This is a second submission of a renewal project to study B-cell differentiation in immunoglobulin-transgenic scid mice. In preliminary experiments recently published, the investigator has found that in scid mice transgenic for Ig heavy chain genes, B lineage cells reach the pre-B-cell stage of development and that RAG1/2 levels are downregulated at the pro-B-cell stage. In Ig heavy and light chain gene transgenics, scid B lineage cells can reach the B-cell stage of development and RAG1/2 levels are down-regulated both at the pro-B and the pre-B-cell stages. Data from scid/+ and scid transgenics suggest that the heavy chain gene, alone, can prevent V-DJ joining events (but not DJ joining nor V-Jkappa joining events), while heavy and light chain genes prevent V-DJ joining and recombination at the kappa light chain locus, as well. The goals of the present application are to: 1) determine how an assembled heavy chain gene down-regulates RAG expression (and V-DJ joining) in pro-B-cells. It will be determined whether pseudo-light chain plays a role in this signaling and attempts will be made to clone genes involved in the signaling. 2) Determine why some mu/kappa gene combinations work to promote scid B-cell development while others do not. An hypothesis to be tested is whether this is a function of the mu,kappa combination's being able to arrest DNA recombination events which lead to unresolved breaks in scid cell DNA. 3) Explore the notion that the antigen-specificity of the mu,kappa combination determines whether it will down-regulate DNA recombination (and RAG expression). 4) Determine whether B-cell development is accelerated in Ig transgenic mice. 5) Assess the functional activity of scid B-cells in transgenic animals. Notably, these animals lack T-cells, like RAG-/- mice, but they remain """"""""leaky,"""""""" allowing low-frequency Ig gene rearrangements to take place. Questions will be asked about the nature of Ig-secreting cells that arise in these mice (e.g., express kappa transgene or endogenous kappa genes?) and whether B-cells with anti-self Ig transgenes (dsDNA-specificity) are able to develop in these animals. 6) Differences among mu,kappa transgenic scid mice have already been observed: the B-cells in some are responsive to mitogen while the B-cells in others are not. Attempts will be made to clone the genes that, when differentially expressed, allow for mitogen-responsiveness or mitogen-unresponsiveness, respectively.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA004946-35
Application #
2390592
Study Section
Immunobiology Study Section (IMB)
Project Start
1983-12-01
Project End
2000-03-31
Budget Start
1997-04-01
Budget End
1998-03-31
Support Year
35
Fiscal Year
1997
Total Cost
Indirect Cost
Name
Institute for Cancer Research
Department
Type
DUNS #
872612445
City
Philadelphia
State
PA
Country
United States
Zip Code
19111
Nakajima, Pamela B; Kiefer, Kerstin; Price, Amy et al. (2009) Two distinct populations of H chain-edited B cells show differential surrogate L chain dependence. J Immunol 182:3583-96
Kiefer, Kerstin; Nakajima, Pamela B; Oshinsky, Jennifer et al. (2008) Antigen receptor editing in anti-DNA transitional B cells deficient for surface IgM. J Immunol 180:6094-106
Bosma, Gayle C; Oshinsky, Jennifer; Kiefer, Kerstin et al. (2006) Development of functional B cells in a line of SCID mice with transgenes coding for anti-double-stranded DNA antibody. J Immunol 176:889-98
Bosma, Gayle C; Kim, Jiyoon; Urich, Teresa et al. (2002) DNA-dependent protein kinase activity is not required for immunoglobulin class switching. J Exp Med 196:1483-95
Nakajima, Pamela B; Bosma, Melvin J (2002) Variable diversity joining recombination: nonhairpin coding ends in thymocytes of SCID and wild-type mice. J Immunol 169:3094-104
Ruetsch, N R; Bosma, G C; Bosma, M J (2000) Unexpected rearrangement and expression of the immunoglobulin lambda1 locus in scid mice. J Exp Med 191:1933-43
Bosma, G C; Chang, Y; Karasuyama, H et al. (1999) Differential effect of an Ig mu transgene on development of pre-B cells in fetal and adult SCID mice. Proc Natl Acad Sci U S A 96:11952-7
Wiest, D L; Berger, M A; Carleton, M (1999) Control of early thymocyte development by the pre-T cell receptor complex: A receptor without a ligand? Semin Immunol 11:251-62
Chang, Y; Bosma, M J; Bosma, G C (1999) Extended duration of DH-JH rearrangement in immunoglobulin heavy chain transgenic mice: implications for regulation of allelic exclusion. J Exp Med 189:1295-305
Chang, Y; Bosma, M J (1997) Effect of different Ig transgenes on B cell differentiation in scid mice. Int Immunol 9:373-80

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