Physicochemical and immunological characterization of Amb a 12, a novel ragweed (Ambrosia artemisiifolia) pollen allergen.

BACKGROUND

Ragweed is an invasive plant in Europe, causing hay fever and asthma in allergic patients. Climate change is predicted to increase expansion and allergenicity. Elevated NO induced upregulation of a new allergen in ragweed pollen, an enolase, Amb a 12.

OBJECTIVE

of this study was producing ragweed enolase as a recombinant protein and characterizing its physicochemical and immunological features.

METHODS

Amb a 12 was designed for E. coli and insect cell expression. Physicochemical features were determined by mass spectrometry, circular dichroism measurements and enzymatic activity assay. Immunological characteristics were determined in ELISA, in a mediator release assay and by investigation of association with clinical symptoms. Common allergen sources were screened for similar proteins.

RESULTS

Ragweed enolase was produced as a 48 kDa protein forming oligomers in both expression systems, showing differences in secondary structure content and enzymatic activity depending on expression system. IgE frequency and allergenicity were low regardless of expression system. Enolase-specific serum bound to similar sized molecules in mugwort, timothy grass and birch pollen, as well as food allergen sources, while highest IgE inhibition was achieved with peach pulp extract.

CONCLUSIONS

Amb a 12 had high sequence similarity and comparable IgE frequency to enolase allergens from different sources. 50 kDa proteins were found in other pollen and food allergen sources, suggesting that enolases might be pan-allergens in pollen and plant foods.