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Disruption of diacylglycerol metabolism impairs the induction of T cell anergy

Nature Immunology volume 7, pages 1174–1181 (2006)Cite this article

Abstract

Anergic T cells have altered diacylglycerol metabolism, but whether that altered metabolism has a causative function in the induction of T cell anergy is not apparent. To test the importance of diacylglycerol metabolism in T cell anergy, we manipulated diacylglycerol kinases (DGKs), which are enzymes that terminate diacylglycerol-dependent signaling. Overexpression of DGK-α resulted in a defect in T cell receptor signaling that is characteristic of anergy. We generated DGK-α-deficient mice and found that DGK-α-deficient T cells had more diacylglycerol-dependent T cell receptor signaling. In vivo anergy induction was impaired in DGK-α-deficient mice. When stimulated in anergy-producing conditions, T cells lacking DGK-α or DGK-ζ proliferated and produced interleukin 2. Pharmacological inhibition of DGK-α activity in DGK-ζ-deficient T cells that received an anergizing stimulus proliferated similarly to wild-type T cells that received CD28 costimulation and prevented anergy induction. Our findings suggest that regulation of diacylglycerol metabolism is critical in determining whether activation or anergy ensues after T cell receptor stimulation.

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Figure 1: Costimulation increases the hydrolysis of PtdIns(4,5)P2 and the generation of inositol trisphosphate but does not affect the conversion of DAG to phosphatidic acid.
Figure 2: Forced DGK-α expression in Jurkat T cells blocks TCR-induced AP-1 activity but does not affect calcium flux.
Figure 3: DGK-α deficiency does not alter the surface phenotype of peripheral CD4+ T cells.
Figure 4: DGK-α deficiency results in increased DAG-dependent TCR signaling but does not affect calcium flux.
Figure 5: Dgka−/− T cells are hyperproliferative.
Figure 6: In vivo anergy induction is impaired by DGK-α deficiency.
Figure 7: DGK deficiency decreases the requirement for costimulation.
Figure 8: DGK deficiency impairs T cell anergy.

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Acknowledgements

We thank C. Bock and D. Snyder (Transgenic and Knock-out Mouse Core Facility, Comprehensive Cancer Center, Duke University, Durham, North Carolina) for generating targeted embryonic stem clones and chimeric mice, and J. Stadanlick for critical review of the manuscript. Supported by the Huntsman Cancer Foundation (M.K.T.), the US National Institutes of Health (CA95463 to M.K.T. and R01 AI058019 to G.A.K.), the Department of Energy (DEFG0204ER63829 to M.K.T.) and the National Cancer Institute (T32CA09140 to B.A.O.).

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Author notes

  1. Benjamin A Olenchock and Rishu Guo: These authors contributed equally to this work.

Authors and Affiliations

  1. Signal Transduction Program, Abramson Family Cancer Research Institute, University of Pennsylvania School of Medicine, Philadelphia, 19104, Pennsylvania, USA

    Benjamin A Olenchock, Martha Jordan & Gary A Koretzky

  2. Department of Pediatrics, Duke University Medical Center, Durham, 27710, North Carolina, USA

    Rishu Guo, Jeffery H Carpenter & Xiao-Ping Zhong

  3. Department of Internal Medicine, Huntsman Cancer Institute, University of Utah, Salt Lake City, 84112, Utah, USA

    Matthew K Topham

  4. Department of Medicine, Division of Rheumatology, University of Pennsylvania School of Medicine, Philadelphia, 19104, Pennsylvania, USA

    Gary A Koretzky

  5. Department of Immunology, Duke University Medical Center, Durham, 27710, North Carolina, USA

    Xiao-Ping Zhong

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Contributions

B.A.O., R.G., J.H.C. and M.J. contributed experimental work and data analyses; M.K.T. contributed essential reagents; G.A.K. and X.-P.Z. supervised the studies; and all authors contributed intellectually to the project.

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Correspondence to Gary A Koretzky or Xiao-Ping Zhong.

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Supplementary information

Supplementary Fig. 1

Amounts of transcripts encoding DGK-α and DGK-ζ decreased after T cell activation. (PDF 51 kb)

Supplementary Fig. 2

Generation of Dgka-/- mice. (PDF 79 kb)

Supplementary Fig. 3

DGK-α deficiency does not affect T cell development. (PDF 80 kb)

Supplementary Fig. 4

DGK-α deficiency does not alter phenotype of peripheral CD8+ cells. (PDF 63 kb)

Supplementary Fig. 5

DGK-α deficiency increases T cell proliferation in response to anergizing stimuli. (PDF 57 kb)

Supplementary Methods (PDF 67 kb)

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Olenchock, B., Guo, R., Carpenter, J. et al. Disruption of diacylglycerol metabolism impairs the induction of T cell anergy. Nat Immunol 7, 1174–1181 (2006). https://doi.org/10.1038/ni1400

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