Evaluation of allergenicity of the extract of Japanese juniper pollen reacting to sera from asthmatic children--by the methods of enzyme-linked immunosorbent assay and immunoblotting

We investigated the allergenicity of pollen extract of Japanese juniper (Juniperus rigida, Cupressaceae family) to sera of 49 asthmatic children by the methods of enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Three bands stained with Coomassie Brilliant Blue R were detected on SDS-PAGE. Sera from 27 (55.1%) out of the 49 children showed positive reaction to the pollen extract in ELISA. The same sera from the 27 children were used as the first antibody in immunoblotting, which confirmed the presence of a band of protein with 70 K dalton molecular weight (M.W) common to the all sera. This band was bound with concanavalin A in lectins. We successfully purified the antigenic substance of Japanese juniper pollen from this band by the electroelution method. The major allergen of Japanese juniper pollen is glycoprotein with 70 KM.W. On sandwich-ELISA, there was no reaction of Sugi Basic Protein (SBP) and anti-SBP to Japanese juniper is an allergen that has no cross-reactivity with SBP.