Studies are designed to determine the biochemical and genetic basis of diversity as it relates to affinity maturation in the immune response. In order to determine the basis of affinity monoclonal anti-fluorescyl antibodies of varying affinities and Ig class (i.e. IgG and IgM) will be characterized, grouped by idiotypic relatedness and primary structure studies initiated. Extensive studies into the properties, mechanisms and kinetics of ligand binding by each clone will be compiled for correlation with structural data (i.e. primary structure and crystallography). Kinetic studies will be expanded and refined to further the analyses of extended ligand dissociation rates which resolve into discrete groups with elapsed time suggesting programmed mutational events. Collectively all analyses will lead to understanding of the genetic mechanism (germ line vs. somatic mutation) of affinity maturation as a correlate to functional diversity found amongst antibody active sites. In vitro heavy and light chain recombination studies will be conducted with monoclonal antifluorescyl antibodies (IgG, IgM, Kappa and Lambda chains) of various affinities to determine the molecular basis for restricted association between chains. Experiments are designed to examine the extent of such restrictions and to understand cellular mechanisms responsible for translation and subsequent specific association of H and L chains. In vivo reconstitution studies will further elucidate the role of B and T (helper and suppressor) lymphocytes in the affinity maturation process. The understanding of mechanisms of affinity (binding) at the protein level and affinity maturation at the cellular level is important, if not critical, in development of improved vaccine development through eventual application of genetic engineering methodology. Selective control of low and high affinity may help to understand and control immune complex diseases prevalent in various disease processes such as autoimmunity. Affinity maturation is now important as a marker to explore clonal expansion and immunological defects.
Showing the most recent 10 out of 20 publications
