Molecular characterization of a cross-reactive Juniperus oxycedrus pollen allergen, Jun o 2:: A novel calcium-binding allergen

JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY

卷:101

Article

Tinghino, R ; Barletta, B ; Palumbo, S ; Afferni, C ; Iacovacci, P ; Mari, A ; Di Felice, G ; Pini, C

关键词: Cupressaceae;    Juniperus oxycedrus;    cDNA cloning;    IgE;    calcium-binding protein;    cross reactivity;   

DOI  :  10.1016/S0091-6749(98)70306-9

来源: Elsevier

【 摘 要 】

Background: Species belonging to the Cupressaceae family are a relevant source of allergens that are present in a wide number of countries. Objective:We sought to identify, purify, and characterize recombinant allergens from Juniperus oxycedrus, a species belonging to the Cupressaceae family. Methods: Double-stranded cDNA was synthesized from mRNA and cloned into the lambda-ZAP expression vector. IgE screening of the Library was performed with a pool of sera from subjects allergic to Cupressaceae. A recombinant 6 x His-tagged Juniperus oxycedrus allergen, Jun o 2, was expressed in Escherichia coli and purified by Ni(2+) affinity chromatography. It was studied further by immunoblotting inhibition with pollen extracts from other Cupressaceae, Oleaceae, Urticaceae, and Graminaceae. The role of protein-bound calcium on the allergen's IgE-binding capacity was tested in a plaque assay in the presence or absence of EGTA. Results: A cDNA coding for a newly identified Juniperus oxycedrus pollen allergen, dun o 2, was isolated. The deduced amino acid sequence contained four typical Ca(2+) binding sites and showed a significant sequence similarity to calmodulins. Depletion of Ca(2+) in the plaque assay led to a loss of IgE-binding capacity of dun o 2. Immunoblotting inhibition revealed that J. oxycedrus, J. ashei, Cupressus arizonica, C. sempervirens, Parietaria judaica, Olea europaea, and Lolium perenne pollen extracts were able to inhibit IgE binding to blotted rJun o 2 at different concentrations. Conclusion: rJun o 2 contains IgE-binding epitopes shared by taxonomically unrelated species, and therefore it can be regarded as a new panallergen. These findings could contribute to an explanation for the phenomenon of multiple positive test results in polysensitized patients and the potential symptom-eliciting role of allergenic sources previously not encountered.

【 授权许可】

Free