Normal lymphocyte maturation is blocked in mutant C.B-17scid mice; consequently, they lack functional T and B cells. However, some scid mice are """"""""leaky"""""""" (approximately 15%); i.e., they spontaneously produced large quantities of immunoglobulin (Ig); preliminary evidence suggests that the B cells responsible are few in number and may all derive from a common B lymphocyte stem cell (B-LSC). We propose to test this possibility in several ways using Igh allotypes and novel Igh gene rearrangements as clonal markers. Assuming that monoclonality will apply to most leaky scid mice, we will evaluate the capacity of B-LSC for lymphocyte generation and replacement (persistence and quantity of Igh allotype levels) and diversity (number and kind of Igh-V genes expressed). This capacity will be compared to that of less committed hematopoietic stem cells (HSC). That is, we will deliberately make scid mice """"""""leaky"""""""" by reconstitution with B (and T) lymphocyte progency of a single normal HSC. Reconstitution will be accomplished by co-transfer of limiting numbers of BALB/c (Igh-a) and C.AL-20 (Igh-d) fetal liver cells into C.B-17scid (Igh-b) neonates (and embryos). Igh allotypes will serve as clonal markers. Varying the genotype and age of donors and recipients will enable us to examine both T and B cell lineages and the effect of different microenvironments on stem cell growth and differentiation. The value of the scid mouse model for the proposed study is that it enables us to assay the in vivo potential of stem cells at different stages of development and in the context of a normal developmental progression. Many reconstituted scid mice may show unrestricted lymphoid differentiation; i.e.; both T and B lymphocyte replacement. Leaky scid mice, on the other hand, should enable us to study restricted lymphoid differentiation; i.e., B lymphocyte generation in the absence of T cells and in the absence of recruitment of less committed HSC (functional T cells have not yet been detected in any scid mice).

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI013323-12
Application #
3125426
Study Section
Allergy and Immunology Study Section (ALY)
Project Start
1976-05-01
Project End
1991-05-31
Budget Start
1987-06-01
Budget End
1988-05-31
Support Year
12
Fiscal Year
1987
Total Cost
Indirect Cost
Name
Institute for Cancer Research
Department
Type
DUNS #
City
Philadelphia
State
PA
Country
United States
Zip Code
19111
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Chang, Y; Bosma, G C; Bosma, M J (1995) Development of B cells in scid mice with immunoglobulin transgenes: implications for the control of V(D)J recombination. Immunity 2:607-16
Fish, S M; Bosma, M J (1994) Abnormal deletions in the T-cell receptor delta locus of mouse thymocytes. Mol Cell Biol 14:4455-64
Kotloff, D B; Bosma, M J; Ruetsch, N R (1993) Scid mouse Pre-B cells with intracellular mu chains: analysis of recombinase activity and IgH gene rearrangements. Int Immunol 5:383-91
Kotloff, D B; Bosma, M J; Ruetsch, N R (1993) V(D)J recombination in peritoneal B cells of leaky scid mice. J Exp Med 178:1981-94
Roth, D B; Menetski, J P; Nakajima, P B et al. (1992) V(D)J recombination: broken DNA molecules with covalently sealed (hairpin) coding ends in scid mouse thymocytes. Cell 70:983-91
Roth, D B; Nakajima, P B; Menetski, J P et al. (1992) Double-strand breaks associated with V(D)J recombination at the TCR delta locus in murine thymocytes. Curr Top Microbiol Immunol 182:115-24
Bosma, M J (1992) B and T cell leakiness in the scid mouse mutant. Immunodefic Rev 3:261-76
Roth, D B; Nakajima, P B; Menetski, J P et al. (1992) V(D)J recombination in mouse thymocytes: double-strand breaks near T cell receptor delta rearrangement signals. Cell 69:41-53
Carroll, A M; Bosma, M J (1991) T-lymphocyte development in scid mice is arrested shortly after the initiation of T-cell receptor delta gene recombination. Genes Dev 5:1357-66

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