IMMUNOGOLD PROBES FOR LIGHT AND ELECTRON-MICROSCOPIC LOCALIZATION OF OLE-E-I IN SEVERAL OLEACEAE POLLENS

We investigated the immunolocalization of the olive major allergen Ole e I and Ole e I-like proteins in pollen from several Oleaceae species [olive (Olea europaea), ash (Fraxinus excelsior), privet (Ligustrum v ulgaris), lilac (Syringa vulgare), and forsythia (Forsythia suspensa)] . Crossreactions among different pollens were found in enzyme immunoas says. For immunolocalization with light microscopy we used the silver enhancement technique with three monoclonal antibodies (1D8, 10H1, and 16G2) that recognize three different epitopes of the allergen Ole e I . Our findings show that the silver enhancement technique is very usef ul when several antibodies are to be used for rapid screening of diffe rent materials. MAb 10H1 gave the most precise results and was selecte d for further immunolocalization studies with transmission electron mi croscopy. The epitope recognized by this MAb was localized exclusively in the endoplasmic reticulum in olive pollen. In lilac, privet, and a sh pollen, most of the reactivity was also seen in the endoplasmic ret iculum; however, the 10H1 epitope was not detected in forsythia pollen .