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A semi-invariant Vα10+ T cell antigen receptor defines a population of natural killer T cells with distinct glycolipid antigen–recognition properties

Abstract

Type I natural killer T cells (NKT cells) are characterized by an invariant variable region 14–joining region 18 (Vα14-Jα18) T cell antigen receptor (TCR) α-chain and recognition of the glycolipid α-galactosylceramide (α-GalCer) restricted to the antigen-presenting molecule CD1d. Here we describe a population of α-GalCer-reactive NKT cells that expressed a canonical Vα10-Jα50 TCR α-chain, which showed a preference for α-glucosylceramide (α-GlcCer) and bacterial α-glucuronic acid–containing glycolipid antigens. Structurally, despite very limited TCRα sequence identity, the Vα10 TCR–CD1d–α-GlcCer complex had a docking mode similar to that of type I TCR–CD1d–α-GalCer complexes, although differences at the antigen-binding interface accounted for the altered antigen specificity. Our findings provide new insight into the structural basis and evolution of glycolipid antigen recognition and have notable implications for the scope and immunological role of glycolipid-specific T cell responses.

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Figure 1: Identification of Jα18−/− T cells reactive to CD1d–α-GalCer.
Figure 2: Jα18−/− CD1d–α-GalCer+ NKT cells express a semi-invariant Vα10-Jα50–Vβ8+ TCR.
Figure 3: Vα10 NKT cells have a unique hierarchy of antigen recognition.
Figure 4: Vα10 NKT cells have a higher affinity for α-GlcCer and are present in wild-type mice.
Figure 5: Structural comparison of Vα10 NKT cell TCR–CD1d–α-GlcCer and type I NKT cell TCR–CD1d–α-GalCer.
Figure 6: CD1d-mediated interactions with Vα10–Vβ8.1 NKT cell TCR.
Figure 7: Lipid antigen specificity.

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References

  1. Godfrey, D.I., MacDonald, H.R., Kronenberg, M., Smyth, M.J. & Van Kaer, L. NKT cells: what's in a name? Nat. Rev. Immunol. 4, 231–237 (2004).

    CAS  Google Scholar 

  2. Cardell, S. et al. CD1-restricted CD4+ T cells in major histocompatibility complex class II-deficient mice. J. Exp. Med. 182, 993–1004 (1995).

    Article  CAS  Google Scholar 

  3. Park, S.H. et al. The mouse CD1d-restricted repertoire is dominated by a few autoreactive T cell receptor families. J. Exp. Med. 193, 893–904 (2001).

    Article  CAS  Google Scholar 

  4. Blomqvist, M. et al. Multiple tissue-specific isoforms of sulfatide activate CD1d-restricted type II NKT cells. Eur. J. Immunol. 39, 1726–1735 (2009).

    Article  CAS  Google Scholar 

  5. Cui, J.Q. et al. Requirement for Vα14 NKT cells in Il-12-mediated rejection of tumors. Science 278, 1623–1626 (1997).

    Article  CAS  Google Scholar 

  6. Renukaradhya, G.J. et al. Type I NKT cells protect (and type II NKT cells suppress) the host's innate antitumor immune response to a B cell lymphoma. Blood 111, 5637–5645 (2008).

    Article  CAS  Google Scholar 

  7. Kim, J.H., Choi, E.Y. & Chung, D.H. Donor bone marrow type II (non-Vα14Jα18 CD1d-restricted) NKT cells suppress graft-versus-host disease by producing IFN-γ and IL-4. J. Immunol. 179, 6579–6587 (2007).

    Article  CAS  Google Scholar 

  8. Terabe, M. et al. A nonclassical non- Vα14Jα18 CD1d-restricted (type II) NKT cell is sufficient for down-regulation of tumor immunosurveillance. J. Exp. Med. 202, 1627–1633 (2005).

    Article  CAS  Google Scholar 

  9. Berzins, S.P., Smyth, M.J. & Godfrey, D.I. Working with NKT cells—pitfalls and practicalities. Curr. Opin. Immunol. 17, 448–454 (2005).

