Abstract
The subclassification of immunology into innate and adaptive immunity is challenged by innate-like T lymphocytes that use innate receptors to respond rapidly to stress despite expressing T cell antigen receptors (TCRs), a hallmark of adaptive immunity. In studies that explain how such cells can straddle innate and adaptive immunity, we found that signaling via antigen receptors, whose conventional role is to facilitate clonal T cell activation, was critical for the development of innate-like T cells but then was rapidly attenuated, which accommodated the cells' innate responsiveness. These findings permitted the identification of a previously unknown innate-like T cell subset and indicate that T cell hyporesponsiveness, a state traditionally linked to tolerance, may be fundamental to T cells entering the innate compartment and thereby providing lymphoid stress surveillance.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on SpringerLink
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Medzhitov, R. & Janeway, C.A. Innate immune recognition and control of adaptive immune responses. Semin. Immunol. 10, 351–353 (1998).
Hayday, A.C. γδ T cells and the lymphoid stress-surveillance response. Immunity 31, 184–196 (2009).
Strid, J., Sobolev, O., Zafirova, B., Polic, B. & Hayday, A. The intraepithelial T cell response to NKG2D-ligands links lymphoid stress surveillance to atopy. Science 334, 1293–1297 (2011).
Jensen, K.D.C. et al. Thymic selection determines γδ T cell effector fate: antigen-naive cells make interleukin-17 and antigen-experienced cells make interferon γ. Immunity 29, 90–100 (2008).
Ribot, J.C. et al. CD27 is a thymic determinant of the balance between interferon-γ- and interleukin 17–producing γδ T cell subsets. Nat. Immunol. 10, 427–436 (2009).
Sutton, C.E. et al. Interleukin-1 and IL-23 induce innate IL-17 production from γδ T cells, amplifying Th17 responses and autoimmunity. Immunity 31, 331–341 (2009).
Lockhart, E., Green, A.M. & Flynn, J.L. IL-17 production is dominated by γδ T cells rather than CD4 T cells during Mycobacterium tuberculosis infection. J. Immunol. 177, 4662–4669 (2006).
Hamada, S. et al. IL-17A produced by γδ T cells plays a critical role in innate immunity against listeria monocytogenes infection in the liver. J. Immunol. 181, 3456–3463 (2008).
Petermann, F. et al. γδ T cells enhance autoimmunity by restraining regulatory T cell responses via an interleukin-23-dependent mechanism. Immunity 33, 351–363 (2010).
Sumaria, N. et al. Cutaneous immunosurveillance by self-renewing dermal gammadelta T cells. J. Exp. Med. 208, 505–518 (2011).
Cai, Y. et al. Pivotal role of dermal IL-17-producing γδ T cells in skin inflammation. Immunity 35, 596–610 (2011).
Michel, M.-L. et al. Interleukin 7 (IL-7) selectively promotes mouse and human IL-17-producing γδ cells. Proc. Natl. Acad. Sci. USA 109, 17549–17554 (2012).
Turchinovich, G. & Hayday, A.C. Skint-1 identifies a common molecular mechanism for the development of interferon-γ-secreting versus interleukin-17-secreting γδ T cells. Immunity 35, 59–68 (2011).
Haas, J.D. et al. CCR6 and NK1.1 distinguish between IL-17A and IFN-γ-producing γδ T cells. Eur. J. Immunol. 39, 3488–3497 (2009).
Sakaguchi, N. et al. Altered thymic T-cell selection due to a mutation of the ZAP-70 gene causes autoimmune arthritis in mice. Nature 426, 454–460 (2003).
Itohara, S. et al. Homing of a γδ thymocyte subset with homogeneous T-cell receptors to mucosal epithelia. Nature 343, 754–757 (1990).
Shibata, K. et al. Identification of CD25+ γδ T cells as fetal thymus-derived naturally occurring IL-17 producers. J. Immunol. 181, 5940–5947 (2008).
Haas, J.D. et al. Development of interleukin-17-producing γδ T cells is restricted to a functional embryonic wave. Immunity 37, 48–59 (2012).
Zikherman, J., Parameswaran, R. & Weiss, A. Endogenous antigen tunes the responsiveness of naive B cells but not T cells. Nature 489, 160–164 (2012).
Weiss, A., Imboden, J., Shoback, D. & Stobo, J. Role of T3 surface molecules in human T-cell activation: T3-dependent activation results in an increase in cytoplasmic free calcium. Proc. Natl. Acad. Sci. USA 81, 4169–4173 (1984).
Osborne, B.A. et al. Identification of genes induced during apoptosis in T lymphocytes. Immunol. Rev. 142, 301–320 (1994).
