Elicitation and Characterization of Antibodies With Idiotypic (Ab1) and Anti-Idiotypic (Ab2) Reactivity: Epibodies

117 p.Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 1997.Studies of the regulatory idiotypic network have routinely assayed idiotypic (Ab1) and anti-idiotypic (Ab2) levels in the immune response. This work is a follow up of a previous study in which Ab1 and Ab2 responses to the hapten DNP were found to be similar in terms of magnitude and kinetics following immunization. In this study, spleen cells of mice immunized with different dinitrophenylated carriers were fused with myeloma cells to generate monoclonal antibodies. These were assayed by ELISA for Ab1 and Ab2 reactivity. It was found that some of antibodies bound only the antigen, others bound both antigen and idiotypic antibody, while others bound both antigen and idiotypic antibody, in addition to a wide range of unrelated antigens commonly used to demonstrate polyreactivity. Which type of reactivity depended on the carrier used, as well as the intensity of immunization. The monoclonal antibodies which bound only the antigen and idiotypic antibody (epibodies) were further characterized for such properties as specificity of binding, inhibition by various inhibitors, self-binding, and affinity of binding. The epibodies generated were specific for the binding site of the idiotypic antibody used for coating ELISA plates, and were inhibited by both hapten and idiotypic antibody. They had a potential for self-binding and had moderately high affinity for both antigen and idiotypic antibody. In this study, epibodies were therefore elicited in a conventional immune response.U of I OnlyRestricted to the U of I community idenfinitely during batch ingest of legacy ETD