Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Human fetal lymphoid tissue–inducer cells are interleukin 17–producing precursors to RORC+ CD127+ natural killer–like cells

Abstract

The human body contains over 500 individual lymph nodes, yet the biology of their formation is poorly understood. Here we identify human lymphoid tissue–inducer cells (LTi cells) as lineage-negative RORC+ CD127+ cells with the functional ability to interact with mesenchymal cells through lymphotoxin and tumor necrosis factor. Human LTi cells were committed natural killer (NK) cell precursors that produced interleukin 17 (IL-17) and IL-22. In vitro, LTi cells gave rise to RORC+ CD127+ NK cells that retained the ability to produce IL-17 and IL-22. Postnatally, similar populations of LTi cell–like cells and RORC+ CD127+ NK cells were present in tonsils, and both secreted IL-17 and IL-22 but no interferon-γ. Our data indicate that lymph node organogenesis is controlled by an NK cell precursor population with adaptive immune features and demonstrate a previously unappreciated link between the innate and adaptive immune systems.

This is a preview of subscription content, access via your institution

Access options

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Figure 1: LinCD127+ cells in first-trimester mesentery and second-trimester lymph nodes.
Figure 2: LinCD45intCD127hi cells express RORC and LTi cell–associated genes.
Figure 3: The fetal lymph node–derived CD4+CD3 population does not contain LTi cells.
Figure 4: LinCD45intCD127hi cells have a functional lymphoid tissue–inducing ability.
Figure 5: LTi cells are precursors of CD127+ RORC-expressing NK cells.
Figure 6: CD127+ NK cells express RORC and produce IL-17A and IL-22.
Figure 7: Postnatal tonsils contain LTi cell–like cells that produce IL-22.
Figure 8: Activated tonsil-derived LTi cells and CD127+ NK cells produce IL-17 and IL-22 but not IFN-γ.

Similar content being viewed by others

References

  1. Drayton, D.L., Liao, S., Mounzer, R.H. & Ruddle, N.H. Lymphoid organ development: from ontogeny to neogenesis. Nat. Immunol. 7, 344–353 (2006).

    Article  CAS  PubMed  Google Scholar 

  2. Mebius, R.E. Organogenesis of lymphoid tissues. Nat. Rev. Immunol. 3, 292–303 (2003).

    Article  CAS  PubMed  Google Scholar 

  3. Mebius, R.E. et al. The fetal liver counterpart of adult common lymphoid progenitors gives rise to all lymphoid lineages, CD45+CD4+CD3 cells, as well as macrophages. J. Immunol. 166, 6593–6601 (2001).

    Article  CAS  PubMed  Google Scholar 

  4. Sun, Z. et al. Requirement for RORγ in thymocyte survival and lymphoid organ development. Science 288, 2369–2373 (2000).

    Article  CAS  Google Scholar 

  5. Kurebayashi, S. et al. Retinoid-related orphan receptor γ (RORγ) is essential for lymphoid organogenesis and controls apoptosis during thymopoiesis. Proc. Natl. Acad. Sci. USA 97, 10132–10137 (2000).

    Article  CAS  PubMed  Google Scholar 

  6. Eberl, G. et al. An essential function for the nuclear receptor RORγt in the generation of fetal lymphoid tissue inducer cells. Nat. Immunol. 5, 64–73 (2004).

    Article  CAS  PubMed  Google Scholar 

  7. Boos, M.D., Yokota, Y., Eberl, G. & Kee, B.L. Mature natural killer cell and lymphoid tissue-inducing cell development requires Id2-mediated suppression of E protein activity. J. Exp. Med. 204, 1119–1130 (2007).

    Article  CAS  PubMed  Google Scholar 

  8. Yokota, Y. et al. Development of peripheral lymphoid organs and natural killer cells depends on the helix-loop-helix inhibitor Id2. Nature 397, 702–706 (1999).

    Article  CAS  PubMed  Google Scholar 

  9. Ivanov, I.I. et al. The orphan nuclear receptor RORγt directs the differentiation program of proinflammatory IL-17+ T helper cells. Cell 126, 1121–1133 (2006).

    Article  CAS  PubMed  Google Scholar 

  10. Kyriazis, A.A. & Esterly, J.R. Development of lymphoid tissues in the human embryo and early fetus. Arch. Pathol. 90, 348–353 (1970).

    CAS  PubMed  Google Scholar 

  11. Markgraf, R., von Gaudecker, B. & Muller-Hermelink, H.K. The development of the human lymph node. Cell Tissue Res. 225, 387–413 (1982).

    Article  CAS  PubMed  Google Scholar 

  12. Spencer, J., Finn, T. & Isaacson, P.G. Human Peyer's patches: an immunohistochemical study. Gut 27, 405–410 (1986).

    Article  CAS  PubMed  Google Scholar 

  13. Spencer, J., MacDonald, T.T., Finn, T. & Isaacson, P.G. The development of gut associated lymphoid tissue in the terminal ileum of fetal human intestine. Clin. Exp. Immunol. 64, 536–543 (1986).

