Identification of Distinctive Variants of the Olive Pollen Allergen Ole e 5 (Cu,Zn Superoxide Dismutase) throughout the Analysis of the Olive Pollen Transcriptome

摘要

Ole e 5 is an olive pollen allergen displaying high identity with Cu,Zn superoxide dismutases. Previous studies characterized biochemical variability in this allergen, which may be of relevance for allergy diagnosis and therapy. The generation of an olive pollen transcriptome allowed us to identify eight Ole e 5 sequences, one of them including a 24 nt deletion. Further in silico analysis permitted designing primers for PCR amplification and cloning from both cDNA and gDNA. A large number of sequences were retrieved, which experimentally validated the predictive NGS sequences, including the deleted enzyme. The PCR-obtained sequences were used for further scrutiny, including sequence aligment and phylogenetic analysis. Two model sequences (a complete sequence and a deleted one) were used to perform 3-D modeling and a prediction of the T- and B-cell epitopes. These predictions interestingly foreseed relevant differences in the antigenicity/allergenicity of both molecules. Clinical relevance of differences is discussed.