Enhanced T cell responses due to diacylglycerol kinase ζ deficiency

Abstract

Much is known about how T cell receptor (TCR) engagement leads to T cell activation; however, the mechanisms terminating TCR signaling remain less clear. Diacylglycerol, generated after TCR ligation, is essential in T cells. Its function must be controlled tightly to maintain normal T cell homeostasis. Previous studies have shown that diacylglycerol kinase ζ (DGKζ), which converts diacylglycerol to phosphatidic acid, can inhibit TCR signaling. Here we show that DGKζ-deficient T cells are hyperresponsive to TCR stimulation both ex vivo and in vivo. Furthermore, DGKζ-deficient mice mounted a more robust immune response to lymphocytic choriomeningitis virus infection than did wild-type mice. These results demonstrate the importance of DGKζ as a physiological negative regulator of TCR signaling and T cell activation.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: Generation of DGKζ-deficient mice by homologous recombination.
Figure 2: Immune system development.
Figure 3: Effect of DGKζ deficiency on phosphatidic acid production in T cells.
Figure 4: Enhanced Ras-ERK activation in DGKζ-deficient T cells after TCR engagement.
Figure 5: Enhanced up-regulation of activation markers in DGKζ-deficient T cells.
Figure 6: DGKζ-deficient T cells are hyperproliferative in response to TCR stimulation.
Figure 7: Enhanced homeostatic proliferation of DGKζ-deficient T cells.
Figure 8: Enhanced cellular immune responses to LCMV infection in DGKζ-deficient mice.

References

  1. 1

    Topham, M.K. & Prescott, S.M. Mammalian diacylglycerol kinases, a family of lipid kinases with signaling functions. J. Biol. Chem. 274, 11447–11450 (1999).

    CAS  Article  Google Scholar 

  2. 2

    van Blitterswijk, W.J. & Houssa, B. Properties and functions of diacylglycerol kinases. Cell. Signal. 12, 595–605 (2000).

    CAS  Article  Google Scholar 

  3. 3

    Kazanietz, M.G. Novel “nonkinase” phorbol ester receptors: the C1 domain connection. Mol. Pharmacol. 61, 759–767 (2002).

    CAS  Article  Google Scholar 

  4. 4

    Dustin, M.L. & Chan, A.C. Signaling takes shape in the immune system. Cell 103, 283–294 (2000).

    CAS  Article  Google Scholar 

  5. 5

    Kane, L.P., Lin, J. & Weiss, A. Signal transduction by the TCR for antigen. Curr. Opin. Immunol. 12, 242–249 (2000).

    CAS  Article  Google Scholar 

  6. 6

    Koretzky, G.A. & Myung, P.S. Positive and negative regulation of T-cell activation by adaptor proteins. Nat. Rev. Immunol. 1, 95–107. (2001).

    CAS  Article  Google Scholar 

  7. 7

    Yablonski, D., Kuhne, M.R., Kadlecek, T. & Weiss, A. Uncoupling of nonreceptor tyrosine kinases from PLC-γ1 in an SLP-76-deficient T cell. Science 281, 413–416 (1998).

    CAS  Article  Google Scholar 

  8. 8

    Zhang, W. et al. Association of Grb2, Gads, and phospholipase C-γ1 with phosphorylated LAT tyrosine residues. Effect of LAT tyrosine mutations on T cell angigen receptor-mediated signaling. J. Biol. Chem. 275, 23355–23361 (2000).

    CAS  Article  Google Scholar 

  9. 9

    Berridge, M.J., Heslop, J.P., Irvine, R.F. & Brown, K.D. Inositol trisphosphate formation and calcium mobilization in Swiss 3T3 cells in response to platelet-derived growth factor. Biochem. J. 222, 195–201 (1984).

    CAS  Article  Google Scholar 

  10. 10

    Crabtree, G.R. Generic signals and specific outcomes: signaling through Ca2+, calcineurin, and NF-AT. Cell 96, 611–614 (1999).

    CAS  Article  Google Scholar 

  11. 11

    Tognon, C.E. et al. Regulation of RasGRP via a phorbol ester-responsive C1 domain. Mol. Cell. Biol. 18, 6995–7008 (1998).

    CAS  Article  Google Scholar 

  12. 12

    Isakov, N. & Altman, A. Protein kinase C θ in T cell activation. Annu. Rev. Immunol. 20, 761–794 (2002).

