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T cell anergy is reversed by active Ras and is regulated by diacylglycerol kinase-α

Nature Immunology volume 7, pages 1166–1173 (2006)Cite this article

An Erratum to this article was published on 01 December 2006

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Abstract

T cell anergy has been correlated with defective signaling by the GTPase Ras, but causal and mechanistic data linking defective Ras activity with T cell anergy are lacking. Here we used adenoviral transduction to genetically manipulate nonproliferating T cells and show that active Ras restored interleukin 2 production and mitogen-activated protein kinase signaling in T cells that were made anergic in vitro or in vivo. Diacylglycerol kinases (DGKs), which negatively regulate Ras activity, were upregulated in anergic T cells, and a DGK inhibitor restored interleukin 2 production in anergic T cells. Both anergy and DGK-α overexpression were associated with defective translocation of the Ras guanine nucleotide–exchange factor RasGRP1 to the plasma membrane. Our data support a causal function for excess DGK activity and defective Ras signaling in T cell anergy.

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Figure 1: Constitutively active Ras restores IL-2 production in CAR TH1 cells made anergic in vitro.
Figure 2: Constitutively active Ras restores MAP kinase signaling and IL-2 promoter activity in TH1 cells made anergic in vitro.
Figure 3: Constitutively active Ras restores IL-2 production and MAP kinase activation in CAR CD8+ 2C T cells made anergic in vivo.
Figure 4: CrkL and Cbl function are not required for T cell anergy.
Figure 5: Expression of DGK-α is upregulated in anergic TH1 cells.
Figure 6: Defective localization of RasGRP1 to the plasma membrane in anergic 2C T cells.
Figure 7: Overexpression of DGK-α results in a state resembling anergy.
Figure 8: Overexpression of DGK-α results in diminished RasGRP1 plasma membrane translocation.
Figure 9: A DGK inhibitor partially restores IL-2 production by anergic TH1 cells.

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  • 03 November 2006

    In the version of this article initially published, the equal contribution of the third author was omitted. The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We thank J. DeGregori for the human ubiquitin C promoter; J. Washington for assistance with mouse breeding; and C. Kao, F. Rivas, R. Shah and S. Bond for experimental assistance. Supported the National Institutes of Health (R01AI7919, P01CA97296 and R21AI59818), the Arthritis Foundation (T.F.G.) and Deutsche Forschungsgemeinschaft (R.M.).

Author information

Author notes

  1. Reinhard Marks

    Present address: Department of Hematology and Oncology, University Hospital Freiberg, Freiberg, 79106, Germany

  2. Yuanyuan Zha, Reinhard Marks and Allen W Ho: These authors contributed equally to this work.

Authors and Affiliations

  1. Departments of Pathology and Medicine, Section of Hematology and Oncology, University of Chicago, Chicago, 60637, Illinois, USA

    Yuanyuan Zha, Reinhard Marks, Allen W Ho, Amy C Peterson, Sujit Janardhan, Ian Brown, Kesavannair Praveen & Thomas F Gajewski

  2. Department of Biochemistry, The University of Alberta, Edmonton, T6G2E1, Alberta, Canada

    Stacey Stang & James C Stone

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Contributions

Y.Z. did experiments involving the cloning, transduction and expression of DGK-α and DGK-ζ, the effects of the DGK inhibitor, the generation of the dominant negative Cbl adenovirus and experiments associated with it, and participated in writing the manuscript; R.M. developed adenoviruses encoding active Ras and GFP and adenoviral transduction protocols, immunized mice and derived CAR transgenic TH1 clones, and showed that active Ras could restore IL-2 production by anergic TH1 cells; A.W.H. did gene expression profiling of anergic TH1 cells and confirmed the effects of active Ras on anergic T cells and effects of the DGK inhibitor; A.C.P. did all experiments with CrkL-deficient T cells; S.J. and I.B. worked together to accomplish the in vivo anergy experiments with 2C Rag2−/− T cells; K.P. did confocal microscopy of 2C cells conjugated with P815.B71 cells; S.S. and J.C.S. did immunoblot analysis of DGK-α and provided experimental guidance; T.F.G. conceived of and directed the project, oversaw all experiments, secured funding and actively participated in manuscript writing.

Corresponding author

Correspondence to Thomas F Gajewski.

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Supplementary information

Supplementary Fig. 1

Recovery of IL-2 production in anergic TH1 cells does not require massive overexpression of constitutively active Ras. (PDF 107 kb)

Supplementary Fig. 2

CrkL and T cell anergy. (PDF 121 kb)

Supplementary Fig. 3

Expression of a dominant negative Cbl augments IL-2 production. (PDF 95 kb)

Supplementary Fig. 4

Inhibition of IL-2 production in anergic TH1 cells does not require massive overexpression of DGK-α. (PDF 118 kb)

Supplementary Methods (PDF 12 kb)

Supplementary Note (XLS 4022 kb)

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Zha, Y., Marks, R., Ho, A. et al. T cell anergy is reversed by active Ras and is regulated by diacylglycerol kinase-α. Nat Immunol 7, 1166–1173 (2006). https://doi.org/10.1038/ni1394

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