    Article  CAS  Google Scholar 

  10. Gadola, S.D. et al. Structure and binding kinetics of three different human CD1d-α-galactosylceramide-specific T cell receptors. J. Exp. Med. 203, 699–710 (2006).

    Article  CAS  Google Scholar 

  11. Gadola, S.D., Dulphy, N., Salio, M. & Cerundolo, V. V alpha 24-J alpha Q-independent, CD1d-restricted recognition of alpha-galactosylceramide by human CD4+ and CD8αβ+ T lymphocytes. J. Immunol. 168, 5514–5520 (2002).

    Article  CAS  Google Scholar 

  12. Brigl, M. et al. Conserved and heterogeneous lipid antigen specificities of CD1d-restricted NKT cell receptors. J. Immunol. 176, 3625–3634 (2006).

    Article  CAS  Google Scholar 

  13. Godfrey, D.I. et al. Antigen recognition by CD1d-restricted NKT T cell receptors. Semin. Immunol. 22, 61–67 (2010).

    Article  CAS  Google Scholar 

  14. Pellicci, D.G. et al. Differential recognition of CD1d-alpha-galactosyl ceramide by the Vβ8.2 and Vβ7 semi-invariant NKT T cell receptors. Immunity 31, 47–59 (2009).

    Article  CAS  Google Scholar 

  15. Borg, N.A. et al. CD1d-lipid-antigen recognition by the semi-invariant NKT T-cell receptor. Nature 448, 44–49 (2007).

    Article  CAS  Google Scholar 

  16. Li, Y. et al. The Vα14 invariant natural killer T cell TCR forces microbial glycolipids and CD1d into a conserved binding mode. J. Exp. Med. 207, 2383–2393 (2010).

    Article  CAS  Google Scholar 

  17. Wun, K.S. et al. A molecular basis for the exquisite CD1d-restricted Ag-specificity and functional responses of NKT cells. Immunity 34, 327–339 (2011).

    Article  CAS  Google Scholar 

  18. Mallevaey, T. et al. The molecular basis of NKT cell autoreactivity and recognition of self-CD1d. Immunity 34, 315–326 (2011).

    Article  CAS  Google Scholar 

  19. Wun, K.S. et al. A minimal binding footprint on CD1d-glycolipid is a basis for selection of the unique human NKT TCR. J. Exp. Med. 205, 939–949 (2008).

    Article  CAS  Google Scholar 

  20. Scott-Browne, J.P. et al. Germline-encoded recognition of diverse glycolipids by natural killer T cells. Nat. Immunol. 8, 1105–1113 (2007).

    Article  CAS  Google Scholar 

  21. Mallevaey, T. et al. T cell receptor CDR2β and CDR3β loops collaborate functionally to shape the iNKT cell repertoire. Immunity 31, 60–71 (2009).

    Article  CAS  Google Scholar 

  22. Florence, W.C. et al. Adaptability of the semi-invariant natural killer T-cell receptor towards structurally diverse CD1d-restricted ligands. EMBO J. 28, 3579–3590 (2009).

    Article  CAS  Google Scholar 

  23. Stenstrom, M., Skold, M., Andersson, A. & Cardell, S.L. Natural killer T-cell populations in C57BL/6 and NK1.1 congenic BALB.NK mice-a novel thymic subset defined in BALB.NK mice. Immunology 114, 336–345 (2005).

    Article  Google Scholar 

  24. Yu, K.O. et al. Modulation of CD1d-restricted NKT cell responses by using N-acyl variants of α-galactosylceramides. Proc. Natl. Acad. Sci. USA 102, 3383–3388 (2005).

    Article  CAS  Google Scholar 

  25. Jervis, P.J. et al. Synthesis and biological activity of α-glucosyl C24:0 and C20:2 ceramides. Bioorg. Med. Chem. Lett. 20, 3475–3478 (2010).

    Article  CAS  Google Scholar 

  26. Wolucka, B.A., McNeil, M.R., Kalbe, L., Cocito, C. & Brennan, P.J. Isolation and characterization of a novel glucuronosyl diacylglycerol from Mycobacterium smegmatis. Biochim. Biophys. Acta 1170, 131–136 (1993).

