Terryn Hough

Cloning sequencing of Lol pI, the major allergenic protein of rye-grass pollen

We have isolated a full length cDNA clone encoding the major glycoprotein allergen Lol pI. The clone was selected using a combination of immunological screening of a cDNA expression library and PCR amplification of Lol pI‐specific transcripts. Lol pI expressed in bacteria as a fusion protein shows recognition by specific IgE antibodies present in sera of grass pollen‐allergic subjects. Northern analysis has shown that the Lol pI transcripts are expressed only in pollen of rye‐grass. Molecular cloning of Lol pI provides a molecular genetic approach to study the structure—function relationship of allergens.

Immunofluorescent screening of monoclonal antibodies to surface antigens of animal and plant cells bound to polycarbonate membranes

We have developed an immunofluorescent screening method in which animal or plant cells are immobilized without fixation on polycarbonate (Nuclepore) membranes. The technique does not involve centrifugation, and virtually no cells are lost during processing. Direct microscopic analysis of antibody binding provides an extremely sensitive and reproducible screening system for hybridoma supernatants.

Antibodies to pollen exines

A polyclonal antiserum and monoclonal antibodies have been prepared to purified pollen exines of Calocedrus decurrens Florin. The location of the antigen is in the exine, as shown by light-and electron-microscopic immunocytochemistry. The greatest reduction in antibody binding follows treatment of the exine with chemicals known to alter sporopollenin. These results provide evidence that sporopollenin is antigenic. Exines of ten species of gymnosperms and angiosperms also bound the polyclonal antiserum, indicating similarity of sporopollenin structure.

Pollen allergens: molecular cloning and mechanism for pollen-induced asthma.

Grass pollen is the major outdoor environmental factor provoking the immediate hypersensitive response of allergic disease in about one in five of the human population (see review by Howlett and Knox,1984). The symptoms are induced by the production of specific types of human defence molecules of the immune system, especially Immunoglubulin E (IgE). Allergens are environmental antigens which induce the formation of specific IgE, and bind specifically to IgE on the surface of epithelial mast cells. This triggers the mast cells to release the mediators of allergy.