LOCALIZATION OF THE 2 MAJOR ALLERGENS IN RYE-GRASS POLLEN USING SPECIFIC MONOCLONAL-ANTIBODIES AND QUANTITATIVE-ANALYSIS OF IMMUNOGOLD LABELING

The intracellular localization of the two major allergens, Lol p I and Lol p IX, in rye-grass anthers was examined using monoclonal antibodi es FMCA1 (specific for Lol p I) and FMCA7 (specific for Lol p IX) with immunocytochemical techniques and quantitative analysis. A newly deve loped anhydrous fixation technique in a mixture of glutaraldehyde, par aformaldehyde and 2,2-dimethoxypropane followed by embedding in LR Gol d resin resulted in both improved infiltration of pollen grains compar ed with existing techniques and the Localization of these water-solubl e antigens in their original sites compared with diffusion artefacts f ollowing aqueous methods. After anhydrous fixation, Lol p I was predom inantly located in the electron-opaque regions of the cytosol of the v egetative cell of the tricellular pollen grains (24 counts mu m(-2)), whereas Lol p IX was detected mainly within starch granules (16 counts mu m(-2)) For both Lol p I and Lol p IX, similar labelling was detect ed in the cells of the endothecium and middle layer (18 counts mu m(-2 )), but none was found in the tapetal cells or orbicules.