The proposed experiments will address the problem of the ontogeny of T lymphocytes in adult and developing mice. Several new techniques based on retroviral-mediated gene transfer will be combined with the established methods of monoclonal antibody cell surface markers to examine the general questions of lineage relationships within the T cell compartment, tissue-specific expression of genes by T cell progenitors and immature T cells, and dynamics of T cell maturation during development. Replication-defective retroviruses will be used to infect murine bone marrow cells and these cells will be introduced into irradiated recipients. Each infection event by virtue of its unique position of chromosome insertion marks a stem cell and its progeny. Analysis of animals at various times after transplantation will allow estimation of numbers of progenitors involved in thymic and peripheral T cell repopulation. The distribution of clonal markers in monoclonal antibody-defined thymocyte subpopulations will also be studied. These experiments may allow the dissection of separate intrathymic lineages and inferences on ordered precursor-progeny relationships. Second, novel retrovirus vectors will be constructed with the goal of high-efficiency expression of transferred genes in thymocytes. Combination of this method with introduction of a transforming gene, SV40 large T-antigen, under the control of the adenosine deaminase (ADA) promoter may allow clonal expansion of the thymus subpopulation having high ADA expression. Third, by introduction of retroviruses into developing mouse fetuses, lymphocyte stem cell migration and cell fates will be explored. Use of a novel marker gene, E. coli lacZ, should allow in situ examination of marked thymocyte stem cells. Control of gene expression after retrovirus infection of postimplantation embryos will also be explored. These studies should allow definition of lineage relationships and stem cell regulation in the T cell compartment not heretofore possible. Better understanding of T cell development should have wide ranging implications for studies of immune function in genetic immune deficiencies, response to infectious agents, and in autoimmune disorders.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
1R29HD022880-01
Application #
3469483
Study Section
Immunological Sciences Study Section (IMS)
Project Start
1987-05-01
Project End
1992-04-30
Budget Start
1987-05-01
Budget End
1988-04-30
Support Year
1
Fiscal Year
1987
Total Cost
Indirect Cost
Name
Baylor College of Medicine
Department
Type
Schools of Medicine
DUNS #
074615394
City
Houston
State
TX
Country
United States
Zip Code
77030
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Allen, R C; Zoghbi, H Y; Moseley, A B et al. (1992) Methylation of HpaII and HhaI sites near the polymorphic CAG repeat in the human androgen-receptor gene correlates with X chromosome inactivation. Am J Hum Genet 51:1229-39
Fletcher, F A; Moore, K A; Ashkenazi, M et al. (1991) Leukemia inhibitory factor improves survival of retroviral vector-infected hematopoietic stem cells in vitro, allowing efficient long-term expression of vector-encoded human adenosine deaminase in vivo. J Exp Med 174:837-45
Aguilar, L K; Belmont, J W (1991) V gamma 3 T cell receptor rearrangement and expression in the adult thymus. J Immunol 146:1348-52
Moore, K A; Scarpa, M; Kooyer, S et al. (1991) Evaluation of lymphoid-specific enhancer addition or substitution in a basic retrovirus vector. Hum Gene Ther 2:307-15
Cournoyer, D; Scarpa, M; Mitani, K et al. (1991) Gene transfer of adenosine deaminase into primitive human hematopoietic progenitor cells. Hum Gene Ther 2:203-13
Fletcher, F A; Belmont, J W (1991) Stimulation of retroviral vector infection of murine hematopoietic progenitors. Int J Cell Cloning 9:491-502
Alford, R L; Belmont, J W (1990) Stable RNA secondary structure in a retroviral vector insert terminates reverse transcriptase elongation in vitro but not in cultured cells. Hum Gene Ther 1:269-76
Fletcher, F A; Williams, D E; Maliszewski, C et al. (1990) Murine leukemia inhibitory factor enhances retroviral-vector infection efficiency of hematopoietic progenitors. Blood 76:1098-103
Cournoyer, D; Scarpa, M; Jones, S N et al. (1990) Gene therapy: a new approach for the treatment of genetic disorders. Clin Pharmacol Ther 47:1-11

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