The process by which T cells are activated, and the consequences of activation (e.g. new gene transcription, cytokine production, apoptosis), are being investigated by a variety of approaches: 1) activation-induced apoptosis of T cell hybridomas is mediated by upregulation of the ligand for Fas (FasL). We have identified a single major site 5' of the transcription initiation site that is required for initiation of fasL transcription. This site is bound by the Egr family of transcription factors, and Egr-2 and Egr-3 (but not Egr-1) are responsible for transactivation of gene transcription at this site. 2) We have found that corticosteroids prevent activation-induced apoptosis, and have hypothesized that this phenomenon regulates antigen-specific thymocyte selection. We have crossed transgenic mice that express antisense glucocorticoid receptor in immature thymocytes with lpr autoimmune mice, and have found that TCR Vbeta usage is altered and autoimmunity and lymphadenopathy are greatly diminished. Moreover, B10.BR (H-2k) mice bearing this antisense transgene no longer respond to the antigen pigeon cytochrome c, although they respond normally to more complex antigens such as PPD and alloantigen. These results demonstrate that glucocorticoids, likely those produced in the thymus itself, do in fact regulate positive and negative selection by antagonizing TCR-mediated signals, and therefore shape the peripheral immune repertoire. Studies on the possible role of glucocorticoids in the generation of autoimmunity are ongoing. 3) Inhibitors of proteasome activity block thymocyte apoptosis. We have found that IAPs (inhibitors of apoptosis) are selectively degraded in proteasomes in response to apoptotic stimuli. The IAPs themselves have ubiquitin protein ligase (E3) activity, and are responsible for their own ubiquitination. How this is regulated in vivo is under investigation. 4) Glucocorticoids are immunosuppressive, largely because they interfere with the transcription of many activation-induced genes. We are examining the possible role of a glucocorticoid-induced gene, GILZ, in this activity.

Agency
National Institute of Health (NIH)
Institute
Division of Basic Sciences - NCI (NCI)
Type
Intramural Research (Z01)
Project #
1Z01BC009290-15
Application #
6433155
Study Section
(LICB)
Project Start
Project End
Budget Start
Budget End
Support Year
15
Fiscal Year
2000
Total Cost
Indirect Cost
Name
Basic Sciences
Department
Type
DUNS #
City
State
Country
United States
Zip Code
Ashwell, Jonathan D (2006) The many paths to p38 mitogen-activated protein kinase activation in the immune system. Nat Rev Immunol 6:532-40
Schito, Marco L; Demidov, Oleg N; Saito, Shin'ichi et al. (2006) Wip1 phosphatase-deficient mice exhibit defective T cell maturation due to sustained p53 activation. J Immunol 176:4818-25
Wu, Chuan-Jin; Conze, Dietrich B; Li, Tao et al. (2006) Sensing of Lys 63-linked polyubiquitination by NEMO is a key event in NF-kappaB activation [corrected] Nat Cell Biol 8:398-406
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Wu, Chuan-Jin; Conze, Dietrich B; Li, Xiaoming et al. (2005) TNF-alpha induced c-IAP1/TRAF2 complex translocation to a Ubc6-containing compartment and TRAF2 ubiquitination. EMBO J 24:1886-98
Munitic, Ivana; Ryan, Philip E; Ashwell, Jonathan D (2005) T cells in G1 provide a memory-like response to secondary stimulation. J Immunol 174:4010-8
Salvador, Jesus M; Mittelstadt, Paul R; Belova, Galina I et al. (2005) The autoimmune suppressor Gadd45alpha inhibits the T cell alternative p38 activation pathway. Nat Immunol 6:396-402
Mittelstadt, Paul R; Ashwell, Jonathan D (2003) Disruption of glucocorticoid receptor exon 2 yields a ligand-responsive C-terminal fragment that regulates gene expression. Mol Endocrinol 17:1534-42
Yang, Y; Fang, S; Jensen, J P et al. (2000) Ubiquitin protein ligase activity of IAPs and their degradation in proteasomes in response to apoptotic stimuli. Science 288:874-7

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