Cross-Reactivity Among Pho d 2 and Profilins from Different Pollen Allergenic Sources

Abstract

RationaleProfilins are considered as an important panallergen being the IgE cross-reactivity among these proteins associated with multiple pollen sensitization. Sequence homology among pollen profilins is very high, ranging 67.4-89.5%. The objective was to evaluate the cross-reactivity among profilin from palm tree pollen (Phoenix dactylifera) and other pollen allergenic sources where profilin has been described.MethodsPalm tree profilin (Pho d 2) was purified by affinity chromatography in a poly-L-proline column, sequenced by LC/MS-MS and used to produce rabbit polyclonal antibodies. Identification of profilin in eleven pollen extracts (Ambrosia artemisiifolia, Artemisia vulgaris, Betula alba, Chenopodium album, Cynodon dactylon, Helianthus annus, Mercurialis perennis, Olea europaea, Parietaria judaica, Phleum pratense and Plantago lanceolata) was carried out by IgG ELISA and immunoblot using the polyclonal antibodies. Cross-reactivity studies were performed by IgG immunoblot inhibition assays, using the purified profilin for inhibition.ResultsHighly purified profilin (>95%) from palm tree pollen (Pho d 2) with a molecular weight of 14 kDa was obtained. Identity and purity of profilin was confirmed by LC/MS-MS. Presence of profilin was confirmed in all the extracts, except Chenopodium, by direct ELISA (O.D.>0.1). Identification of profilin by IgG immunoblot, and cross-reactivity with Pho d 2 by immunoblot-inhibition, was observed in Ambrosia, Betula, Cynodon, Helianthus, Mercurialis, Plantago and Phleum extracts.ConclusionsCross-reactivity among Pho d 2 and other profilins was demonstrated in Ambrosia, Betula, Cynodon, Helianthus, Mercurialis, Plantago and Phleum extracts by immunoblot inhibition studies. Purified Pho d 2 could be used for diagnosis and treatment of pollen profilin sensitization. RationaleProfilins are considered as an important panallergen being the IgE cross-reactivity among these proteins associated with multiple pollen sensitization. Sequence homology among pollen profilins is very high, ranging 67.4-89.5%. The objective was to evaluate the cross-reactivity among profilin from palm tree pollen (Phoenix dactylifera) and other pollen allergenic sources where profilin has been described. Profilins are considered as an important panallergen being the IgE cross-reactivity among these proteins associated with multiple pollen sensitization. Sequence homology among pollen profilins is very high, ranging 67.4-89.5%. The objective was to evaluate the cross-reactivity among profilin from palm tree pollen (Phoenix dactylifera) and other pollen allergenic sources where profilin has been described. MethodsPalm tree profilin (Pho d 2) was purified by affinity chromatography in a poly-L-proline column, sequenced by LC/MS-MS and used to produce rabbit polyclonal antibodies. Identification of profilin in eleven pollen extracts (Ambrosia artemisiifolia, Artemisia vulgaris, Betula alba, Chenopodium album, Cynodon dactylon, Helianthus annus, Mercurialis perennis, Olea europaea, Parietaria judaica, Phleum pratense and Plantago lanceolata) was carried out by IgG ELISA and immunoblot using the polyclonal antibodies. Cross-reactivity studies were performed by IgG immunoblot inhibition assays, using the purified profilin for inhibition. Palm tree profilin (Pho d 2) was purified by affinity chromatography in a poly-L-proline column, sequenced by LC/MS-MS and used to produce rabbit polyclonal antibodies. Identification of profilin in eleven pollen extracts (Ambrosia artemisiifolia, Artemisia vulgaris, Betula alba, Chenopodium album, Cynodon dactylon, Helianthus annus, Mercurialis perennis, Olea europaea, Parietaria judaica, Phleum pratense and Plantago lanceolata) was carried out by IgG ELISA and immunoblot using the polyclonal antibodies. Cross-reactivity studies were performed by IgG immunoblot inhibition assays, using the purified profilin for inhibition. ResultsHighly purified profilin (>95%) from palm tree pollen (Pho d 2) with a molecular weight of 14 kDa was obtained. Identity and purity of profilin was confirmed by LC/MS-MS. Presence of profilin was confirmed in all the extracts, except Chenopodium, by direct ELISA (O.D.>0.1). Identification of profilin by IgG immunoblot, and cross-reactivity with Pho d 2 by immunoblot-inhibition, was observed in Ambrosia, Betula, Cynodon, Helianthus, Mercurialis, Plantago and Phleum extracts. Highly purified profilin (>95%) from palm tree pollen (Pho d 2) with a molecular weight of 14 kDa was obtained. Identity and purity of profilin was confirmed by LC/MS-MS. Presence of profilin was confirmed in all the extracts, except Chenopodium, by direct ELISA (O.D.>0.1). Identification of profilin by IgG immunoblot, and cross-reactivity with Pho d 2 by immunoblot-inhibition, was observed in Ambrosia, Betula, Cynodon, Helianthus, Mercurialis, Plantago and Phleum extracts. ConclusionsCross-reactivity among Pho d 2 and other profilins was demonstrated in Ambrosia, Betula, Cynodon, Helianthus, Mercurialis, Plantago and Phleum extracts by immunoblot inhibition studies. Purified Pho d 2 could be used for diagnosis and treatment of pollen profilin sensitization. Cross-reactivity among Pho d 2 and other profilins was demonstrated in Ambrosia, Betula, Cynodon, Helianthus, Mercurialis, Plantago and Phleum extracts by immunoblot inhibition studies. Purified Pho d 2 could be used for diagnosis and treatment of pollen profilin sensitization.