    CAS  Article  Google Scholar 

  13. 13

    Ebinu, J.O. et al. RasGRP links T-cell receptor signaling to Ras. Blood 95, 3199–3203 (2000).

    CAS  PubMed  Google Scholar 

  14. 14

    Dower, N.A. et al. RasGRP is essential for mouse thymocyte differentiation and TCR signaling. Nat. Immunol. 1, 317–321 (2000).

    CAS  Article  Google Scholar 

  15. 15

    Coudronniere, N., Villalba, M., Englund, N. & Altman, A. NF-κB activation induced by T cell receptor/CD28 costimulation is mediated by protein kinase C-θ. Proc. Natl. Acad. Sci. USA 97, 3394–3399 (2000).

    CAS  PubMed  Google Scholar 

  16. 16

    Sun, Z. et al. PKC-θ is required for TCR-induced NF-κB activation in mature but not immature T lymphocytes. Nature 404, 402–407 (2000).

    CAS  Article  Google Scholar 

  17. 17

    Irvin, B.J., Williams, B.L., Nilson, A.E., Maynor, H.O. & Abraham, R.T. Pleiotropic contributions of phospholipase C-γ1 (PLC-γ1) to T-cell antigen receptor-mediated signaling: reconstitution studies of a PLC-γ1-deficient Jurkat T-cell line. Mol. Cell. Biol. 20, 9149–91461 (2000).

    CAS  Article  Google Scholar 

  18. 18

    Krawczyk, C. et al. Cbl-b is a negative regulator of receptor clustering and raft aggregation in T cells. Immunity 13, 463–473 (2000).

    CAS  Article  Google Scholar 

  19. 19

    Chiang, Y.J. et al. Cbl-b regulates the CD28 dependence of T-cell activation. Nature 403, 216–220 (2000).

    CAS  Article  Google Scholar 

  20. 20

    Tivol, E.A. et al. Loss of CTLA-4 leads to massive lymphoproliferation and fatal multiorgan tissue destruction, revealing a critical negative regulatory role of CTLA-4. Immunity 3, 541–547 (1995).

    CAS  Article  Google Scholar 

  21. 21

    Waterhouse, P. et al. Lymphoproliferative disorders with early lethality in mice deficient in CTLA-4. Science 270, 985–988 (1995).

    CAS  Article  Google Scholar 

  22. 22

    Di Cristofano, A. et al. Impaired Fas response and autoimmunity in Pten+/− mice. Science 285, 2122–2125 (1999).

    CAS  Article  Google Scholar 

  23. 23

    Suzuki, A. et al. T cell-specific loss of Pten leads to defects in central and peripheral tolerance. Immunity 14, 523–534 (2001).

    CAS  Article  Google Scholar 

  24. 24

    Goto, K. et al. Gene cloning, sequence, expression and in situ localization of 80 kDa diacylglycerol kinase specific to oligodendrocyte of rat brain. Brain Res. Mol. Brain Res. 16, 75–87 (1992).

    CAS  Article  Google Scholar 

  25. 25

    Sanjuan, M.A. et al. T cell activation in vivo targets diacylglycerol kinase α to the membrane: a novel mechanism for Ras attenuation. J. Immunol. 170, 2877–2883 (2003).

    CAS  Article  Google Scholar 

  26. 26

    Bunting, M., Tang, W., Zimmerman, G.A., McIntyre, T.M. & Prescott, S.M. Molecular cloning and characterization of a novel human diacylglycerol kinase ζ. J. Biol. Chem. 271, 10230–10236 (1996).

    CAS  Article  Google Scholar 

  27. 27

    Goto, K. & Kondo, H.A 104-kDa diacylglycerol kinase containing ankyrin-like repeats localizes in the cell nucleus. Proc. Natl. Acad. Sci. USA 93, 11196–11201 (1996).

    CAS  Article  Google Scholar 

  28. 28

    Zhong, X.P. et al. Regulation of T cell receptor-induced activation of the Ras-ERK pathway by diacylglycerol kinase ζ. J. Biol. Chem. 277, 31089–31098 (2002).

    CAS  Article  Google Scholar 

  29. 29

    Liu, Z., Chang, G.Q. & Leibowitz, S.F. Diacylglycerol kinase ζ in hypothalamus interacts with long form leptin receptor. Relation to dietary fat and body weight regulation. J. Biol. Chem. 276, 5900–5907 (2001).

