Tob is a negative regulator of activation that is expressed in anergic and quiescent T cells

Abstract

During a search for genes that maintain T cell quiescence, we determined that Tob, a member of an anti-proliferative gene family, was highly expressed in anergic T cell clones. Tob was also expressed in unstimulated peripheral blood T lymphocytes and down-regulated during activation. Forced expression of Tob inhibited T cell proliferation and transcription of cytokines and cyclins. In contrast, suppression of Tob with an antisense oligonucleotide augmented CD3-mediated responses and abrogated the requirement of costimulation for maximal proliferation and cytokine secretion. Tob associated with Smad2 and Smad4 and enhanced Smad DNA-binding. The inhibitory effect of Tob on interleukin 2 (IL-2) transcription was not mediated by blockade of NFAT, AP-1 or NF-κB transactivation but by enhancement of Smad binding on the −105 negative regulatory element of the IL-2 promoter. Thus, T cell quiescence is an actively maintained phenotype that must be suppressed for T cell activation to occur.

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Figure 1: Northern blot analysis of Tob.
Figure 2: Tob inhibited T cell proliferation, cytokine transcription and cell cycle progression.
Figure 3: Elimination of Tob reduced the threshold of T cell activation.
Figure 4: Tob was expressed in the nucleus and associated with Smad4 and Smad2.
Figure 5: Tob enhanced Smad4 DNA-binding and transactivation.
Figure 6: Tob inhibited TCR + CD28–mediated IL-2 transcription, but not expression and transactivation of known IL-2 transcription factors.
Figure 7: Tob augmented Smad-mediated inhibition of IL-2 transcription by enhancing Smad binding on the IL-2 promoter.

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Acknowledgements

We thank K. Miyazono for the Smad2, Smad3 and Smad4 plasmids; W. Kruijer for the 4×SBE-luciferase plasmid; P. Coffer for the p27kip1 promoter-luciferase plasmid; P. van der Saag for the 4×NF-kB(HIV-LTR)tk-luciferase plasmid; and M. Greenberg for the pEF-LacZ plasmid. Supported by NIH grants AI 43552, AI 41584 and HL 54785.

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Correspondence to Vassiliki A. Boussiotis.

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Tzachanis, D., Freeman, G., Hirano, N. et al. Tob is a negative regulator of activation that is expressed in anergic and quiescent T cells. Nat Immunol 2, 1174–1182 (2001). https://doi.org/10.1038/ni730

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