Major projects in this laboratory are all designed to further our understanding of allergens and to approach questions of fundamental interest in immunology using well defined allergens as a particularly relevant model. Using selected ragweed and grass pollen allergens isolated in this laboratory and elsewhere, the proteins are being characterized structurally (amino acid sequence analysis) and antigenically (by defining epitopes reactive with human IgE). The allergens currently under study include short ragweed Ra5, Ra6, and antigen E. From a series of grass pollens, the Group I allergen is the model system chosen for development. In addition to these molecules which are isolated from pollen in this laboratory, Western ragweed Ra5 (isolated elsewhere) will be sequenced in order to compare it with short and giant ragweed Ra5's whose structures have already been established. Monoclonal antibodies continue to be generated against allergens. Many of these have potential use in standardizing allergen extracts and many are useful in immunoassays for identifying epitopes recognized by human IgE and IgG antibodies. Currently, the monoclonal antibodies in use in the laboratory are of murine origin, but recent progress in using the Epstein Barr virus to """"""""immortalize"""""""" human antibody producing lymphocytes is encouraging and will be pursued. Acquisition of peptide synthesis capability adds a new dimension to studies of allergen epitopes. It is now possible to synthesize particular peptides, immunize experimental animals with these, and ascertain whether the immune serum reacts with the intact allergen. Finally, the ability to probe for and isolate the genes which code for allergens will enable this laboratory to begin to use powerful molecular biology approaches in the study of allergy and allergens. Enough is known about the structure of a number of allergens to allow the design of oligonucleotide probes which will anneal to genomic and cDNA libraries under construction. The family of monoclonal antibodies to these allergen allow isolation of polysomes actively synthesizing particular allergens in in vitro translation systems. """"""""Western"""""""" and """"""""dot"""""""" blotting procedures ensure that a particular message is present prior to synthesizing a cDNA library for sequence analysis and placement into expression vectors.
