Comparison of Four Grass Pollen Species Concerning Their Allergens of Grass Group V by 2D Immunoblotting and Microsequencing

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Abstract
The identification and characterization of allergenic components is a vital step towards improving diagnosis and therapy. Members of the grass family (Poaceae) reveal a high cross-reactivity among each other caused by the close phylogenetical relationship. In order to investigate the variability between allergenic components, we studied the allergen grass group V, one of the major allergens. Pollen extracts of 4 different tribes (timothy grass (Phleum pratense)--Agrostidae, perennial rye grass (Lolium perenne)--Festuceae, meadow velvet (Holcus lanatus)--Aveneae, and rye (Secale cereale)--Triticeae) of the Festucoideae subfamily were separated by 2D PAGE and investigated by immunoblotting using patients' poolserum and monoclonal antibodies (raised against group V allergens of timothy grass pollen). The antibodies identify different allergens in the four grass species. The components vary from 30-50 kDa and pI 4.8-7.0. The eight NH2-terminal amino acids were determined and indicated high similarities between the different components. These results cast doubt on the suitability of classifying allergens into groups based only on their molecular mass, isoelectric point and N-terminal sequence analysis. It suggests to classify allergens according to their IgE-reactive epitopes.