A major part of this project is directed toward the design and synthesis of highly specific and efficient immunomodulators. Effector molecules (e.g., monoclonal antibodies directed against designated cell surface proteins) are multiply and covalently linked to soluble polymer carriers of high molecular weight using heterobifunctional reagents. In some studies, protein-protein heteroconjugates have been prepared. New cross-linking reagents have been designed, synthesized and used as part of this project. Special emphasis has been given to improving the chemistry for functionalizing antibody molecules through their polysaccharide moieties and to the cross-linking of these derivatives to functionalized carriers. Collaborative studies employing the immunomodulators have dealt largely with a variety of cell surface factors involved in activation, growth and differentiation of B lymphocytes and presentation of antigen to T cells. For example, it has been shown (Finkelman, Mond et al.) that anti-IgD alone delivers a weak mitogenic signal to resting B cells, but when it is multiply linked to a soluble polymer such as dextran of MW > 200,000, strong mitogenic stimulation occurs at antibody concentrations ranging from 0.01 to 1.0 ng/ml. These conditions of stimulation are believed to closely approximate normal, T cell-independent antigenic signaling that appears to involve different pathways of transduction than believed to predominate for non-cross-linked signals at higher concentrations. Another objective of this project is to explore the property of general multispecificity of antibodies and other receptors. A study has been completed that measured the distribution of binding constants of a monoclonal antibody for a large collection of diverse compounds (not related to the homologous immunizing hapten). The results strongly support the statistical basis for multispecificity. Quantitative affinity chromatography was used to measure binding constants down to very small values. Techniques for preparing specialized affinity adsorbents were developed for these and other studies. Further work is planned for exploring alternative modes of complementation of ligands in receptor sites and relating these findings to specificity in immune systems and networks.
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| Golding, Basil; Eller, Nancy; Levy, Lily et al. (2002) Mucosal immunity in mice immunized with HIV-1 peptide conjugated to Brucella abortus. Vaccine 20:1445-50 |
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| Metzger, H; Chen, H; Goldstein, B et al. (1999) A quantitative approach to signal transduction. Immunol Lett 68:53-7 |
| Wofsy, C; Vonakis, B M; Metzger, H et al. (1999) One lyn molecule is sufficient to initiate phosphorylation of aggregated high-affinity IgE receptors. Proc Natl Acad Sci U S A 96:8615-20 |
