Abstract

Cultured hybridoma cells producing monoclonal antibodies will be examined for their ability to generate mutant immunoglobulins with changes in their structure and function. Techniques have been developed to identify such mutants using monoclonals that recognize determinants on the different constant region domains. The frequency of such constant region mutations will be examined in both IgG and IgM producing hybridomas and compared with the frequency of variable region mutations in the same cell lines. Mutant monoclonal antibodies with changes in their intravascular equilibration and decay, in binding to Fc receptors, and in the fixation of complement will be identified and the structural bases of the mutations determined by DNA sequencing. The impact of such somatic mutations in the variable regions on H and L chain compatibility will also be studied. The effectiveness of representative mutant monoclonals in passive immunization and targeting to tumors will be evaluated. It is hoped that these experiments will not only provide insight into the instability of immunoglobulin genes and the structural basis of antibody function, but will also allow us to tailor-make monoclonal antibodies so that they will be more effective diagnostic and therapeutic reagents.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI005231-23
Application #
3124226
Study Section
Allergy and Immunology Study Section (ALY)
Project Start
1978-09-01
Project End
1988-08-31
Budget Start
1986-09-01
Budget End
1987-08-31
Support Year
23
Fiscal Year
1986
Total Cost
Indirect Cost

  Institution

Name
Albert Einstein College of Medicine
Department
Type
Schools of Medicine
DUNS #
009095365
City
Bronx
State
NY
Country
United States
Zip Code
10461

  Publications

Chien, N C; Roberts, V A; Giusti, A M et al. (1989) Significant structural and functional change of an antigen-binding site by a distant amino acid substitution: proposal of a structural mechanism. Proc Natl Acad Sci U S A 86:5532-6
Behar, S M; Fischer, B; Diamond, B et al. (1988) Variable region gene utilization and environmental antigens in autoimmunity. Monogr Allergy 24:297-306
Spira, G; Aguila, H L; Scharff, M D (1988) T15 PC-binding monoclonal antibodies retain specificity when they switch from IgM to IgG. J Immunol 140:2675-80
Pollock, R R; French, D L; Gefter, M L et al. (1988) Identification of mutant monoclonal antibodies with increased antigen binding. Proc Natl Acad Sci U S A 85:2298-302
Chien, N C; Pollock, R R; Desaymard, C et al. (1988) Point mutations cause the somatic diversification of IgM and IgG2a antiphosphorylcholine antibodies. J Exp Med 167:954-73
Harrington, D S; Peterson, C; Ness, M et al. (1988) Acute myelogenous leukemia with eosinophilic differentiation and trisomy-1. Am J Clin Pathol 90:464-9
French, D; Kelly, T; Buhl, S et al. (1987) Somatic cell genetic analysis of myelomas and hybridomas. Methods Enzymol 151:50-66
Giusti, A M; Chien, N C; Zack, D J et al. (1987) Somatic diversification of S107 from an antiphosphocholine to an anti-DNA autoantibody is due to a single base change in its heavy chain variable region. Proc Natl Acad Sci U S A 84:2926-30
Scharff, M D; Aguila, H L; Behar, S M et al. (1987) Studies on the somatic instability of immunoglobulin genes in vivo and in cultured cells. Immunol Rev 96:75-90
Scharff, M D; DePinho, R A; Behar, S et al. (1986) The role of monoclonal antibodies and the recombinant DNA technology in studying autoantibody production. Cell Immunol 99:29-37

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