Demonstration of Birch Pollen Allergen from Isolated Pollen Grains Using Immunofluorescence and a Single Radial Immunodiffusion Technique

Abstract

Immunofiuorescence (IF) and single radial immunodiffusion (SRID) methods were modified in order to study the relation of allergen to the surface of birch pollen. By absorption of the anti-birch pollen sera employed, it was ascertained that both methods demonstrated thermostable allergenic birch pollen antigen and not nonallergenic antigens. The allergen rapidly diffuses from unfixed, wetted grains. After an aldehyde treatment and wet heating of the grains this leakage which mainly occurs through the pollen apertures was not demonstrated. Birch pollen allergen was released in a similar way from pollen of all of the six tested species of birch.

Localization of the major allergenBet vI in birch pollen by confocal laser scanning microscopy

“…After such a treatment excretion could probably only occur via the apertures. This theory is supported by the observations of Belin & Rowley (1971) of fluorescence around the apertures of birch pollen grains spread on agar gel. At present our theory is that when the dry birch pollen grains come into contact with the pistil they are equilibrated with the humidity on the stigmatic surface.…”

“…The reason for this assumption was that the allergens are released almost instantaneously when the grains come in contact with water. Belin & Rowley (1971) studied the liberation of allergenic proteins from birch pollen under different conditions. Before the treatment they fixed their samples on double adhesive tape, attached to microscope slides.…”

The role of the intine and cytoplasm in the activation and germination processes of Poaceae pollen grains

“…These proteins remained significantly in the intine and the labelling of exudates was scarce. Specific labelling in the activated Zwischenkörper of Lolium was also observed (Marquez et al 1997), just as allergens surrounding the pollen grains of Betula were observed by Belin & Rowley (1971). Recently Casas et al (1996) detected allergenic proteins in the intine and in material extruded from the activated pollen grains of Parietaria judaica, these proteins seem to be related to pollen-stigma recognition and they are activated after 10 min of hydration, prior to pollen tube formation.…”

Antibody Response in Rabbits Following Intranasal and Parenteral Administration of Birch Pollen

“…Immunodiffusion analysis of Sephadex G-100 fractions of birch pollen extract: a shows the analysis of unfractionated, crude extract (c) and the collected fractions 12-30, using polyspecific immune serum (placed in longitudinal basins). The antigens were analyzed before (upper rows) and after (lower rows) heating to 100 °C for 15 min, and b shows the analysis of pooled Sephadex G-100 fractions 11-17 (1), fractions 20-24 (2) and crude extract, heated to 100 C for 15 min (3) using polyspecific immune serum (4).…”