Forty years of research in immunobiology have provided a sound yet rudimentary understanding of the most fundamental of all events in acquired immunity: clonal selection. Our proposed studies will focus on the cardinal elements that direct clonal development of B cell memory: somatic mutagenesis of antibody V genes, cell proliferation and selection. We will test the hypothesis 1) that somatic mutagenesis of antibody genes and selection for affinity improvements both occur within a specific subset of germinal center B cells that bear surface immunoglobulin, and 2) that positively selected cells enter a proliferation phase in which mutation ceases and immunoglobulin is not expressed. A major goal of our work is to isolate the mutating B cell. Our studies will be performed on B cells isolated ex vivo that are participating in an exceptionally well-defined immune response to the hapten p-azophenylarsonate, and that express a single combination of antibody variable (v) gene segments (VH, D, JH, VK, JH). A strength of the model is that these cells belong to lineages destined for the memory compartment. The feasibility of this proposal derives from recent advances in our laboratory that permit the isolation of single memory lineage cells from immune spleen, the direct sequence analysis and expression of their H and L V genes and V gene transcripts, and the production of single cell cDNA libraries. Germinal centers are sites for memory B cell development for the genesis of several common B cell neoplasias and for recruitment of autoreactive B cells. Results of these studies will advance our limited knowledge of regulatory processes that control mutation and proliferation in B cells by providing basic information at an unprecedented level of resolution. They will also position us to elucidate the mechanism of the only recognized physiologic mutator in the Animal Kingdom.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI039563-02
Application #
2390445
Study Section
Immunobiology Study Section (IMB)
Project Start
1996-04-01
Project End
2001-03-31
Budget Start
1997-04-01
Budget End
1998-03-31
Support Year
2
Fiscal Year
1997
Total Cost
Indirect Cost
Name
National Jewish Health
Department
Type
DUNS #
City
Denver
State
CO
Country
United States
Zip Code
80206
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Shapiro, Gary S; Ellison, Misoo C; Wysocki, Lawrence J (2003) Sequence-specific targeting of two bases on both DNA strands by the somatic hypermutation mechanism. Mol Immunol 40:287-95
Shapiro, Gary S; Aviszus, Katja; Murphy, James et al. (2002) Evolution of Ig DNA sequence to target specific base positions within codons for somatic hypermutation. J Immunol 168:2302-6
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Tumas-Brundage, K M; Notidis, E; Heltemes, L et al. (2001) Predominance of a novel splenic B cell population in mice expressing a transgene that encodes multireactive antibodies: support for additional heterogeneity of the B cell compartment. Int Immunol 13:475-84
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Shapiro, G S; Aviszus, K; Ikle, D et al. (1999) Predicting regional mutability in antibody V genes based solely on di- and trinucleotide sequence composition. J Immunol 163:259-68
Wysocki, L J; Liu, A H; Jena, P K (1998) Somatic mutagenesis and evolution of memory B cells. Curr Top Microbiol Immunol 229:105-31
Wysocki, L J; Zhang, X; Smith, D S et al. (1998) Somatic origin of T-cell epitopes within antibody variable regions: significance to monoclonal therapy and genesis of systemic autoimmune disease. Immunol Rev 162:233-46
Bonorino, C; Nardi, N B; Zhang, X et al. (1998) Characteristics of the strong antibody response to mycobacterial Hsp70: a primary, T cell-dependent IgG response with no evidence of natural priming or gamma delta T cell involvement. J Immunol 161:5210-6

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