The hydrolysis of birch and timothy pollen allergen preparations by duodenal juice from an adult volunteer was studied in vitro with the aid of rocket immunoelectrophoresis, radiorocket immunoelectrophoresis and RAST-inhibition. The results achieved with all methods were similar. The duodenal juice had the capacity to hydrolyse both the grass and tree pollen proteins, and after 30 min preincubation only 10% of the original birch and timothy allergenic activity remained. Heat treatment of the duodenal juice completely destroyed the proteolytic activity (i.e. no hydrolysis occurred) while protease inhibitors only marginally prevented the hydrolysis.

Bee venom obtained by electrical stimulation has been analysed by crossed immunoelectrophoresis against high-titer rabbit antibodies. The antigenic analysis of bee venom revealed that the extract contained 17 antigens, which were detected in the crossed immunoelectrophoresis pattern after staining with Coomassie Brilliant Blue. The crossed immunoelectrophoresis pattern indicated heterogeneity in several of the antigens, as most of the precipitates (exhibiting enzyme and hemolytic activities) represented multiple forms of phospholipase A, hyaluronidase, acid phosphatase and melittin.

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Cultivated rye pollen extracts were characterized by means of. crossed immunoelectrophoresis (CIE) and crossed radioimmunoelectrophoresis (CRIE). 32 antigens were identified in CIE and among these 16 were radiostained in CRIE analysis of a rye pollen sensitive patient panel. Crossed line immunoelectrophoresis revealed that some of the rye pollen antigens were immunological partially identical with antigens of wheat flour and rye fluor.

Fourteen children with timothy grass pollinosis were given immunotherapy (IT) for 3 years with a purified and characterized timothy grass pollen preparation or a crude aqueous timothy pollen extract. Crossed radioimmunoelectrophoresis (CRIE) showed that 75% of the children under 11 years of age developed new specificities of IgE antibodies against timothy antigens, in contrast to older children, where no development of IgE antibodies against new timothy antigens could be detected. IgE antibodies were only detected against antigens formerly known as allergens. Timothy-specific IgG antibodies increased in most children during hyposensitization against the major allergens Ag 19 and Ag 24/25 and several other IgE-binding timothy antigens.