    CAS  PubMed Central  PubMed  Google Scholar 

  14. Mebius, R.E., Rennert, P. & Weissman, I.L. Developing lymph nodes collect CD4+CD3LTβ+ cells that can differentiate to APC, NK cells, and follicular cells but not T or B cells. Immunity 7, 493–504 (1997).

    Article  CAS  Google Scholar 

  15. Facchetti, F., Blanzuoli, L., Ungari, M., Alebardi, O. & Vermi, W. Lymph node pathology in primary combined immunodeficiency diseases. Springer Semin. Immunopathol. 19, 459–478 (1998).

    Article  CAS  PubMed  Google Scholar 

  16. Cao, X. et al. Defective lymphoid development in mice lacking expression of the common cytokine receptor γ chain. Immunity 2, 223–238 (1995).

    Article  CAS  PubMed  Google Scholar 

  17. Park, S.Y. et al. Developmental defects of lymphoid cells in Jak3 kinase-deficient mice. Immunity 3, 771–782 (1995).

    Article  CAS  Google Scholar 

  18. Yoshida, H. et al. Different cytokines induce surface lymphotoxin-αβ on IL-7 receptor-α cells that differentially engender lymph nodes and Peyer's patches. Immunity 17, 823–833 (2002).

    Article  CAS  PubMed  Google Scholar 

  19. Pahal, G.S., Jauniaux, E., Kinnon, C., Thrasher, A.J. & Rodeck, C.H. Normal development of human fetal hematopoiesis between eight and seventeen weeks' gestation. Am. J. Obstet. Gynecol. 183, 1029–1034 (2000).

    Article  CAS  PubMed  Google Scholar 

  20. Cupedo, T. et al. Initiation of cellular organization in lymph nodes is regulated by non-B cell-derived signals and is not dependent on CXC chemokine ligand 13. J. Immunol. 173, 4889–4896 (2004).

    Article  CAS  PubMed  Google Scholar 

  21. Hirose, T., Smith, R.J. & Jetten, A.M. ROR gamma: the third member of ROR/RZR orphan receptor subfamily that is highly expressed in skeletal muscle. Biochem. Biophys. Res. Commun. 205, 1976–1983 (1994).

    Article  CAS  PubMed  Google Scholar 

  22. Anker, P.S. et al. Mesenchymal stem cells in human second-trimester bone marrow, liver, lung, and spleen exhibit a similar immunophenotype but a heterogeneous multilineage differentiation potential. Haematologica 88, 845–52 (2003).

    Google Scholar 

  23. Dontje, W. et al. Delta-like1-induced Notch1 signaling regulates the human plasmacytoid dendritic cell versus T-cell lineage decision through control of GATA-3 and Spi-B. Blood 107, 2446–2452 (2006).

    Article  CAS  Google Scholar 

  24. Schotte, R. et al. The transcription factor Spi-B is expressed in plasmacytoid DC precursors and inhibits T-, B-, and NK-cell development. Blood 101, 1015–1023 (2003).

    Article  CAS  PubMed  Google Scholar 

  25. Freud, A.G. et al. A human CD34+ subset resides in lymph nodes and differentiates into CD56bright natural killer cells. Immunity 22, 295–304 (2005).

    Article  CAS  PubMed  Google Scholar 

  26. Freud, A.G. & Caligiuri, M.A. Human natural killer cell development. Immunol. Rev. 214, 56–72 (2006).

    Article  CAS  PubMed  Google Scholar 

  27. Freud, A.G. et al. Evidence for discrete stages of human natural killer cell differentiation in vivo. J. Exp. Med. 203, 1033–1043 (2006).

    Article  CAS  PubMed  Google Scholar 

  28. Nishikawa, S.I., Hashi, H., Honda, K., Fraser, S. & Yoshida, H. Inflammation, a prototype for organogenesis of the lymphopoietic/hematopoietic system. Curr. Opin. Immunol. 12, 342–345 (2000).

    Article  CAS  PubMed  Google Scholar 

  29. Jones, C.E. & Chan, K. Interleukin-17 stimulates the expression of interleukin-8, growth-related oncogene-α, and granulocyte-colony-stimulating factor by human airway epithelial cells. Am. J. Respir. Cell Mol. Biol. 26, 748–753 (2002).

    Article  CAS  Google Scholar 

  30. McAllister, F. et al. Role of IL-17A, IL-17F, and the IL-17 receptor in regulating growth-related oncogene-α and granulocyte colony-stimulating factor in bronchial epithelium: implications for airway inflammation in cystic fibrosis. J. Immunol. 175, 404–412 (2005).

    Article  CAS  PubMed  Google Scholar 

  31. Ouyang, W., Kolls, J.K. & Zheng, Y. The biological functions of T helper 17 cell effector cytokines in inflammation. Immunity 28, 454–467 (2008).