    CAS  Article  Google Scholar 

  30. 30

    Sanjuan, M.A., Jones, D.R., Izquierdo, M. & Merida, I. Role of diacylglycerol kinase α in the attenuation of receptor signaling. J. Cell Biol. 153, 207–220 (2001).

    CAS  Article  Google Scholar 

  31. 31

    Ebinu, J.O. et al. RasGRP, a Ras guanyl nucleotide-releasing protein with calcium- and diacylglycerol-binding motifs. Science 280, 1082–1086 (1998).

    CAS  Article  Google Scholar 

  32. 32

    Rincon, M. MAP-kinase signaling pathways in T cells. Curr. Opin. Immunol. 13, 339–345 (2001).

    CAS  Article  Google Scholar 

  33. 33

    Dong, C., Davis, R.J. & Flavell, R.A. Map kinases in the immune response. Annu. Rev. Immunol. 20, 55–72 (2002).

    CAS  Article  Google Scholar 

  34. 34

    Jacinto, E., Werlen, G. & Karin, M. Cooperation between Syk and Rac1 leads to synergistic JNK activation in T lymphocytes. Immunity 8, 31–41 (1998).

    CAS  Article  Google Scholar 

  35. 35

    Cao, Y. et al. Pleiotropic defects in TCR signaling in a Vav-1-null Jurkat T-cell line. Embo J. 21, 4809–4819 (2002).

    CAS  Article  Google Scholar 

  36. 36

    Kaminuma, O., Deckert, M., Elly, C., Liu, Y.C. & Altman, A. Vav-Rac1-mediated activation of the c-Jun N-terminal kinase/c-Jun/AP-1 pathway plays a major role in stimulation of the distal NFAT site in the interleukin-2 gene promoter. Mol. Cell. Biol. 21, 3126–3136 (2001).

    CAS  Article  Google Scholar 

  37. 37

    Park, D.J., Min, H.K. & Rhee, S.G. Inhibition of CD3-linked phospholipase C by phorbol ester and by cAMP is associated with decreased phosphotyrosine and increased phosphoserine contents of PLC-γ1. J. Biol. Chem. 267, 1496–1501 (1992).

    CAS  PubMed  Google Scholar 

  38. 38

    D'Ambrosio, D., Cantrell, D.A., Frati, L., Santoni, A. & Testi, R. Involvement of p21ras activation in T cell CD69 expression. Eur. J. Immunol. 24, 616–620 (1994).

    CAS  Article  Google Scholar 

  39. 39

    Castellanos, M.C. et al. Expression of the leukocyte early activation antigen CD69 is regulated by the transcription factor AP-1. J. Immunol. 159, 5463–5473 (1997).

    CAS  PubMed  Google Scholar 

  40. 40

    Marrack, P. et al. Homeostasis of αβ TCR+ T cells. Nat. Immunol. 1, 107–111 (2000).

    CAS  Article  Google Scholar 

  41. 41

    Jung, S. & Littman, D.R. Chemokine receptors in lymphoid organ homeostasis. Curr. Opin. Immunol. 11, 319–325 (1999).

    CAS  Article  Google Scholar 

  42. 42

    Kassiotis, G., Zamoyska, R. & Stockinger, B. Involvement of avidity for major histocompatibility complex in homeostasis of naive and memory T cells. J. Exp. Med. 197, 1007–1016 (2003).

    CAS  Article  Google Scholar 

  43. 43

    Murali-Krishna, K. et al. Counting antigen-specific CD8 T cells: a reevaluation of bystander activation during viral infection. Immunity 8, 177–187 (1998).

    CAS  Article  Google Scholar 

  44. 44

    Schulz, M. et al. Major histocompatibility complex-dependent T cell epitopes of lymphocytic choriomeningitis virus nucleoprotein and their protective capacity against viral disease. Eur. J. Immunol. 19, 1657–1667 (1989).

    CAS  Article  Google Scholar 

  45. 45

    Oxenius, A. et al. Presentation of endogenous viral proteins in association with major histocompatibility complex class II: on the role of intracellular compartmentalization, invariant chain and the TAP transporter system. Eur. J. Immunol. 25, 3402–3411 (1995).

