad University of Medical Sciences (MUMS), Mashhad, Iran , , Shakeri، Mohamad Taghi نويسنده Immuno-Biochemistry Lab, Immunology Research Center, Mashhad University of Medical Sciences (MUMS), Mashhad, Iran , , Brooks، Edward G. نويسنده , , Goldblum، Randall M نويسنده Department of Pediatrics, University of Texas Medical Branch, Galveston, Texas, USA , , Chapman، Martin C. نويسنده , , Pomés، Anna نويسنده INDOOR Biotechnologies, Inc., Charlottesville, Virginia, USA. ,
Abstract :
Background: The cultivation of saffron is expanding through the southeast of Iran, and allergy to
saffron pollen occurs in workers involved in processing this plant. We aimed to clone, sequence and
express a major allergen involved in saffron pollen allergy, and to compare the recombinant with the
natural allergen.
Methods: The N-terminal amino acid sequence of Cro s 1, an allergen from saffron pollen, was
determined after immunoblotting. The cDNA encoding for this allergen was cloned by PCR utilizing a
primer based on the N-terminal amino acid sequence. Recombinant Cro s 1 (rCro s 1) was expressed as
a soluble protein in Pichia pastoris and purified to homogeneity by gel filtration. Inhibition of IgE
binding to rCro s 1 by pollen extract was analyzed by ELISA.
Results: The allergen Cro s 1 was identified from saffron pollen extracts and cloned by PCR. Cro s 1
cDNA defined an acidic polypeptide with homology to pollen proteins from Chenopodium album and
Ligastrum vulgaris. The rCro s 1 was expressed in P. pastoris at 28 mg/l. Saffron pollen extract
inhibited the binding of patient serum IgE to rCro s 1.
Conclusion: We identified and cloned the first Crocus sativus pollen allergen. rCro s 1 cDNA shows a
very high homology with Che a 1, the major allergen of lambʹs-quarter, Chenopodium album,
Caryophyllales, pollen (97%). Cro s 1 is a useful tool for specific diagnosis and structural studies of
occupational allergy to saffron.
