Cellular caspase-8 (FLICE)-like inhibitory protein (c-FLIP) is an important regulator of death receptor-induced apoptosis and plays an essential role in thymocyte maturation. Two major isoforms of c-FLIP derived from alternative mRNA splicing, c-FLIPL and c-FLIPS, have been identified in mouse T lymphocytes. Our previous studies have demonstrated that conditional deletion of both c-FLIP isoforms in T lymphocytes results in an almost complete lack of mature T cells and increased apoptosis of single positive (SP) thymocytes. To further define the roles played by the c-FLIPL and c-FLIPS isoforms in thymocyte maturation and peripheral T cell function, we have generated mice specifically lacking the c-FLIPL (c-FLIPL-/-) or c-FLIPS (c-FLIPS-/-) isoform. Surprisingly, we found that expression of c-FLIPS but not c-FLIPL in c-FLIP conditional knockout mice rescued thymocyte development. Our studies further demonstrate that c-FLIPL is essential for mature T cell proliferation, as T cells from c-FLIPL-/- mice fail to develop into effectors after Listeria monocytogenes infection. Although accumulating evidence suggests that c-FLIP has both anti-apoptotic and cell signaling functions, the mechanisms by which c-FLIP regulates thymocyte maturation and mature T cell homeostasis remain unknown. Based on our preliminary results, we hypothesize that c-FLIP has three major functions mediated through c-FLIPL and c-FLIPS in the T cell compartment: 1. c-FLIPS protects mature SP thymocytes from TCR-induced apoptosis in the thymic medulla. 2. Both c-FLIP isoforms are essential in maintaining mature T cell homeostasis by promoting survival and proliferation. 3. c-FLIPL regulates T cell proliferation through its cleaved form c-FLIPp43. In this proposal, we will test these three hypotheses using several c-FLIP genetic models we have generated. The results will not only provide important insights into the mechanisms by which c-FLIP regulates thymocyte maturation and T cell homeostasis but also provide a better understanding of general T lymphocyte biology. Furthermore, determining the role of c-FLIP in regulating effector T cell survival may improve strategies for immunization and vaccine design. Narrative: c-FLIP is an important protein that protects T lymphocytes from death and is essential for T lymphocyte to develop. Our proposed studies will provide important information on how c-FLIP protects T cells and when it will protect T cells from death. Results from this study will improve our understanding of immunodeficiency and the regulation of immune response to microbial pathogen infections.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI073947-04
Application #
8014937
Study Section
Cellular and Molecular Immunology - B Study Section (CMIB)
Program Officer
Lapham, Cheryl K
Project Start
2008-03-01
Project End
2013-02-28
Budget Start
2011-03-01
Budget End
2012-02-29
Support Year
4
Fiscal Year
2011
Total Cost
$382,239
Indirect Cost
Name
Duke University
Department
Neurosciences
Type
Schools of Medicine
DUNS #
044387793
City
Durham
State
NC
Country
United States
Zip Code
27705
He, Ming-Xiao; He, You-Wen (2015) c-FLIP protects T lymphocytes from apoptosis in the intrinsic pathway. J Immunol 194:3444-51
Gordy, Claire; Liang, Jie; Pua, Heather et al. (2014) c-FLIP protects eosinophils from TNF-α-mediated cell death in vivo. PLoS One 9:e107724
He, Ming-Xiao; He, You-Wen (2013) CFLAR/c-FLIPL: a star in the autophagy, apoptosis and necroptosis alliance. Autophagy 9:791-3
He, M-X; He, Y-W (2013) A role for c-FLIP(L) in the regulation of apoptosis, autophagy, and necroptosis in T lymphocytes. Cell Death Differ 20:188-97
Piao, Xuehua; Komazawa-Sakon, Sachiko; Nishina, Takashi et al. (2012) c-FLIP maintains tissue homeostasis by preventing apoptosis and programmed necrosis. Sci Signal 5:ra93
Schattenberg, Jörn M; Wörns, Marcus A; Zimmermann, Tim et al. (2012) The role of death effector domain-containing proteins in acute oxidative cell injury in hepatocytes. Free Radic Biol Med 52:1911-7
He, Ming-Xiao; He, You-Wen (2012) Suppressing autoimmunity by TGF-β: not just through T(reg) cells. Cell Mol Immunol 9:371-2
McLeod, Ian X; He, You-Wen (2012) Editorial: TRPV1: how thymocytes sense stress and respond with autophagy. J Leukoc Biol 92:409-11
McLeod, Ian X; Jia, Wei; He, You-Wen (2012) The contribution of autophagy to lymphocyte survival and homeostasis. Immunol Rev 249:195-204
Paul, Suman; Kashyap, Anuj K; Jia, Wei et al. (2012) Selective autophagy of the adaptor protein Bcl10 modulates T cell receptor activation of NF-ýýB. Immunity 36:947-58

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