    CAS  Article  Google Scholar 

  46. 46

    Aragones, J. et al. Evidence for the involvement of diacylglycerol kinase in the activation of hypoxia-inducible transcription factor 1 by low oxygen tension. J. Biol. Chem. 276, 10548–10555 (2001).

    CAS  Article  Google Scholar 

  47. 47

    Bregoli, L., Baldassare, J.J. & Raben, D.M. Nuclear diacylglycerol kinase-θ is activated in response to α-thrombin. J. Biol. Chem. 276, 23288–23295 (2001).

    CAS  Article  Google Scholar 

  48. 48

    Cutrupi, S. et al. Src-mediated activation of α-diacylglycerol kinase is required for hepatocyte growth factor-induced cell motility. EMBO J. 19, 4614–4622 (2000).

    CAS  Article  Google Scholar 

  49. 49

    Du, X., Jiang, Y., Qian, W., Lu, X. & Walsh, J.P. Fatty acids inhibit growth-factor-induced diacylglycerol kinase α activation in vascular smooth-muscle cells. Biochem. J. 357, 275–282 (2001).

    CAS  Article  Google Scholar 

  50. 50

    Hogan, A. et al. Interaction of γ 1-syntrophin with diacylglycerol kinase-ζ Regulation of nuclear localization by PDZ interactions. J. Biol. Chem. 276, 26526–26533 (2001).

    CAS  Article  Google Scholar 

  51. 51

    Houssa, B., de Widt, J., Kranenburg, O., Moolenaar, W.H. & van Blitterswijk, W.J. Diacylglycerol kinase θ binds to and is negatively regulated by active RhoA. J. Biol. Chem. 274, 6820–6822 (1999).

    CAS  Article  Google Scholar 

  52. 52

    Inui, H., Kondo, T., Konishi, F., Kitami, Y. & Inagami, T. Participation of diacylglycerol kinase in mitogenic signal transduction induced by platelet-derived growth factor in vascular smooth muscle cells. Biochem. Biophys. Res. Commun. 205, 1338–1344 (1994).

    CAS  Article  Google Scholar 

  53. 53

    Miller, K.G., Emerson, M.D. & Rand, J.B. Goα and diacylglycerol kinase negatively regulate the Gqα pathway in C. elegans. Neuron 24, 323–333 (1999).

    CAS  Article  Google Scholar 

  54. 54

    Nagaya, H., Wada, I., Jia, Y.J. & Kanoh, H. Diacylglycerol kinase δ suppresses ER-to-Golgi traffic via its SAM and PH domains. Mol. Biol. Cell. 13, 302–316 (2002).

    CAS  Article  Google Scholar 

  55. 55

    Rodriguez de Turco, E.B. et al. Diacylglycerol kinase ε regulates seizure susceptibility and long-term potentiation through arachidonoyl-inositol lipid signaling. Proc. Natl. Acad. Sci. USA 98, 4740–4745 (2001).

    CAS  Article  Google Scholar 

  56. 56

    Topham, M.K. et al. Protein kinase C regulates the nuclear localization of diacylglycerol kinase-ζ. Nature 394, 697–700 (1998).

    CAS  Article  Google Scholar 

  57. 57

    Topham, M.K. & Prescott, S.M. Diacylglycerol kinase ζ regulates Ras activation by a novel mechanism. J. Cell Biol. 152, 1135–1143 (2001).

    CAS  Article  Google Scholar 

  58. 58

    Flores, I. et al. Diacylglycerol kinase inhibition prevents IL-2-induced G1 to S transition through a phosphatidylinositol-3 kinase-independent mechanism. J. Immunol. 163, 708–714 (1999).

    CAS  PubMed  Google Scholar 

  59. 59

    Tabellini G. et al. Diacylglycerol kinase-θ is localized in the speckle domains of the nucleus. Exp. Cell Res. 287,143–154 (2003).

    CAS  Article  Google Scholar 

  60. 60

    Schaeffer, E.M. et al. Requirement for Tec kinases Rlk and Itk in T cell receptor signaling and immunity. Science 284, 638–641 (1999).

    CAS  Article  Google Scholar 

  61. 61

    Ji, Q.-s. et al. Essential role of the tyrosine kinase substrate phospholipase C-γ1 in mammalian growth and development. Proc. Natl. Acad. Sci. USA 94, 2999–3003 (1997).

