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.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI020960-03
Application #
3130840
Study Section
Allergy and Immunology Study Section (ALY)
Project Start
1984-04-01
Project End
1988-03-31
Budget Start
1986-04-01
Budget End
1987-03-31
Support Year
3
Fiscal Year
1986
Total Cost
Indirect Cost
Name
University of Illinois Urbana-Champaign
Department
Type
Schools of Arts and Sciences
DUNS #
041544081
City
Champaign
State
IL
Country
United States
Zip Code
61820
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Voss Jr, E W; Dombrink-Kurtzman, M A; Ballard, D W (1989) Inter-relationship between immunoglobulin idiotype and metatype. Mol Immunol 26:971-7
Dombrink-Kurtzman, M A; Lacy, M J; Voss Jr, E W (1989) Structural properties of an anti-fluorescein monoclonal IgM cryoglobulin. Mol Immunol 26:779-87
Herron, J N; He, X M; Mason, M L et al. (1989) Three-dimensional structure of a fluorescein-Fab complex crystallized in 2-methyl-2,4-pentanediol. Proteins 5:271-80
Lacy, M J; Dombrink-Kurtzman, M A; Voss Jr, E W (1989) Quantitative analyses of immune network components. J Immunol 142:3482-8
Dombrink-Kurtzman, M A; Johnson, L S; Riordan, G S et al. (1989) Variable region primary structures of a high affinity anti-fluorescein immunoglobulin M cryoglobulin exhibiting oxazolone cross-reactivity. J Biol Chem 264:4513-22
Bedzyk, W D; Johnson, L S; Riordan, G S et al. (1989) Comparison of variable region primary structures within an anti-fluorescein idiotype family. J Biol Chem 264:1565-9
Lacy, M J; Voss Jr, E W (1989) Direct adsorption of ssDNA to polystyrene for characterization of the DNA/anti-DNA interaction, and immunoassay for anti-DNA autoantibody in New Zealand White mice. J Immunol Methods 116:87-98
Dombrink-Kurtzman, M A; Voss Jr, E W (1988) Cryoprecipitation properties of a high-affinity monoclonal IgM anti-fluorescyl antibody. Mol Immunol 25:1309-20
Voss Jr, E W; Dombrink-Kurtzman, M A; Miklasz, S D (1988) Functional and structural implications of variable region immunoglobulin dynamic states. Immunol Invest 17:25-39

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