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NKT cells derive from double-positive thymocytes that are positively selected by CD1d

Nature Immunology volume 2, pages 971–978 (2001)Cite this article

Abstract

CD1d-reactive NKT cells are a separate T cell sublineage. Instructive models propose that NKT cells branch off the mainstream developmental pathway because of their T cell antigen receptor specificity, whereas stochastic models would propose that they develop from precursor cells committed to this sublineage before variable-gene rearrangement. We show here that immature double-positive (DP) thymocytes form the canonical rearranged Vα gene of NKT cells at nearly equivalent frequencies in the presence or absence of CD1d expression. After interacting with CD1d in the thymus, these cells give rise to expanded populations of NKT cells—including both CD4+ and double-negative lymphocytes in the thymus and periphery—that express this α chain. These results confirm the existence of a DP intermediate for CD1d-reactive NKT cells. They also show that the early developmental stages of these T cells are not governed by a distinct mechanism, which is consistent with the TCR-instructive model of differentiation.

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Figure 1: Appearance of α-GalCer–CD1d tetramer+ cells in the thymi of C57BL/6 mice as a function of age.
Figure 2: Coreceptor expression by tetramer+ thymocytes.
Figure 3: Phenotype and proliferation of tetramer+ thymocytes during ontogeny.
Figure 4: Quantification of Vα14-Jα18 transcripts in thymocyte populations.
Figure 5: Fragment length analysis of the Vα14 rearrangements found in the different thymocyte populations of CD1d−/− and C57BL/6 mice.
Figure 6: DP TCRβ− thymocytes from C57BL/6 mice contain NKT cell precursors.
Figure 7: DP TCRβ− thymocytes from CD1d−/− mice contain NK T cell precursors.

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Acknowledgements

We thank C. Lena (FACS facility at LIAI) and A. Saluk (FACS facility at TSRI) for help with cell sorting; K.Warren for help with spectratyping; O. Naidenko and S. Sidobre for providing the α-GalCer–CD1d tetramer; and S. Hedrick, F. Koning and A. Attinger for critical review of the manuscript. Supported by grants from the National Institutes of Health (RO1 CA52511), the Human Frontiers of Science Organization (to M. K.) and the Cancer Research Institute (to L. G.).

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  1. Laurent Gapin and Jennifer L. Matsuda: These authors contributed equally to this work.

Authors and Affiliations

  1. La Jolla Institute for Allergy and Immunology, 10355 Science Center Drive, San Diego, 92121, CA, USA

    Laurent Gapin, Jennifer L. Matsuda & Mitchell Kronenberg

  2. Department of Biology, University of California at San Diego, San Diego, 92093, CA, USA

    Jennifer L. Matsuda & Mitchell Kronenberg

  3. Department of Immunology, IMM26 The Scripps Research Institute, La Jolla, 92037, CA, USA

    Charles D. Surh

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Correspondence to Mitchell Kronenberg.

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Gapin, L., Matsuda, J., Surh, C. et al. NKT cells derive from double-positive thymocytes that are positively selected by CD1d. Nat Immunol 2, 971–978 (2001). https://doi.org/10.1038/ni710

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