Abstract

The chief goal of this project is to understand the genetic mechanisms involved in antibody formation. Many investigations are focusing on the mechanisms of somatic diversification of antibody genes; this project will define the starting library, the inherited set, of immunoglobulin heavy chain variable region, Igh-V, genes of the BALB/c mouse. These studies will investigate the biological, genetic, and evolutionary factors which determine its composition and structure. The genetic analysis will be both Mendelian and molecular. The Mendelian analysis will proceed by analysis of genetic recombination in and around the Igh complex locus and by preservation of recombinant chromosomes as Igh Recombinant mouse strains. In these strains the expression of antibody markers (idiotypes and allotypes), neighboring lymphocyte alloantigen genes, and certain lymphocyte interactions will be characterized to define their regulation, roles and interactions in the immune response. The molecular analysis will proceed by recombinant DNA cloning of Vh genes and Southern blot analysis to determine the number of Vh genes and their relationships in homologous gene families. Analysis of the Vh libraries of disparate strains such as C57BL/6, DBA/2, A/He and NZB will characterize the nature and extent of polymorphism and delimit the biological requirements and allowed flexibility of these genes. The structure of the locus will be revealed by analysis of the recombined Vh arrays in the Igh Recombinant strains. Evolutionary changes and pathways will be identified by examination of the Vh libraries of a wide variety of mouse strains and substrains to characterize the processes of mutation, expansion, and contraction of Vh gene families. The utilization of each Vh gene and family to express antibodies of various specificities will be assessed by mRNA dot blot analysis of hybridomas and myelomas of known antigen binding specificity. The effects of polymorphic structural differences in V, D, and J sequences on idiotype expression will be investigated in Igh Recombinant strains.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI023548-05
Application #
3135839
Study Section
Mammalian Genetics Study Section (MGN)
Project Start
1985-09-01
Project End
1990-07-31
Budget Start
1989-01-01
Budget End
1990-07-31
Support Year
5
Fiscal Year
1989
Total Cost
Indirect Cost

  Institution

Name
Medical Biology Institute
Department
Type
DUNS #
City
La Jolla
State
CA
Country
United States
Zip Code
92037

  Publications

Degner, Stephanie C; Verma-Gaur, Jiyoti; Wong, Timothy P et al. (2011) CCCTC-binding factor (CTCF) and cohesin influence the genomic architecture of the Igh locus and antisense transcription in pro-B cells. Proc Natl Acad Sci U S A 108:9566-71
Jhunjhunwala, Suchit; van Zelm, Menno C; Peak, Mandy M et al. (2008) The 3D structure of the immunoglobulin heavy-chain locus: implications for long-range genomic interactions. Cell 133:265-79
Retter, Ida; Chevillard, Christophe; Scharfe, Maren et al. (2007) Sequence and characterization of the Ig heavy chain constant and partial variable region of the mouse strain 129S1. J Immunol 179:2419-27
Pawlitzky, Inka; Angeles, Christina V; Siegel, Andrea M et al. (2006) Identification of a candidate regulatory element within the 5' flanking region of the mouse Igh locus defined by pro-B cell-specific hypersensitivity associated with binding of PU.1, Pax5, and E2A. J Immunol 176:6839-51
Norio, Paolo; Kosiyatrakul, Settapong; Yang, Qiaoxin et al. (2005) Progressive activation of DNA replication initiation in large domains of the immunoglobulin heavy chain locus during B cell development. Mol Cell 20:575-87
Yang, Qiaoxin; Riblet, Roy; Schildkraut, Carl L (2005) Sites that direct nuclear compartmentalization are near the 5' end of the mouse immunoglobulin heavy-chain locus. Mol Cell Biol 25:6021-30
Jones Tiffany, Linda; Riblet, Roy; Stein, Kathryn E (2003) The Sr1 gene that controls diversity of the anti-inulin antibody response maps to mouse chromosome 14. Immunogenetics 55:80-6
Chevillard, Christophe; Ozaki, Jennifer; Herring, Christopher D et al. (2002) A three-megabase yeast artificial chromosome contig spanning the C57BL mouse Igh locus. J Immunol 168:5659-66
Zhou, Jie; Ermakova, Olga V; Riblet, Roy et al. (2002) Replication and subnuclear location dynamics of the immunoglobulin heavy-chain locus in B-lineage cells. Mol Cell Biol 22:4876-89
Zhou, Jie; Ashouian, Nasrin; Delepine, Marc et al. (2002) The origin of a developmentally regulated Igh replicon is located near the border of regulatory domains for Igh replication and expression. Proc Natl Acad Sci U S A 99:13693-8

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