Immune tolerance is maintained, in part, by a process of clonal elimination in the population of immature lymphocyte. In particular, there is a well defined population of thymocytes that expresses both CD4 and CD8 (termed double positives or DPs), and this immature subset of thymocytes specifically undergoes apoptosis when challenged with antigen. The goal of this project is to define the biochemical mechanisms governing these events. The process of apoptosis is driven by the activation of caspases and the physiology of the mitochondria. The principal investigator plans to examine models of negative selection in the presence of endogenous and exogenously added inhibitors of caspases to determine which aspects of negative selection can be inhibited and whether forbidden self-reactive mature T cells appear in the thymus. The caspases present in the thymus and those that are activated as well as those absolutely required for antigen-mediated thymic deletion will be identified. Identified caspases will be the targets for genetic manipulation through the creation of deficient strains of mice. The impact of caspases on negative selection will be further studied by combining caspase deficiencies with the overexpression of Bcl-2, a protein that can act as a docking molecule and an ion channel regulating membrane potential of the mitochondria. Caspases will be further analyzed by transfection into a thymic cell line that mimics some of the aspects of thymocyte selection and development. Cell death in lymphocytes can be initiated by receptors in the TNFR family, and these receptors are known to act through the cytoplasmic mediator, FADD. Using mice that are deficient in FADD activity, preliminary data indicate that at least one model of negative selection is defective. Experiments are proposed to relate this defect to known cell surface and cytoplasmic mediators that bind FADD, and further experiments will identify FADD-interacting molecules that may be specifically expressed in the thymus. The long-term goals of this project are to establish a biochemical pathway that is important in the process of initiating apoptosis as a manifestation of clonal deletion in the thymus.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI037988-05
Application #
2887023
Study Section
Experimental Immunology Study Section (EI)
Program Officer
Quill, Helen R
Project Start
1995-06-01
Project End
2002-05-31
Budget Start
1999-06-01
Budget End
2000-05-31
Support Year
5
Fiscal Year
1999
Total Cost
Indirect Cost
Name
University of California San Diego
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
077758407
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Delpoux, Arnaud; Michelini, Rodrigo Hess; Verma, Shilpi et al. (2018) Continuous activity of Foxo1 is required to prevent anergy and maintain the memory state of CD8+ T cells. J Exp Med 215:575-594
Tsau, Jennifer S; Huang, Xin; Lai, Chen-Yen et al. (2018) The Effects of Dendritic Cell Hypersensitivity on Persistent Viral Infection. J Immunol 200:1335-1346
Lee, Sang-Yun; Coffey, Francis; Fahl, Shawn P et al. (2014) Noncanonical mode of ERK action controls alternative ?? and ?? T cell lineage fates. Immunity 41:934-46
Mingueneau, Michael; Krishnaswamy, Smita; Spitzer, Matthew H et al. (2014) Single-cell mass cytometry of TCR signaling: amplification of small initial differences results in low ERK activation in NOD mice. Proc Natl Acad Sci U S A 111:16466-71
Cuda, Carla M; Misharin, Alexander V; Gierut, Angelica K et al. (2014) Caspase-8 acts as a molecular rheostat to limit RIPK1- and MyD88-mediated dendritic cell activation. J Immunol 192:5548-60
Günther, Claudia; Martini, Eva; Wittkopf, Nadine et al. (2011) Caspase-8 regulates TNF-?-induced epithelial necroptosis and terminal ileitis. Nature 477:335-9
Ch'en, Irene L; Tsau, Jennifer S; Molkentin, Jeffery D et al. (2011) Mechanisms of necroptosis in T cells. J Exp Med 208:633-41
Hedrick, Stephen M; Ch'en, Irene L; Alves, Bryce N (2010) Intertwined pathways of programmed cell death in immunity. Immunol Rev 236:41-53
Ch'en, Irene L; Beisner, Daniel R; Degterev, Alexei et al. (2008) Antigen-mediated T cell expansion regulated by parallel pathways of death. Proc Natl Acad Sci U S A 105:17463-8
D'Souza, Warren N; Hedrick, Stephen M (2006) Cutting edge: latecomer CD8 T cells are imprinted with a unique differentiation program. J Immunol 177:777-81

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