    CAS  Article  Google Scholar 

  62. 62

    Wang, D. et al. Phospholipase Cγ2 is essential in the functions of B cell and several Fc receptors. Immunity 13, 25–35 (2000).

    Article  Google Scholar 

  63. 63

    Hashimoto, K., Miyata, M., Watanabe, M. & Kano, M. Roles of phospholipase Cβ4 in synapse elimination and plasticity in developing and mature cerebellum. Mol. Neurobiol. 23, 69–82 (2001).

    CAS  Article  Google Scholar 

  64. 64

    Martin, P. et al. Role of ζ PKC in B-cell signaling and function. Embo J. 21, 4049–4057 (2002).

    CAS  Article  Google Scholar 

  65. 65

    Abeliovich, A. et al. PKC γ mutant mice exhibit mild deficits in spatial and contextual learning. Cell 75, 1263–1271 (1993).

    CAS  Article  Google Scholar 

  66. 66

    Su, T.T. et al. PKC-β controls IκB kinase lipid raft recruitment and activation in response to BCR signaling. Nat. Immunol. 3, 780–786 (2002).

    CAS  Article  Google Scholar 

  67. 67

    Masai, I., Okazaki, A., Hosoya, T. & Hotta, Y. Drosophila retinal degeneration A gene encodes an eye-specific diacylglycerol kinase with cysteine-rich zinc-finger motifs and ankyrin repeats. Proc. Natl. Acad. Sci. USA 90, 11157–11161 (1993).

    CAS  Article  Google Scholar 

  68. 68

    Newton, A.C. Protein kinase C. Seeing two domains. Curr. Biol. 5, 973–976 (1995).

    CAS  Article  Google Scholar 

  69. 69

    English, D. Phosphatidic acid: a lipid messenger involved in intracellular and extracellular signalling. Cell. Signal. 8, 341–347 (1996).

    CAS  Article  Google Scholar 

  70. 70

    Acuto O. & Cantrell, D. T cell activation and the cytoskeleton. Annu. Rev. Immunol. 18, 165–184 (2000).

    CAS  Article  Google Scholar 

  71. 71

    Ohashi, P.S. T-cell signalling and autoimmunity: molecular mechanisms of disease. Nat. Rev. Immunol. 2, 427–438 (2002).

    CAS  Article  Google Scholar 

  72. 72

    Shultz, L.D., Rajan, T.V. & Greiner, D.L. Severe defects in immunity and hematopoiesis caused by SHP-1 protein-tyrosine-phosphatase deficiency. Trends Biotechnol. 15, 302–307 (1997).

    CAS  Article  Google Scholar 

  73. 73

    Lynch, D.H., Ramsdell, F. & Alderson, M.R. Fas and FasL in the homeostatic regulation of immune responses. Immunol. Today 16, 569–74 (1995).

    CAS  Article  Google Scholar 

  74. 74

    Bird, J.J. et al. Helper T cell differentiation is controlled by the cell cycle. Immunity 9, 229–237 (1998).

    CAS  Article  Google Scholar 

  75. 75

    Ahmed, R., Salmi, A., Butler, L.D., Chiller, J.M. & Oldstone, M.B. Selection of genetic variants of lymphocytic choriomeningitis virus in spleens of persistently infected mice. Role in suppression of cytotoxic T lymphocyte response and viral persistence. J. Exp. Med. 160, 521–540 (1984).

    CAS  Article  Google Scholar 

Download references

Acknowledgements

We thank M. Kahn, V. Shapiro, J. Wu and S. Newbrough for manuscript critique; E. Peterson for discussions; S. Prescott and M. Topham for Ab to DGKζ; E. Myers and A. Lee for technical assistance; and C. Clendenin, C. Culpepper and P. Thayer of the Transgenic Core of the Abramson Family Cancer Research Institute at the University of Pennsylvania for assistance in generation of the targeted mice. Supported in part by a grant from the National Institutes of Health (G.A.K). M.S.J. is supported by a postdoctoral fellowship from the Cancer Research Institute. J.S.M. is supported by a grant from the American Society of Transplantation.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Gary A Koretzky.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Zhong, XP., Hainey, E., Olenchock, B. et al. Enhanced T cell responses due to diacylglycerol kinase ζ deficiency. Nat Immunol 4, 882–890 (2003). https://doi.org/10.1038/ni958

Download citation

Further reading

Search

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing