Identification of an IL-17-producing NK1.1(neg) iNKT cell population involved in airway neutrophilia - PubMed (original) (raw)

Comparative Study

. 2007 May 14;204(5):995-1001.

doi: 10.1084/jem.20061551. Epub 2007 Apr 30.

Affiliations

Comparative Study

Identification of an IL-17-producing NK1.1(neg) iNKT cell population involved in airway neutrophilia

Marie-Laure Michel et al. J Exp Med. 2007.

Abstract

Invariant natural killer T (iNKT) cells are an important source of both T helper type 1 (Th1) and Th2 cytokines, through which they can exert beneficial, as well as deleterious, effects in a variety of inflammatory diseases. This functional heterogeneity raises the question of how far phenotypically distinct subpopulations are responsible for such contrasting activities. In this study, we identify a particular set of iNKT cells that lack the NK1.1 marker (NK1.1(neg)) and secrete high amounts of interleukin (IL)-17 and low levels of interferon (IFN)-gamma and IL-4. NK1.1(neg) iNKT cells produce IL-17 upon synthetic (alpha-galactosylceramide [alpha-GalCer] or PBS-57), as well as natural (lipopolysaccharides or glycolipids derived from Sphingomonas wittichii and Borrelia burgdorferi), ligand stimulation. NK1.1(neg) iNKT cells are more frequent in the lung, which is consistent with a role in the natural immunity to inhaled antigens. Indeed, airway neutrophilia induced by alpha-GalCer or lipopolysaccharide instillation was significantly reduced in iNKT-cell-deficient Jalpha18(-/-) mice, which produced significantly less IL-17 in their bronchoalveolar lavage fluid than wild-type controls. Furthermore, airway neutrophilia was abolished by a single treatment with neutralizing monoclonal antibody against IL-17 before alpha-GalCer administration. Collectively, our findings reveal that NK1.1(neg) iNKT lymphocytes represent a new population of IL-17-producing cells that can contribute to neutrophil recruitment through preferential IL-17 secretion.

PubMed Disclaimer

Figures

Figure 1.

Figure 1.

iNKT cell ligands induce IL-17 production by liver MNCs. Liver MNCs from wild-type, Jα18−/−, and CD1d−/− mice were stimulated in vitro by α-GalCer (A) or synthetic B. burgdorferi glycolipids (BbGL-II [IIa–IIh]) or GalA-GSL (GSL) (C). (B) Liver MNCs from wild-type (WT) or IFN-γ−/− mice were stimulated with α-GalCer in the presence or absence of anti–IL-4 mAb. In all experiments, IL-17 levels were measured in supernatants. The addition of isotype controls did not modify IL-17 production by α-GalCer–stimulated liver MNCs and no cytokine were detected without ligand stimulation (not depicted). Data represent the mean ± the SD of two to seven individual mice. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Figure 2.

Figure 2.

NK1.1neg iNKT cells are the major iNKT subset producing IL-17. Liver MNCs from wild-type mice were stained with CD1d/α-GalCer tetramers, anti-TCRβ, and NK1.1 before sorting. (A) Representative FACS profiles obtained before (left) and after (right) sorting of CD1d/α-GalCer tetramers +NK1.1neg (NK1.1neg iNKT) and CD1d/α-GalCer tetramers +NK1.1pos (NK1.1pos iNKT) liver iNKT cells. (B–F) Sorted NK1.1neg iNKT and NK1.1pos iNKT liver MNCs were stimulated with α-GalCer (B–D) or synthetic B. burgdorferi glycolipids (BbGL-II [IIc]) or GalA-GSL (GSL; E and F) plus irradiated liver MNCs from Jα18−/− mice as APCs. Sorted CD4+CD62L+ T cells from Jα18−/− mice were stimulated with α-GalCer plus irradiated liver MNCs from Jα18−/− mice as APCs (G). 3 d later, IL-17 (B, E, and G), IL-4 (C and F), and IFN-γ (D) were measured in the supernatants. No cytokine was detected in the absence of α-GalCer stimulation, in the absence of APCs or when APCs alone were stimulated with α-GalCer (not depicted). Data represent the mean ± the SD of two to three individual experiments. *, P < 0.05. (H) Intracellular IL-17 staining was performed after in vitro stimulation of liver MNCs and analyzed among gated CD1d/α-GalCer tetramers +NK1.1neg or CD1d/α-GalCer tetramers +NK1.1pos by flow cytometry. The percentage of IL-17+ and Ig control+ cells is indicated in each graph. (I) Representative FACS profile of Vβ expression by gated NK1.1neg and NK1.1pos iNKT cells. Data (H and I) are representative of three independent experiments. nd, not detected.

Figure 3.

Figure 3.

Inhibition of IL-17 production by iNKT cells in the presence of IL-4 and IFN-γ. (A and B) Sorted liver NK1.1pos iNKT (A and F) and NK1.1neg iNKT (B) cells were cocultured with irradiated liver MNCs from Jα18−/− mice as APCs and stimulated with α-GalCer in the presence or absence of anti–IL-4 and anti–IFN-γ mAb (A) of IL-4 and IFN-γ (B), or IL-23 (F). (C–E) Sorted NK1.1pos iNKT and naive conventional T cells were cocultured with anti-CD3, anti-CD28, TGFβ, IL-1α, IL-6 and TNF-α. 3 d later, IL-17 (C), IL-4 (D), and IFN-γ (E) were measured in all supernatants. Data represent the mean ± the SD of two to three individual experiments. *, P < 0.05.

Figure 4.

Figure 4.

IL-17 production by lung MNCs stimulated with α-GaCer, PBS-57, or LPS requires iNKT cells. Total (A–C) or sorted (D) NK1.1pos iNKT and NK1.1neg iNKT cells from lung MNCs from wild-type (A–D) and Jα18−/− (A–C) mice were stimulated in vitro with α-GalCer (A–D), PBS-57 (B), or LPS (C). 3 d later, supernatants were recovered and IL-17 was measured by ELISA. Data represent the mean ± the SEM of four individual mice. No cytokine was detected without stimulation (not depicted). *, P < 0.05. (E) Representative FACS profiles showing the higher percentage of NK1.1neg iNKT cells among gated TCRβ+ iNKT cells from lung.

Figure 5.

Figure 5.

α-GalCer–, PBS-57–, or LPS-induced neutrophil recruitment to airways implicates iNKT cells. (A–D) Wild-type and Jα18−/− mice received a single i.n. dose of 10 μg LPS (A), 2 μg PBS-57 (B), or 2 μg α-GalCer (B–D) 24 h before sacrifice. The number of neutrophils recruited in BALF (A and B) and the concentration of IL-17 (C) is represented. (D) Mice were treated with anti–IL-17 mAb 24 h before i.n. exposure to α-GalCer. The number of neutrophils recruited in BALF was determined 24 h later. The injection of control mAb did not modify neutrophil recruitment (not depicted). Data represent the mean ± the SEM of 5–10 individual mice. *, P < 0.05; **, P < 0.01.

References

    1. Taniguchi, M., M. Harada, S. Kojo, T. Nakayama, and H. Wakao. 2003. The regulatory role of Vα14 NKT cells in innate and acquired immune response. Annu. Rev. Immunol. 21:483–513. - PubMed
    1. Kronenberg, M. 2005. Toward an understanding of NKT cell biology: progress and paradoxes. Annu. Rev. Immunol. 23:877–900. - PubMed
    1. Benlagha, K., D.G. Wei, J. Veiga, L. Teyton, and A. Bendelac. 2005. Characterization of the early stages of thymic NKT cell development. J. Exp. Med. 202:485–492. - PMC - PubMed
    1. Bendelac, A., P.B. Savage, and L. Teyton. 2007. The biology of NKT cells. Annu. Rev. Immunol. 25:297–336. - PubMed
    1. Kawano, T., J. Cui, Y. Koezuka, I. Toura, Y. Kaneko, K. Motoki, H. Ueno, R. Nakagawa, H. Sato, E. Kondo, et al. 1997. CD1d-restricted and TCR-mediated activation of valpha14 NKT cells by glycosylceramides. Science. 278:1626–1629. - PubMed

Publication types

MeSH terms

Substances

LinkOut - more resources