Experiments are planned for a detailed analysis of the regulation of the rearrangement of immunoglobulin genes. Previous work with transgenic mice has shown that after rearrangement of a functional heavy and light chain gene a feedback mechanism shuts off the further production of the Ig gene specific recombinase. 1) It will now be determined whether the same mechanism operates in all types of B cells, or whether two different B cell lineages exist with respect to control mechanisms at the pre-B cell stage. 2) The role of gamma 2b in feedback inhibition has been controversial, apparently because the timing and/or quantities and/or membrane versus secreted forms of transgenic gamma 2b varied in different transgenic lineages. The different transgenic gamma 2b lines will now be exploited to shed more light on these parameters of the feedback control. 3) The isotypic exclusion of kappa and lambda genes will be further studied by determining the location of the lambda gene enhancer(s) and then producing transgenic mice with a gene under control of its own enhancer. 4) In order to define the molecular composition of the signal for feedback inhibition, recombinase positive pre-cell lines will be transfected with modified immunoglobulin genes and their effect on the recombinase will be determined. 5) Individual pre-cells from Ig gene transgenic mice will be analyzed for the timing of the expression of transgenic and endogenous Ig genes using the polymerase chain reaction. This may help unravel the kinetics of the feedback response. 6) By targeting the lambda 5 or C mu gene in embryonic stem cells with mutated gene constructs, mice will be produced which lack lambda 5 or mu production in order to study the role of these genes in early B cell development. It is hoped that these experiments may help in defining the control of immunoglobulin gene rearrangement and thus solve a well-known example of a developmental control, as well as be of clinical importance for immunological disorders with a B cell component.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD023089-06
Application #
3323090
Study Section
Immunobiology Study Section (IMB)
Project Start
1990-04-01
Project End
1995-03-31
Budget Start
1991-04-01
Budget End
1992-03-31
Support Year
6
Fiscal Year
1991
Total Cost
Indirect Cost
Name
University of Chicago
Department
Type
Schools of Medicine
DUNS #
225410919
City
Chicago
State
IL
Country
United States
Zip Code
60637
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Kurtz, B S; Witte, P L; Storb, U (1997) Gamma 2b provides only some of the signals normally given via mu in B cell development. Int Immunol 9:415-26
Roth, P E; Kurtz, B; Lo, D et al. (1995) lambda 5, but not mu, is required for B cell maturation in a unique gamma 2b transgenic mouse line. J Exp Med 181:1059-70
Weng, A; Engler, P; Storb, U (1995) The bulk chromatin structure of a murine transgene does not vary with its transcriptional or DNA methylation status. Mol Cell Biol 15:572-9
Doglio, L; Kim, J Y; Bozek, G et al. (1994) Expression of lambda and kappa genes can occur in all B cells and is initiated around the same pre-B-cell developmental stage. Dev Immunol 4:13-26
Storb, U; Roth, P; Kurtz, B K (1994) Gamma 2b transgenic mice as a model for the role of immunoglobulins in B cell development. Immunol Res 13:291-8
Engler, P; Weng, A; Storb, U (1993) Influence of CpG methylation and target spacing on V(D)J recombination in a transgenic substrate. Mol Cell Biol 13:571-7
Storb, U (1993) Steps in the generation of autoantibodies. Ann N Y Acad Sci 681:29-32
Roth, P E; Doglio, L; Manz, J T et al. (1993) Immunoglobulin gamma 2b transgenes inhibit heavy chain gene rearrangement, but cannot promote B cell development. J Exp Med 178:2007-21
Storb, U; Engler, P; Klotz, E et al. (1992) Rearrangement and expression of immunoglobulin genes in transgenic mice. Curr Top Microbiol Immunol 182:137-41

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