    Article  CAS  PubMed  Google Scholar 

  32. Boniface, K. et al. IL-22 inhibits epidermal differentiation and induces proinflammatory gene expression and migration of human keratinocytes. J. Immunol. 174, 3695–3702 (2005).

    Article  CAS  PubMed  Google Scholar 

  33. Sa, S.M. et al. The effects of IL-20 subfamily cytokines on reconstituted human epidermis suggest potential roles in cutaneous innate defense and pathogenic adaptive immunity in psoriasis. J. Immunol. 178, 2229–2240 (2007).

    Article  CAS  PubMed  Google Scholar 

  34. Wolk, K. et al. IL-22 increases the innate immunity of tissues. Immunity 21, 241–254 (2004).

    Article  CAS  PubMed  Google Scholar 

  35. Scandella, E. et al. Restoration of lymphoid organ integrity through the interaction of lymphoid tissue-inducer cells with stroma of the T cell zone. Nat. Immunol. 9, 667–675 (2008).

    Article  CAS  Google Scholar 

  36. Coles, M.C. et al. Role of T and NK cells and IL7/IL7r interactions during neonatal maturation of lymph nodes. Proc. Natl. Acad. Sci. USA 103, 13457–13462 (2006).

    Article  CAS  PubMed  Google Scholar 

  37. Aloisi, F. & Pujol-Borrell, R. Lymphoid neogenesis in chronic inflammatory diseases. Nat. Rev. Immunol. 6, 205–217 (2006).

    Article  CAS  PubMed  Google Scholar 

  38. Armengol, M.P. et al. Thyroid autoimmune disease: demonstration of thyroid antigen-specific B cells and recombination-activating gene expression in chemokine-containing active Intrathyroidal germinal centers. Am. J. Pathol. 159, 861–873 (2001).

    Article  CAS  PubMed  Google Scholar 

  39. Weninger, W. et al. Naive T cell Recruitment to nonlymphoid tissues: a role for endothelium-expressed CC chemokine ligand 21 in autoimmune disease and lymphoid neogenesis. J. Immunol. 170, 4638–4648 (2003).

    Article  CAS  Google Scholar 

  40. Luci, C. et al. Differential roles of the transcription factor RORγt in the development of NKp46+ lymphoid tissue–inducer–like cells and NKp46 natural killer cells in gut and skin. Nat. Immunol. advance online publication, 10.1038/ni1681 (23 November 2008).

  41. Sanos, S.L. et al. RORγt and commensal microflora are required for the differentiation of mucosal interleukin 22–producing NKp46+ cells. Nat. Immunol. advance online publication, 10.1038/ni1684 (23 November 2008).

  42. Boehm, T. & Bleul, C.C. The evolutionary history of lymphoid organs. Nat. Immunol. 8, 131–135 (2007).

    Article  CAS  PubMed  Google Scholar 

  43. de Rie, M.A., Zeijlemaker, W.P. & von dem Borne, A.E. Inhibition, by vinca alkaloids and colchicine, of antigenic modulation induced by anti-CD19 monoclonal antibodies. Leuk. Res. 12, 135–141 (1988).

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank the staff of the Centers for Contraception, Sexual Health and Abortion in Leiden and Rotterdam and the Bloemenhove clinic in Heemstede for collection of fetal tissues; Y. Simons-Oosterhuis for help with quantitative PCR; M. Kuijf for acquiring tonsil tissue; and J. Samsom, R. Mebius, L. Lanier, L. Meyaard and members of the Hematology department of the Erasmus University Medical Center for critical reading of the manuscript. Supported by The Netherlands Organization for Scientific Research (VENI grant 916.66.018 to T.C.).

Author information

Authors and Affiliations

Authors

Contributions

T.C. and H.S. conceived the work; T.C. designed and did experiments for Figures 1, 2, 3, 4, 5, 6, 7, analyzed results and wrote the manuscript; N.K.C. designed, did and analyzed experiments for Figure 8; N.P. did experiments for Figures 1, 2, 3, 4, 5, 6, 7; E.J.R. assisted in flow cytometry and experimental procedures; K.W. isolated fetal organs; J.L.G. constructed the LTβR-Ig fusion protein; W.E.F. provided mesenchymal stem cells; J.J.C. assisted with the design of the experiments and interpretation of the data; H.S. designed experiments, analyzed data and cowrote the manuscript; and all authors read and provided comments on the manuscript.

Corresponding author

Correspondence to Tom Cupedo.

Ethics declarations

Competing interests

N.K.C., J.L.G. and H.S. are employees of Genentech, a company that develops and markets therapeutic drugs.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–4 and Supplementary Table 1 (PDF 249 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cupedo, T., Crellin, N., Papazian, N. et al. Human fetal lymphoid tissue–inducer cells are interleukin 17–producing precursors to RORC+ CD127+ natural killer–like cells. Nat Immunol 10, 66–74 (2009). https://doi.org/10.1038/ni.1668

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ni.1668

This article is cited by

Search

Quick links

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