    Article  CAS  Google Scholar 

  27. Kawano, T. et al. Cd1d-restricted and TCR-mediated activation of Vα14 NKT cells by glycosylceramides. Science 278, 1626–1629 (1997).

    Article  CAS  Google Scholar 

  28. Kinjo, Y. et al. Recognition of bacterial glycosphingolipids by natural killer T cells. Nature 434, 520–525 (2005).

    Article  CAS  Google Scholar 

  29. Mattner, J. et al. Exogenous and endogenous glycolipid antigens activate NKT cells during microbial infections. Nature 434, 525–529 (2005).

    Article  CAS  Google Scholar 

  30. Sriram, V., Du, W., Gervay-Hague, J. & Brutkiewicz, R.R. Cell wall glycosphingolipids of Sphingomonas paucimobilis are CD1d-specific ligands for NKT cells. Eur. J. Immunol. 35, 1692–1701 (2005).

    Article  CAS  Google Scholar 

  31. Zhou, D. et al. Lysosomal glycosphingolipid recognition by NKT cells. Science 306, 1786–1789 (2004).

    Article  CAS  Google Scholar 

  32. Dias, B.R. et al. Identification of iGb3 and iGb4 in melanoma B16F10-Nex2 cells and the iNKT cell-mediated antitumor effect of dendritic cells primed with iGb3. Mol. Cancer 8, 116–129 (2009).

    Article  Google Scholar 

  33. Kjer-Nielsen, L. et al. A structural basis for selection and cross-species reactivity of the semi-invariant NKT cell receptor in CD1d/glycolipid recognition. J. Exp. Med. 203, 661–673 (2006).

    Article  CAS  Google Scholar 

  34. Feng, D., Bond, C.J., Ely, L.K., Maynard, J. & Garcia, K.C. Structural evidence for a germline-encoded T cell receptor-major histocompatibility complex interaction 'codon'. Nat. Immunol. 8, 975–983 (2007).

    Article  CAS  Google Scholar 

  35. Dai, S. et al. Crossreactive T Cells spotlight the germline rules for αβ T cell-receptor interactions with MHC molecules. Immunity 28, 324–334 (2008).

    Article  CAS  Google Scholar 

  36. Godfrey, D.I., Rossjohn, J. & McCluskey, J. The fidelity, occasional promiscuity, and versatility of T cell receptor recognition. Immunity 28, 304–314 (2008).

    Article  CAS  Google Scholar 

  37. Smiley, S.T., Kaplan, M.H., Grusby, M.J. & Immunoglobulin, E. Production in the absence of interleukin-4-secreting Cd1-dependent cells. Science 275, 977–979 (1997).

    Article  CAS  Google Scholar 

  38. Sonoda, K.H., Exley, M., Snapper, S., Balk, S.P. & Stein-Streilein, J. CD1-reactive natural killer T cells are required for development of systemic tolerance through an immune-privileged site. J. Exp. Med. 190, 1215–1226 (1999).

    Article  CAS  Google Scholar 

  39. Crowe, N.Y. et al. Differential antitumor immunity mediated by NKT cell subsets in vivo. J. Exp. Med. 202, 1279–1288 (2005).

    Article  CAS  Google Scholar 

  40. Long, X. et al. Synthesis and evaluation of stimulatory properties of Sphingomonadaceae glycolipids. Nat. Chem. Biol. 3, 559–564 (2007).

    Article  CAS  Google Scholar 

  41. Casanova, J.L., Romero, P., Widmann, C., Kourilsky, P. & Maryanski, J.L. T cell receptor genes in a series of class I major histocompatibility complex-restricted cytotoxic T lymphocyte clones specific for a Plasmodium berghei nonapeptide: implications for T cell allelic exclusion and antigen-specific repertoire. J. Exp. Med. 174, 1371–1383 (1991).

    Article  CAS  Google Scholar 

  42. Day, E.B. et al. The context of epitope presentation can influence functional quality of recalled influenza A virus-specific memory CD8+ T cells. J. Immunol. 179, 2187–2194 (2007).

    Article  CAS  Google Scholar 

  43. Kedzierska, K., Turner, S.J. & Doherty, P.C. Conserved T cell receptor usage in primary and recall responses to an immunodominant influenza virus nucleoprotein epitope. Proc. Natl. Acad. Sci. USA 101, 4942–4947 (2004).