Moran, A.E. et al. T cell receptor signal strength in Treg and iNKT cell development demonstrated by a novel fluorescent reporter mouse. J. Exp. Med. 208, 1279–1289 (2011).
Jameson, J. et al. A role for skin γδ T cells in wound repair. Science 296, 747–749 (2002).
Hayday, A. & Tigelaar, R. Immunoregulation in the tissues by γδ T cells. Nat. Rev. Immunol. 3, 233–242 (2003).
Strid, J. et al. Acute upregulation of an NKG2D ligand promotes rapid reorganization of a local immune compartment with pleiotropic effects on carcinogenesis. Nat. Immunol. 9, 146–154 (2008).
Chodaczek, G., Papanna, V.V., Zal, M.A.M. & Zal, T.T. Body-barrier surveillance by epidermal γδ TCRs. Nat. Immunol. 13, 272–282 (2012).
Lewis, J.M. et al. Selection of the cutaneous intraepithelial γδ+ T cell repertoire by a thymic stromal determinant. Nat. Immunol. 7, 843–850 (2006).
Boyden, L.M. et al. Skint1, the prototype of a newly identified immunoglobulin superfamily gene cluster, positively selects epidermal γδ T cells. Nat. Genet. 40, 656–662 (2008).
Jin, Y. et al. Cutting edge: Intrinsic programming of thymic γδT cells for specific peripheral tissue localization. J. Immunol. 185, 7156–7160 (2010).
Mallick-Wood, C.A. et al. Disruption of epithelial γδ T cell repertoires by mutation of the Syk tyrosine kinase. Proc. Natl. Acad. Sci. USA 93, 9704–9709 (1996).
Witherden, D.A.D. et al. The junctional adhesion molecule JAML is a costimulatory receptor for epithelial γδ T cell activation. Science 329, 1205–1210 (2010).
Zheng, Y. et al. Egr2-dependent gene expression profiling and ChIP-Seq reveal novel biologic targets in T cell anergy. Mol. Immunol. 55, 283–291 (2013).
Greenwald, R.J., Boussiotis, V.A., Lorsbach, R.B., Abbas, A.K. & Sharpe, A.H. CTLA-4 regulates induction of anergy in vivo. Immunity 14, 145–155 (2001).
Hannier, S., Tournier, M., Bismuth, G. & Triebel, F. CD3/TCR complex-associated lymphocyte activation gene-3 molecules inhibit CD3/TCR signaling. J. Immunol. 161, 4058–4065 (1998).
Olenchock, B.A. et al. Disruption of diacylglycerol metabolism impairs the induction of T cell anergy. Nat. Immunol. 7, 1174–1181 (2006).
Müller, M.R. & Rao, A. NFAT, immunity and cancer: a transcription factor comes of age. Nat. Rev. Immunol. 10, 645–656 (2010).
Leishman, A.J. et al. Precursors of functional MHC class I- or class II-restricted CD8αα+ T cells are positively selected in the thymus by agonist self-peptides. Immunity 16, 355–364 (2002).
Pobezinsky, L.A. et al. Clonal deletion and the fate of autoreactive thymocytes that survive negative selection. Nat. Immunol. 13, 569–578 (2012).
Spits, H. & Cupedo, T. Innate lymphoid cells: emerging insights in development, lineage relationships, and function. Annu. Rev. Immunol. 30, 647–675 (2012).
Rast, J.P., Smith, L.C., Loza-Coll, M., Hibino, T. & Litman, G.W. Genomic Insights into the Immune System of the Sea Urchin. Science 314, 952–956 (2006).
Vantourout, P. & Hayday, A. Six-of-the-best: unique contributions of γδ T cells to immunology. Nat. Rev. Immunol. 13, 88–100 (2013).
Dinarello, C.A.C. The interleukin-1 family: 10 years of discovery. FASEB J. 8, 1314–1325 (1994).
Guo, L. et al. IL-1 family members and STAT activators induce cytokine production by Th2, Th17, and Th1 cells. Proc. Natl. Acad. Sci. USA 106, 13463–13468 (2009).
Chaix, J. et al. Cutting edge: Priming of NK cells by IL-18. J. Immunol. 181, 1627–1631 (2008).
Willcox, C.R.C. et al. Cytomegalovirus and tumor stress surveillance by binding of a human γδ T cell antigen receptor to endothelial protein C receptor. Nat. Immunol. 13, 872–879 (2012).
Zeng, X. et al. γδ T cells recognize a microbial encoded B cell antigen to initiate a rapid antigen-specific interleukin-17 response. Immunity 37, 524–534 (2012).
