The major goal of this grant and its predecessor is to study normal B lymphocyte development in the context of activation and expression of lymphocyte-specific genes. In the first grant the major focus was the rearrangement of immunoglobulin heavy and light chain genes during normal B cell development. This study included the isolation and sequencing of the light chain VK24 variable region gene family to determine their evolutionary relationship. In this proposal we expand from a study of immunoglobulin gene expression in normal lymphocytes to include the analysis of other B cell expressed genes. Using the techniques lineage tracing the growth and differentiation of pre-B cells shall be defined. At each stage of antigen-independent and antigen-dependent B cell differentiation the molecular genetic events relating to the heavy chain class switch shall be monitored on the expressed and on the nonexpressed heavy chain chromosome. This will require an analysis of these heavy chain arrangements in 1) three different stages of pre-B cell development; 2) in virgin BMu and mature B MuDelta cells; 3) in antigen-activated germinal center B cells expressing IgG or IgA receptor immunoglobulins of either one parental allotype or the other; and 4) in plasma cells. The lymphocyte differentiation antigens which are expressed in pre-B cell development (TdT, B220), in subsets of resting mature B cells (Ly-1, B220), and in antigen activated germinal center cells (transferrin receptor) are the primary objects of a new cDNA cloning strategy developed in this laboratory. The eventual objective of this cloning strategy is to characterize each of these genes in order to examine at the DNA sequence level whether particular shared sequences might be involved in their selective expression in B cells. Finally, a complete study of VK24 (antiphosphorycholine and antistreptococcyl A carbohydrate) light chain variable region genes will be carried out to understand their mechanism of evolution and their current status in mouse and man. The studies on normal pre-B cell and B cell differentiation should highlight those potential defects which may lead to B cell immunodeficiency syndromes. A study of the role of terminal transferase in providing immunoglobulin gene diversification during B cell differentiation will use as a model a mouse genetic defect (motheaten) whose hallmarks are immunodeficiency and autoimmunity. The studies on the immunoglobulin class switch mechanism should reveal important information for the eventual study of Ataxia Telangiectasia and selective immunoglobulin deficiency diseases. The analysis of the V kappa 24 family might reveal a basis for inordinate susceptibility to bacterial infection in certain humans.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI018162-08
Application #
3127732
Study Section
Immunobiology Study Section (IMB)
Project Start
1981-09-30
Project End
1989-11-30
Budget Start
1988-12-01
Budget End
1989-11-30
Support Year
8
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Stanford University
Department
Type
Schools of Medicine
DUNS #
800771545
City
Stanford
State
CA
Country
United States
Zip Code
94305
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Wu, Q; Tidmarsh, G F; Welch, P A et al. (1989) The early B lineage antigen BP-1 and the transformation-associated antigen 6C3 are on the same molecule. J Immunol 143:3303-8
Spangrude, G J; Klein, J; Heimfeld, S et al. (1989) Two monoclonal antibodies identify thymic-repopulating cells in mouse bone marrow. J Immunol 142:425-30
Okada, C Y; Weissman, I L (1989) Relative V beta transcript levels in thymus and peripheral lymphoid tissues from various mouse strains. Inverse correlation of I-E and Mls expression with relative abundance of several V beta transcripts in peripheral lymphoid tissues. J Exp Med 169:1703-19
Spangrude, G J; Muller-Sieburg, C E; Heimfeld, S et al. (1988) Two rare populations of mouse Thy-1lo bone marrow cells repopulate the thymus. J Exp Med 167:1671-83
Spangrude, G J; Weissman, I L (1988) Mature T cells generated from single thymic clones are phenotypically and functionally heterogeneous. J Immunol 141:1877-90
Spangrude, G J; Aihara, Y; Weissman, I L et al. (1988) The stem cell antigens Sca-1 and Sca-2 subdivide thymic and peripheral T lymphocytes into unique subsets. J Immunol 141:3697-707
Whitlock, C A; Tidmarsh, G F; Muller-Sieburg, C et al. (1987) Bone marrow stromal cell lines with lymphopoietic activity express high levels of a pre-B neoplasia-associated molecule. Cell 48:1009-21
Nottenburg, C; St John, T; Weissman, I L (1987) Unusual immunoglobulin DNA sequences from the nonexpressed chromosome of mouse normal B lymphocytes: implications for allelic exclusion and the DNA rearrangement process. J Immunol 139:1718-26
Muller-Sieburg, C E; Whitlock, C A; Weissman, I L (1986) Isolation of two early B lymphocyte progenitors from mouse marrow: a committed pre-pre-B cell and a clonogenic Thy-1-lo hematopoietic stem cell. Cell 44:653-62