    Article  CAS  Google Scholar 

  44. Lefranc, M.P. et al. IMGT, the International ImMunoGeneTics database. Nucleic Acids Res. 26, 297–303 (1998).

    Article  CAS  Google Scholar 

  45. Garboczi, D.N. et al. Assembly, specific binding, and crystallization of a human TCR-αβ with an antigenic Tax peptide from human T lymphotropic virus type 1 and the class I MHC molecule HLA-A2. J. Immunol. 157, 5403–5410 (1996).

    CAS  PubMed  Google Scholar 

  46. Holst, J. et al. Generation of T-cell receptor retrogenic mice. Nat. Protoc. 1, 406–417 (2006).

    Article  CAS  Google Scholar 

  47. Leslie, A.G.W. Joint CCP4 + ESF-EAMCB Newsletter on Protein Crystallography. Recent changes to the MOSFLM package for processing film and image plate data 26 (1992).

  48. CCP4 (Collaborative Computational Project, 4). The CCP4 suite: Programs for protein crystallography. Acta Crystallogr. D Biol. Crystallogr. 50, 760–763 (1994).

  49. Otwinoski, Z. & Minor, W. Processing of x-ray diffraction data collected in oscillation mode. Methods Enzymol. 276, 307–326 (1997).

    Article  Google Scholar 

  50. Emsley, P. & Cowtan, K. Coot: model-building tools for molecular graphics. Acta Crystallogr. D Biol. Crystallogr. 60, 2126–2132 (2004).

    Article  Google Scholar 

  51. Zajonc, D.M. et al. Structure and function of a potent agonist for the semi-invariant natural killer T cell receptor. Nat. Immunol. 6, 810–818 (2005).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank M. Taniguchi (Chiba University Graduate School of Medicine) for Jα18−/− mice; M. Kronenberg (La Jolla Institute for Allergy and Immunology) for the baculovirus-based CD1d expression system; P. Savage (Brigham Young University) for α-GalCer (C24:1 PBS-44 analog); the Australian Synchrotron staff at the MX1 and MX2 beamlines of the Australian synchrotron for assistance with data collection; S. Mattarollo, S. Doak, S. Berzins and A. Denton for discussions and assistance with some experiments; K. Field, N. Sanders and M. Reitsma for assistance with flow cytometry; and M. Stirling and the staff of the Peter MacCallum Cancer Centre Animal House and D. Maksel from the Protein Crystallography Unit at Monash University for technical assistance. Supported by the Cancer Council of Victoria, the National Health and Medical Research Council of Australia (A.P.U., L.C.S., M.J.S. and D.I.G.), the Australian Research Council (D.I.G., O.P. and J.R.), the Cancer Research Institute (G.C.) and the US National Institutes of Health (AI45889 to S.A.P.).

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Contributions

A.P.U. identified and carried out cellular and molecular characterization of Vα10 NKT cells and produced protein complexes for crystallographic studies; O.P. solved the crystal structures and did structural analysis; G.C. and K.K. carried out studies of glycolipid specificity and function; L.C.S. did surface plasmon resonance studies; D.G.P., E.B.D., L.K.-N., J.P.V., S.J.T., G.S.B., B.C., A.G.B., S.J.W., P.I., S.A.P., J.M., M.J.S., J.R. and D.I.G. provided intellectual input and key reagents and assisted with experimental design and interpretation and writing of the manuscript; and M.J.S., J.R. and D.I.G. led the investigation together and devised the project and contributed equally to this work.

Corresponding authors

Correspondence to Jamie Rossjohn or Dale I Godfrey.

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Competing interests

S.A.P. has received payment as a consultant for Vaccinex for work related to the development of therapeutics based on glycolipids pretreated with CD1d.

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Uldrich, A., Patel, O., Cameron, G. et al. A semi-invariant Vα10+ T cell antigen receptor defines a population of natural killer T cells with distinct glycolipid antigen–recognition properties. Nat Immunol 12, 616–623 (2011). https://doi.org/10.1038/ni.2051

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