Kisielow, J., Tortola, L., Weber, J., Karjalainen, K. & Kopf, M. Evidence for the divergence of innate and adaptive T-cell precursors before commitment to the αβ and γδ lineages. Blood 118, 6591–6600 (2011).
Wong, G.W. & Zúñiga-Pflücker, J.C. γδ and αβ T cell lineage choice: resolution by a stronger sense of being. Semin. Immunol. 22, 228–236 (2010).
Weber, K.S., Miller, M.J. & Allen, P.M. Th17 cells exhibit a distinct calcium profile from Th1 and Th2 cells and have Th1-like motility and NF-AT nuclear localization. J. Immunol. 180, 1442–1450 (2008).
Wang, X. et al. Human invariant natural killer T cells acquire transient innate responsiveness via histone H4 acetylation induced by weak TCR stimulation. J. Exp. Med. 209, 987–1000 (2012).
Vahl, J.C. et al. NKT cell-TCR expression activates conventional T cells in vivo, but is largely dispensable for mature NKT cell biology. PLoS Biol. 11, e1001589 (2013).
Meresse, B. et al. Coordinated induction by IL15 of a TCR-independent NKG2D signaling pathway converts CTL into lymphokine-activated killer cells in celiac disease. Immunity 21, 357–366 (2004).
Mallick-Wood, C.A. et al. Conservation of T cell receptor conformation in epidermal γδ T cells with disrupted primary Vγ gene usage. Science 279, 1729 (1998).
Krutzik, P.O. & Nolan, G.P. Intracellular phospho-protein staining techniques for flow cytometry: monitoring single cell signaling events. Cytometry A 55, 61–70 (2003).
Acknowledgements
We thank S. Sakaguchi (Osaka University) for SKG mice; K. Hogquist (University of Minnesota) for Nur77-GFP mice; and our colleagues, including M.-L. Michel, P. Vantourout, O. Sobolev, R. Hart, M. Swamy, B. Silva-Santos, D. Pennington, G. LeClercq, P. Parker and M. Saini and the staff of the flow cytometry and biological services units of the London Research Institute and of the Peter Gorer Department of Immunobiology, King's College London, for help and discussions. Supported by Cancer Research UK, Marie Curie Actions (L.D.), the University College London MBPhD programme (R.D.M.B.) and the Wellcome Trust (A.C.H.).
Author information
Authors and Affiliations
Contributions
M.W. contributed to study design, undertook experiments, analyzed data and contributed to data interpretation and to manuscript preparation and editing; G.T. contributed to study design, undertook experiments, analyzed data and contributed to data interpretation and to manuscript preparation and editing; R.D.M.B. undertook experiments and analyzed data; L.D. undertook experiments, analyzed data and contributed to manuscript editing; A.J. undertook experiments and analyzed data; A.C. contributed to study design and analysis; and A.C.H. designed the study, contributed to data interpretation and wrote and edited the manuscript.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Integrated supplementary information
Supplementary Figure 1 SKG mice are severely depleted of IL-17A-producing γδ cells.
(a) Absolute numbers of γδ T cells (TCRδ+ CD3+) isolated from indicated organs of wild type (WT) and SKG animals; Error bars are mean ± SD for 3 to 5 experiments (n ≥ 9 per group). (b) Flow cytometry analysis of γδ T cells isolated from the dermis; error bars are mean ± SD for 2 independent experiments (n = 7 per group). (c) Absolute numbers of IL-17A-producing γδ thymocytes (TCRδ+ CD3+) isolated at different time points from WT and SKG animals and stimulated with PMA and ionomycin; results shown mean ± SD (n ≥ 7 per group). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.
Supplementary Figure 2 CD27- γδ cells are hyporesponsive to TCR stimulation.
(a) LN CD27+ γδ T cells from WT or SKG mice were assessed for intracellular Ca2+ mobilization following stimulation with anti-CD3ɛ (10 μg/mL) followed by SA crosslinking (10 μg/mL); data are representative of 3 independent experiments (n=8 per group). (b) The indicated subsets of WT thymocytes from adult mice were assessed for intracellular Ca2+ mobilization as in (a); data representative of 3 independent experiments (n=11). (c) Neonatal thymocytes from WT animals were assessed for intracellular Ca2+ mobilization as in (a) in CD45+ TCRδ+ Vγ5− CD27+ and CD45+ TCRδ+ Vγ5− CD27− 17D1+ subsets; data representative of 2 independent experiments (n = 4). (d) Cells were cultured as in fig. 3 (d); IL-17A production was assessed in TCRδ+ CD3+ CD27+ subset. (e) WT LN lymphocytes were cultured for 21h with no cytokines or with IL-1β (10 ng/ml) + IL-23 (50 ng/ml) in wells coated with IgG-control or anti-CD3ɛ (10 μg/ml) and in the presence of Cyclosporin A (80 nM) or DMSO control. Brefeldin A (10 μg/ml) was added for the final 5h and IL-17A protein production assessed by intracellular cytometry in the TCRδ+ CD3+ CD27− CD44hi subset; data representative of 2 independent experiments (n = 3 per condition).
