Abstract

In this application, it is planned to identify molecular mechanisms of VH recombinatorial events involved in generation of the Ig heavy chains. Based on the application of FACS and PCR technology to the analysis of gene segment rearrangements in single cells isolated at various stages of B-cell development, the investigator will investigate the hypothesis that because H chains differ from clone to clone, B-cell development is clone specific. More to the point, while most of the molecular machinery responsible for B-cell differentiation is common to all B-cells, individual physicochemical characteristics of mu heavy chains can elicit positive or negative selective effects on the life of a B-cell. He proposes that a spectrum of heavy chains, even those which result from functional rearrangements, are dysfunctional and fail to mediate clonal maturation and allelic exclusion, particularly based on their ability to assemble with surrogate light chains. This hypothesis will be tested by transfection of a variety of genetically manipulated mu heavy chains into in vitro cell lines followed by analysis of their ability to assemble with surrogate light chains. Transgenic mice expressing so-called functional and dysfunctional heavy chains will be constructed to assess where these blocks in development occur.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI015797-20
Application #
2442398
Study Section
Special Emphasis Panel (ZRG2-IMS (01))
Project Start
1978-07-01
Project End
2001-06-30
Budget Start
1997-07-01
Budget End
1998-06-30
Support Year
20
Fiscal Year
1997
Total Cost
Indirect Cost

  Institution

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

  Publications

Diaz, Marilyn; Watson, Nicholas B; Turkington, Gene et al. (2003) Decreased frequency and highly aberrant spectrum of ultraviolet-induced mutations in the hprt gene of mouse fibroblasts expressing antisense RNA to DNA polymerase zeta. Mol Cancer Res 1:836-47
Diaz, M; Verkoczy, L K; Flajnik, M F et al. (2001) Decreased frequency of somatic hypermutation and impaired affinity maturation but intact germinal center formation in mice expressing antisense RNA to DNA polymerase zeta. J Immunol 167:327-35
Kline, G H; Hartwell, L; Beck-Engeser, G B et al. (1998) Pre-B cell receptor-mediated selection of pre-B cells synthesizing functional mu heavy chains. J Immunol 161:1608-18
Decker, D J; Linton, P J; Zaharevitz, S et al. (1995) Defining subsets of naive and memory B cells based on the ability of their progeny to somatically mutate in vitro. Immunity 2:195-203
Decker, D J; Kline, G H; Hayden, T A et al. (1995) Heavy chain V gene-specific elimination of B cells during the pre-B cell to B cell transition. J Immunol 154:4924-35
Stillman, C A; Linton, P J; Koutz, P J et al. (1994) Specific immunoglobulin cDNA clones produced from hybridoma cell lines and murine spleen fragment cultures by 3SR amplification. PCR Methods Appl 3:320-31
Linton, P J; Lo, D; Lai, L et al. (1992) Among naive precursor cell subpopulations only progenitors of memory B cells originate germinal centers. Eur J Immunol 22:1293-7
Linton, P J; Klinman, N R (1992) The generation of memory B cells. Semin Immunol 4:3-9
Decker, D J; Boyle, N E; Klinman, N R (1991) Predominance of nonproductive rearrangements of VH81X gene segments evidences a dependence of B cell clonal maturation on the structure of nascent H chains. J Immunol 147:1406-11
Decker, D J; Boyle, N E; Koziol, J A et al. (1991) The expression of the Ig H chain repertoire in developing bone marrow B lineage cells. J Immunol 146:350-61

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