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Negative regulation of natural killer cell function by EAT-2, a SAP-related adaptor

Nature Immunology volume 6, pages 1002–1010 (2005)Cite this article

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Abstract

EAT-2 is an adaptor expressed in innate immune cells, including natural killer (NK) cells. It is closely related to the adaptor SAP, which regulates signaling lymphocyte activation molecule (SLAM)–related receptors by recruiting the kinase FynT to the receptors. Here we have studied the function of EAT-2 in NK cells by creating mice lacking or overexpressing EAT-2. Like SAP, EAT-2 was associated with the SLAM-related receptor 2B4 in NK cells. However, unlike SAP, EAT-2 was an inhibitor of NK cell function. EAT-2 repressed natural cytotoxicity and interferon-γ secretion by a mechanism involving tyrosine phosphorylation of its C terminus.* We have demonstrated a similar function for the adaptor ERT, a newly identified SAP family member expressed in mouse NK cells. These data identify a previously unknown mechanism of NK cell inhibition. Moreover, they indicate that EAT-2 and SAP have distinct and at times opposing functions in natural immunity.

* NOTE: In the version of this article initially published online, the sixth line of the abstract was incorrect; it should begin “…its C terminus.”

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Figure 1: Expression of SAP family adaptors in normal mouse immune cells.
Figure 2: Generation of Sh2d1b−/− and Sh2d1c−/− mice.
Figure 3: Functional characterization of IL-2-activated NK cells from EAT-2-deficient and ERT-deficient mice.
Figure 4: Effect of EAT-2 deficiency on ex vivo and in vivo NK cell function.
Figure 5: Functional analyses of NK cells from transgenic mice overexpressing EAT-2 or SAP.
Figure 6: Association of EAT-2 and ERT and of SAP with 2B4 but not with other SLAM-related receptors in mouse NK cells.
Figure 7: EAT-2 and ERT, but not SAP, are tyrosine-phosphorylated in mouse NK cells.
Figure 8: The C-terminal tyrosines of EAT-2 are required for inhibition of NK cell function.
Figure 9: EAT-2 inhibits proximal events in stimulatory NK cell receptor signaling.

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Change history

  • 02 September 2005

    text errors corrected

Notes

  1. NOTE: In the version of this article initially published online, the sixth line of the abstract was incorrect; it should begin "…its C terminus." In the Methods section, the final rat mAb to SAP was misidentified; in column 2 of the Methods section, the identification on line 11 should read "(clone 1A9; IgG)" and line 14 should begin "…but not EAT-2 or ERT…." The error has been corrected for the HTML and print versions of the article.

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Acknowledgements

We thank A. Makrigiannis and members of the Veillette laboratory for discussions, and M. Bakinowski and M.-C. Fondanèche for technical help. Supported by the Canadian Institutes of Health Research (A.V.), the National Cancer Institute of Canada (A.V. and R.R.), the CANVAC National Centre of Excellence (A.V.), the Institut National de la Santé et de la Recherche Médicale (S.L.), the Association pour la Recherche sur le Cancer, France (S.L.), the Centre National de la Recherche Scientifique, France (S.L.) and the Canada Research Chair Program (A.V.).

Author information

Authors and Affiliations

  1. Clinical Research Institute of Montréal, Montréal, H2W 1R7, Québec, Canada

    Romain Roncagalli, James E R Taylor, Shaohua Zhang, Xiaochu Shi, Riyan Chen, Mario-Ernesto Cruz-Munoz & André Veillette

  2. Program in Molecular Biology, University of Montréal, Montréal, H2W 1R7, Québec, Canada

    Romain Roncagalli

  3. International Agency for Research on Cancer, Lyon, 69372, France

    Luo Yin

  4. Unité INSERM U429, Hôpital Necker Enfants-Malades, Paris, 75015, France

    Sylvain Latour

  5. Department of Medicine, University of Montréal, Montréal, H3C 3J7, Québec, Canada

    André Veillette

  6. Department of Medicine, McGill University, Montréal, H3G 1Y6, Québec, Canada

    André Veillette

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  1. Romain Roncagalli
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Supplementary information

Supplementary Fig. 1

Detection of SAP-related proteins in mouse immune cells. (PDF 498 kb)

Supplementary Fig. 2

NK cell populations in EAT-2-deficient mice. (PDF 649 kb)

Supplementary Fig. 3

Analyses of NK functions in ERT transgenic mice. (PDF 331 kb)

Supplementary Fig. 4

Effect of EAT-2 on activating receptors in established cell lines. (PDF 522 kb)

Supplementary Table 1

Differential impact of SAP-related adaptors on natural cytotioxicity. (PDF 61 kb)

Supplementary Methods (PDF 83 kb)

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Roncagalli, R., Taylor, J., Zhang, S. et al. Negative regulation of natural killer cell function by EAT-2, a SAP-related adaptor. Nat Immunol 6, 1002–1010 (2005). https://doi.org/10.1038/ni1242

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