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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Weiss, A. & Littman, D. R. Signal transduction by lymphocyte antigen receptors. Cell 76, 263–274 (1994).
Schwartz, R. H. T cell clonal anergy. Curr. Opin. Immunol. 9, 351–357 (1997).
Mueller, D. L., Jenkins, M. K. & Schwartz, R. H. An accessory cell-derived costimulatory signal acts independently of protein kinase C activation to allow T cell proliferation and prevent the induction of unresponsiveness. J. Immunol. 142, 2617–2628 (1989).
Boussiotis, V. A. et al. Prevention of T cell anergy by signaling through the γc chain of the IL-2 receptor. Science 266, 1039–1042 (1994).
Quill, H. et al. Anergic Th1 cells express altered levels of the protein tyrosine kinases p56lck and p59fyn. J. Immunol. 149, 2887–2893 (1992).
Gajewski, T. F., Qian, D., Fields, P. & Fitch, F. W. Anergic T-lymphocyte clones have altered inositol phosphate, calcium and tyrosine kinase signaling pathways. Proc. Natl Acad. Sci. USA 91, 38–42 (1994).
Sloan-Lancaster, J., Shaw, A. S., Rothbard, J. B. & Allen, P. M. Partial T cell signaling: Altered phospho-ζ and lack of Zap70 recruitment in APL-induced T cell anergy. Cell 79, 913–922 (1994).
Madrenas, J. et al. ζ phosphorylation without ZAP-70 activation induced by TCR antagonists or partial agonists. Science 267, 515–518 (1995).
Li, W., Whaley, C. D., Mondino, A. & Mueller, D. L. Blocked signal transduction to the ERK and JNK protein kinases in anergic CD4+ T cells. Science 271, 1272–1276 (1996).
Fields, P. E., Gajewski, T. F. & Fitch, F. W. Blocked Ras activation in anergic CD4+ T cells. Science 271, 1276–1278 (1996).
Boussiotis, V. A., Freeman, G. J., Berezovskaya, A., Barber, D. L. & Nadler, L. M. Maintenance of human T cell anergy: Blocking of IL-2 gene transcription by activated Rap1. Science 278, 124–128 (1997).
Boussiotis, V. A. et al. p27kip1 functions as an anergy factor inhibiting IL-2 transcription and clonal expansion of alloreactive human and murine helper T lymphocytes. Nature Med. 6, 290–297 (2000).
Greenwald, R. J., Boussiotis, V. A., Lorsbach, R. B., Abbas, A. K. & Sharpe, A. H. CTLA4 regulates peripheral T cell tolenance in vivo. Immunity 14, 145–155 (2001).
Diatchenko, L. et al. Suppression subtractive hybridization: A method for generating differentially regulated or tissue-specific cDNA probes and libraries. Proc. Natl Acad. Sci. USA 93, 6025–6030 (1996).
Bradbury, A., Possenti, R., Shooter, R. & Tirone, F. Molecular cloning of PC3, a putatively secreted protein whose mRNA is induced by nerve growth factor and depolarization. Proc. Natl Acad. Sci. USA 88, 3353–3357 (1991).
Fletcher, B. S. et al. Structure and expression of TIS21, a primary response gene induced by growth factors and tumor promoters. J. Biol. Chem. 266, 14511–14518 (1991).
Rimokh, R. et al. A chromosome 12 coding region is juxtoposed to myc protooncogene locus in a t(8;12)(q24;q22) translocation in a case of B-cell chronic lymphocytic leukemia. Genes Chrom. Cancer 3, 24–36 (1991).
Rouault, J.-P. et al. BTG1, a member of a new family of antiproliferative genes. EMBO J. 11, 1663–1670 (1992).
Varnum, B. C., Reddy, S. T., Koski, R. A. & Herschman, H. R. Synthesis, degradation and subcellular localization of protein encoded by the primary response genes TIS7/PC4 and TIS21/PC3. J. Cell. Physiol. 158, 205–213 (1994).
Rouault, J.-P. et al. Identification of BTG2, an antiproliferative p53-dependent component of the DNA damage cellular response pathway. Nature Genet. 14, 482–486 (1996).
Matsuda, S. et al. Tob, a novel protein that interacts with p185ebrB2, is associated with anti-proliferative activity. Oncogene 12, 705–713 (1996).
Yoshida, Y. et al. ANA, a novel member of Tob/BTG1 family, is expressed in the ventricular zone of the developing central nervous system. Oncogene 16, 2687–2693 (1998).
Ikematsu, N. et al. Tob2, a novel anti-proliferative Tob/BTG1 family member, associates with a component of the CCR4 transcriptional regulatory complex capable of binding cyclin-dependent kinases. Oncogene 18, 7432–7441 (1999).
Elliot, G. & O'Hare, P. Intercellular trafficing and protein delivery by a herpesvirus structural protein. Cell 88, 223–233 (1997).
DeCaprio, J. A., Furukawa, T., Ajchenbaum, F., Griffin, J. D. & Livingston, D. M. The retinoblastoma-susceptibility gene product becomes phosphorylated in multiple stages during cell cycle entry and progression. Proc. Natl Acad. Sci. USA 89, 1795–1798 (1992).
Medema, R. H., Kops, G. J., Bos, J. L. & Burgering, B. M. AFX-like Forkhead transcription factors mediate cell-cycle regulation by Ras and PKB through p27kip1. Nature 404, 782–787 (2000).
Viola, A., Schroeder, S., Sakakibara, Y. & Lanzavecchia, A. T lymphocyte costimulation mediated by reorganization of membrane microdomains. Science 283, 680–682 (1999).
Prevot, D. et al. Relationships of the antiproliferative proteins BTG1 and BTG2 with CAF1, the human homolog of a component of the yeast CCR4 transcriptional complex. J. Biol. Chem. 276, 9640–9648 (2001).
Draper, M. P. & Denis, C. L. Identification of a mouse protein whose homolog in Saccharomyces cerevisiae is a component of the CCR4 transcripitonal regulatory complex. Mol. Cell. Biol. 15, 3487–3495 (1995).
Yoshida, Y. et al. Negative regulation of BMP/Smad signaling by Tob in osteoblasts. Cell 103, 1085–1097 (2000).
Itoh, S., Itoh, F., Goumans, M.-J. & ten Dijke, P. Signaling of transforming growth factor-β family members through Smad proteins. Eur. J. Biochem. 267, 6954–6967 (2000).
Shull, M. M. et al. Targeted disruption of the mouse transforming growth factor-β 1 gene results in multifocal inflamatory disease. Nature 359, 693–699 (1992).
Gorelik, L. & Flavell, R. A. Abrogation of TGFβ signaling in T cells leads to spontaneous T cell differentiation and autoimmune disease. Immunity 12, 171–181 (2000).
Yang, X. et al. Targeted disruption of SMAD3 results in impaired mucosal immunity and diminished T cell responsiveness to TGF-β. EMBO J. 18, 1280–1291 (1999).
Nakao, A. et al. Blockade of transforming growth factor β/smad signaling in T cells by overexpression of Smad7 enhances antigen-induced airway inflammation and airway reactivity. J. Exp. Med. 192, 151–158 (2000).
Jonk, L. J. C., Itoh, S., Heldin, C.-H., ten Dijke, P. & Kruijer, W. Identification and functional characterization of a Smad binding element (SBE) in the JunB promoter that acts as a transforming growth factor-β, activin, and bone morphogenetic protein-inducible enhancer. J. Biol. Chem. 273, 21145–21152 (1998).
Freeman, G. J. et al. CTLA-4 and CD28 mRNAs are coexpressed in most activated T cells after activation: Expression of CTLA-4 and CD28 messenger RNA does not correlate with the pattern of lymphokine production. J. Immunol. 149, 3795–3801 (1992).
Agata, Y. et al. Expression of the PD-1 antigen on the surface of stimulated mouse T and B lymphocytes. Int. Immunol. 8, 765–772 (1996).
Nishimura, H., Nose, M., Hiai, H., Minato, N. & Honjo, T. Development of lupus-like autoimmune diseases by disruption of the PD-1 gene encoding an ITIM motif-carrying immunoreceptor. Immunity 11, 141–151 (1999).
Liu, H.-Y. et al. The Not proteins are part of CCR4 transcriptional complex and affect gene expression both positively and negatively. EMBO J. 4, 1096–1106 (1998).
Bucley, A. F., Kuo, C. T. & Leiden, J. M. Transcription factor LKLF is sufficient to program T cell quiescence via a c-Myc-dependent pathway. Nature Immunol. 2, 698–704 (2001).
Turner, J. & Crossley, M. Mammalian Kruppel-like transcription factors: more than just a pretty finger. Trends Biochem. Sci. 24, 236–241 (1999).
Kuo, C. T., Veselits, M. & Leiden, J. M. LKLF: A transcriptional regulator of single-positive T cell quiescence and survival. Science 277, 1986–1990 (1997).
Ghia, P. et al. Ordering of human bone marrow B lymphocyte precursors by single-cell polymerase chain reaction analysis of the rearrangement status of the immunoglobulin H and L chain gene loci. J. Exp. Med. 184, 2217–2229 (1996).
Schreiber, E., Matthias, P., Muller, M. M. & Schaffner, W. Rapid detection of octamer binding proteins with “mini-extracts”, prepared from a small number of cells. Nucleic Acid Res. 17, 6419–6420 (1989).
Nasevicius, A. & Ekker, S. Effective targeted gene “knockdown” in zebrafish. Nature Genet. 26, 216–220 (2000).
Dent, C. L. & Latchman, D. S. in Transcription Factors. A practical approach (ed. Latchman, D. S.)1–26 (Oxford University Press, NY, 1994).
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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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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