The goal of these studies is an in-depth investigation of the thymus derived lymphocyte and its products. Lymphotoxin (LT), a product of antigen or mitogen activated T cells, is being studied as a model lymphokine. The long term goals are to study LT's cell of origin, biochemical nature, mechanism of action, relationship to other lymphokines, regulation and role in vivo. Murine lymphotoxin cDNA has been cloned with mRNA from an interleukin-2 maintained T cell clone. Its gene has been isolated and mapped to chromosome 17.
The specific aims of the next 5 years are to analyze the structure of the LT gene, to precisely localize it on chromosome 17, to determine the linkage relationship of the genes for LT and tumor necrosis factor (TNF), to obtain maximal expression of LT cDNA, to analyze regulation of LT and TNF, to study expression of the transfected LT gene and to analyze the 5' regulatory sequence of the LT gene. Goals will be accomplished by sequencing the gene, by identifying LT promoter activity, and by expressing LT DNA in prokaryotic and eurkaryotic cells. Interleukin-2 maintained T cell clones will be exposed to several inducing agents and their LT and TNF mRNA analyzed by Northern blots. The murine LT gene will be ligated to strong promoters and its expression studied in human T cell tumors. Similar vectors will be used to study the effect of LT gene overproduction in transgenic mice. Such mice will be analyzed for autoimmune pathology. If the LT gene is overexpressed in the T cells of these mice, and those cells killed, a murine model for AIDS will be available. LT 5' regulatory sequences will be ligated to a neutral gene, chloramphenical acetyle transferase, in order to study regulation of the LT gene in the absence of effects attributable to the product itself. Studies with transfected T cells, B cells, macrophages and transgenic mice will provide insigt into the nature of T cell specificity of LT expression and eventually lead to an understanding of the molecular basis of LT gene activation by antigen.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA016885-14
Application #
3164556
Study Section
Allergy and Immunology Study Section (ALY)
Project Start
1978-01-01
Project End
1992-06-30
Budget Start
1989-07-01
Budget End
1990-06-30
Support Year
14
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Yale University
Department
Type
Schools of Medicine
DUNS #
082359691
City
New Haven
State
CT
Country
United States
Zip Code
06520
Bentley, Kevin L; Stranford, Sharon; Liao, Shan et al. (2011) High endothelial venule reporter mice to probe regulation of lymph node vasculature. Adv Exp Med Biol 691:35-44
Akirav, Eitan M; Xu, Yan; Ruddle, Nancy H (2011) Resident B cells regulate thymic expression of myelin oligodendrocyte glycoprotein. J Neuroimmunol 235:33-9
Mounzer, Rawad H; Svendsen, Oyvind S; Baluk, Peter et al. (2010) Lymphotoxin-alpha contributes to lymphangiogenesis. Blood 116:2173-82
Akirav, Eitan M; Bergman, Cheryl M; Hill, Myriam et al. (2009) Depletion of CD4(+)CD25(+) T cells exacerbates experimental autoimmune encephalomyelitis induced by mouse, but not rat, antigens. J Neurosci Res 87:3511-9
Nasr, I W; Reel, M; Oberbarnscheidt, M H et al. (2007) Tertiary lymphoid tissues generate effector and memory T cells that lead to allograft rejection. Am J Transplant 7:1071-9
Mounzer, Rawad; Shkarin, Pavel; Papademetris, Xenophon et al. (2007) Dynamic imaging of lymphatic vessels and lymph nodes using a bimodal nanoparticulate contrast agent. Lymphat Res Biol 5:151-8
Liao, Shan; Bentley, Kevin; Lebrun, Marielle et al. (2007) Transgenic LacZ under control of Hec-6st regulatory sequences recapitulates endogenous gene expression on high endothelial venules. Proc Natl Acad Sci U S A 104:4577-82
Liao, Shan; Ruddle, Nancy H (2006) Synchrony of high endothelial venules and lymphatic vessels revealed by immunization. J Immunol 177:3369-79
Drayton, Danielle L; Liao, Shan; Mounzer, Rawad H et al. (2006) Lymphoid organ development: from ontogeny to neogenesis. Nat Immunol 7:344-53
Baddoura, Fady K; Nasr, Isam W; Wrobel, Barbara et al. (2005) Lymphoid neogenesis in murine cardiac allografts undergoing chronic rejection. Am J Transplant 5:510-6

Showing the most recent 10 out of 51 publications