: We propose two aims in which Ig transgenic mice will be used to investigate the control of V gene expression in developing B-cells.
The first aim deals with the expression of VH genes, which differs dramatically in B-cells of adult and fetal mice. In developing B-cells of adult mice, the expression of VH genes is known to be dependent on the efficiency with which different H (mu) chains pair with L chains (surrogate or conventional) and on the antigen-binding specificity that results when a given mu and L chain pair. Whether VH expression is similarly dependent on such cellular selection in developing fetal B-cells is not known. To address this issue, we will selectively breed two lines of IgL chain transgenic mice (V-kappa-8 and V-lambda-l mice) lacking the surrogate light (SL) chain and endogenous L(kappa) chains. In this model, V-kappa-8 or V-lambda-l chains will substitute for the SL chain. Nascent u chains will essentially have the choice of only a single L chain (V-kappa-8 or V-lambda-l) with which to pair and form a B-cell receptor (BCR). We will test V-kappa-8 and V-lambda-l mice for usage of different VH genes, differential mu-L chain pairing and for evidence of B-cell selection based on BCR specificity. Using a novel assay system, we will also measure the relative frequency at which different VH gene families are targeted for rearrangement. In this way, we will assess whether the expression of different VH genes in fetal and adult B-cells primarily reflects different outcomes of cellular selection (hypothesis-1) or the possible absence of cellular selection in fetal B-cell development (hypothesis-2).
The second aim deals with the control of VL gene expression. We have observed a striking example of differential control of V-lambda-l/J-lambda-l rearrangement in two lines of IgH (mu) transgenic mice (M54 and 3H9 mice). In MM but not 3H9 mice, V-lambda-1/J-lambda-l rearrangement is suppressed. We hypothesize that the differential effect of these transgenes is due to a VH-related difference in the M54 and 3H9 mu chains or a tg-induced difference in gene expression between M54 and 3H9 B lineage cells. We will test these hypotheses using several different strategies. This will include testing whether the M54 mu chain, or any of the gene products identified by cDNA subtraction between M54 and 3H9 pre-B-cells, can inhibit temperature-induced V-lambda-1/J-lambda-1 rearrangement in transformed pm-B-cell lines.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
2R01CA004946-38A1
Application #
6328235
Study Section
Immunobiology Study Section (IMB)
Program Officer
Mccarthy, Susan A
Project Start
1983-12-01
Project End
2006-03-31
Budget Start
2001-04-01
Budget End
2002-03-31
Support Year
38
Fiscal Year
2001
Total Cost
$551,145
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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