Supplementary Figure 3 Developmental changes in the phenotypes of innate-like T cell progenitors.
(a) Flow cytometry of γδ thymocytes (gated on TCRδ+CD3+) from WT embryos at indicated embryonic gestational ages and stained for CD27 and IL-7Rα or IL-1Rα; (n=3). (b) Flow cytometry of γδ fetal thymocytes (gated on TCRδ+Vγ5+) isolated from FVB. WT or Tac embryos at E16 gestational age and stained for Ly49E/F.
Supplementary Figure 4 Phenotypic traits of innate γδ T cells.
(a) Intracellular flow cytometry profile of sorted WT γδ CD27+ CD45RBhi thymocytes after stimulation with PMA + ionomycin and staining for IFN-γ; plots representative of 2 independent experiments (n = 4 per group). (b) Absolute numbers of IFN-γ-producing γδ thymocytes from adult WT or SKG mice after stimulation with PMA + ionomycin; error bars are mean ± SD; 3 independent experiments (n = 8 per group). (c) Flow cytometry of γδ T cells (gated on TCRδ+ CD3+) from LNs of WT or SKG animals stained for CD27, IL-1Rα, IL-18Rα or LAG-3; plots are representative of 2-3 independent experiments (n ≥ 6 per group); the difference in MFI for CD27 in the right panel reflects the fact that the data were collected from different independent experiments. (d) WT LN cells were cultured for 21h with indicated cytokines. Brefeldin A (10 μg/ml) was added for the final 5h and IL-17A and IFN-γ production was assessed by intracellular cytometry. Plots are gated on TCRδ+ CD3+ CD27− subset; data representative of 2 independent experiments (n = 6 per condition). (e) Expression of DGKα, DGKζ and Cbl-b was analysed by RT-PCR. Assay was performed in triplicates, and is representative of two independent experiments. *P < 0.05, **P < 0.01, *** P < 0.001.
Supplementary Figure 5 Assessment of innate-like T cell criteria in T cell subsets.
(a) Absolute numbers of γδ-NKT (TCR δ+ Vγ1+ Vδ6.3+) or iNKT (CD3+ CD1d αGal-cer tetramers+) cells in indicated organs of WT or SKG mice; error bars are mean ± SD; 2-3 independent experiments (n ≥ 4) (b) Spleen or Liver cells from Nur77-GFP reporter mice were cultured overnight with increasing amounts of coated anti-CD3ɛ (0; 0.5; 2.5 and 12.5 μg/ml). GFP expression was assessed by flow cytometry in indicated subsets; data are representative of 2 independent experiments (n ≥ 6). Data on LN γδ27+ cells are from Fig. 2c and 4d to aid comparison. (c) LN or intestinal intra-epithelial T cells from Nur77-GFP reporter mice were cultured for 4 hours on plates coated with increasing amounts of anti-CD3ɛ (0; 0.5; 2.5 and 12.5 μg/ml). GFP expression was assessed by flow cytometry in indicated subsets; data representative of 2 independent experiments (n ≥ 6).
Supplementary information
Supplementary Text and Figures
Supplementary Figures 1–5 and Supplementary Table 1 (PDF 4948 kb)
Rights and permissions
About this article
Cite this article
Wencker, M., Turchinovich, G., Di Marco Barros, R. et al. Innate-like T cells straddle innate and adaptive immunity by altering antigen-receptor responsiveness. Nat Immunol 15, 80–87 (2014). https://doi.org/10.1038/ni.2773
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/ni.2773
This article is cited by
-
Normality sensing licenses local T cells for innate-like tissue surveillance
Nature Immunology (2022)
-
Function of γδ T cells in tumor immunology and their application to cancer therapy
Experimental & Molecular Medicine (2021)
-
Ketogenesis activates metabolically protective γδ T cells in visceral adipose tissue
Nature Metabolism (2020)
-
Thymic development of unconventional T cells: how NKT cells, MAIT cells and γδ T cells emerge
Nature Reviews Immunology (2020)
-
γδ T cells compose a developmentally regulated intrauterine population and protect against vaginal candidiasis
Mucosal Immunology (2020)