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An IL-9 fate reporter demonstrates the induction of an innate IL-9 response in lung inflammation.

Wilhelm C, Hirota K, Stieglitz B, Van Snick J, Tolaini M, Lahl K, Sparwasser T, Helmby H, Stockinger B - Nat. Immunol. (2011)

Bottom Line: We found that during papain-induced lung inflammation, IL-9 production was largely restricted to innate lymphoid cells (ILCs).IL-9 production by ILCs depended on IL-2 from adaptive immune cells and was rapidly lost in favor of other cytokines, such as IL-13 and IL-5.Blockade of IL-9 production via neutralizing antibodies resulted in much lower expression of IL-13 and IL-5, which suggested that ILCs provide the missing link between the well-established functions of IL-9 in the regulation of type 2 helper T cell cytokines and responses.

View Article: PubMed Central - PubMed

Affiliation: Division of Molecular Immunology, Medical Research Council National Institute for Medical Research, Mill Hill, UK.

ABSTRACT
Interleukin 9 (IL-9) is a cytokine linked to lung inflammation, but its cellular origin and function remain unclear. Here we describe a reporter mouse strain designed to map the fate of cells that have activated IL-9. We found that during papain-induced lung inflammation, IL-9 production was largely restricted to innate lymphoid cells (ILCs). IL-9 production by ILCs depended on IL-2 from adaptive immune cells and was rapidly lost in favor of other cytokines, such as IL-13 and IL-5. Blockade of IL-9 production via neutralizing antibodies resulted in much lower expression of IL-13 and IL-5, which suggested that ILCs provide the missing link between the well-established functions of IL-9 in the regulation of type 2 helper T cell cytokines and responses.

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Papain induced eYFP+ cells do not express any lineage markers(a) Number of cells in the lungs of IL9CreR26ReYFP mice challenged with papain (black bars) and PBS (white bars) on days 0, 3, 6, 14 and analysed 20 h after the last re-challenge (see scheme Supplementary Fig.9). (b) Cytokine concentration in the broncheo-alveolar lavage fluid (BALF) of mice immunized with papain (black bars) or PBS (white bars) as in a. (c,d) Flow cytometry of total lung cells from papain challenged IL9CreR26ReYFP mice as in a stained for CD4 (c) or TCRβ, CD8α, TCRγδ, CD19, Nkp46, Gr-1, CD11c, SiglecF and CD11b (d) and assessed for eYFP expression. (e) Absolute number of eYFP+ ILC in the lung of papain challenged IL9CreR26ReYFP mice. g) Flow cytometry of CD45, Thy1.2, IL33R, CD25, MHC II, Sca-1 and eYFP expression in papain-challenged IL9CreR26ReYFP mice. Data represent at least three independent experiments with 3 mice in each experimental group (mean ±SEM in b, c, f). DCs, dendritic cells; Eos, Eosinophils: Mφ, Macrophages: Neu, Neutrophils.
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Figure 1: Papain induced eYFP+ cells do not express any lineage markers(a) Number of cells in the lungs of IL9CreR26ReYFP mice challenged with papain (black bars) and PBS (white bars) on days 0, 3, 6, 14 and analysed 20 h after the last re-challenge (see scheme Supplementary Fig.9). (b) Cytokine concentration in the broncheo-alveolar lavage fluid (BALF) of mice immunized with papain (black bars) or PBS (white bars) as in a. (c,d) Flow cytometry of total lung cells from papain challenged IL9CreR26ReYFP mice as in a stained for CD4 (c) or TCRβ, CD8α, TCRγδ, CD19, Nkp46, Gr-1, CD11c, SiglecF and CD11b (d) and assessed for eYFP expression. (e) Absolute number of eYFP+ ILC in the lung of papain challenged IL9CreR26ReYFP mice. g) Flow cytometry of CD45, Thy1.2, IL33R, CD25, MHC II, Sca-1 and eYFP expression in papain-challenged IL9CreR26ReYFP mice. Data represent at least three independent experiments with 3 mice in each experimental group (mean ±SEM in b, c, f). DCs, dendritic cells; Eos, Eosinophils: Mφ, Macrophages: Neu, Neutrophils.

Mentions: Because IL-9 expression has been linked to lung inflammation, we studied the IL-9 expression pattern in IL9CreR26ReYFP mice in a setting of airway inflammation. Papain, a protease with adjuvant activity that leads to asthma-like responses32 induces a strong TH2 phenotype upon subcutaneous injection33 and has been used to induce experimental airway inflammation34,35. Intranasal challenge of IL9CreR26ReYFP reporter mice (Fig. 1a) with papain induces a robust TH2-type lung inflammation as monitored by cell influx in the lungs (Fig. 1b) and cytokine production in the airways (Fig.1c). Notably, papain induced airway inflammation resulted in IL-9 expression in the lung (Fig. 1c) and analysis of reporter mice revealed that most, if not all, eYFP expression was found in non-T cells (Fig. 1d). Further phenotyping of these eYFP+ cells showed that they were not related to any defined haematopoietic lineage (Fig. 1e). No eYFP+ Lin− cells were recovered from the lungs of mice challenged with PBS as control (Fig. 1f).


An IL-9 fate reporter demonstrates the induction of an innate IL-9 response in lung inflammation.

Wilhelm C, Hirota K, Stieglitz B, Van Snick J, Tolaini M, Lahl K, Sparwasser T, Helmby H, Stockinger B - Nat. Immunol. (2011)

Papain induced eYFP+ cells do not express any lineage markers(a) Number of cells in the lungs of IL9CreR26ReYFP mice challenged with papain (black bars) and PBS (white bars) on days 0, 3, 6, 14 and analysed 20 h after the last re-challenge (see scheme Supplementary Fig.9). (b) Cytokine concentration in the broncheo-alveolar lavage fluid (BALF) of mice immunized with papain (black bars) or PBS (white bars) as in a. (c,d) Flow cytometry of total lung cells from papain challenged IL9CreR26ReYFP mice as in a stained for CD4 (c) or TCRβ, CD8α, TCRγδ, CD19, Nkp46, Gr-1, CD11c, SiglecF and CD11b (d) and assessed for eYFP expression. (e) Absolute number of eYFP+ ILC in the lung of papain challenged IL9CreR26ReYFP mice. g) Flow cytometry of CD45, Thy1.2, IL33R, CD25, MHC II, Sca-1 and eYFP expression in papain-challenged IL9CreR26ReYFP mice. Data represent at least three independent experiments with 3 mice in each experimental group (mean ±SEM in b, c, f). DCs, dendritic cells; Eos, Eosinophils: Mφ, Macrophages: Neu, Neutrophils.
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Figure 1: Papain induced eYFP+ cells do not express any lineage markers(a) Number of cells in the lungs of IL9CreR26ReYFP mice challenged with papain (black bars) and PBS (white bars) on days 0, 3, 6, 14 and analysed 20 h after the last re-challenge (see scheme Supplementary Fig.9). (b) Cytokine concentration in the broncheo-alveolar lavage fluid (BALF) of mice immunized with papain (black bars) or PBS (white bars) as in a. (c,d) Flow cytometry of total lung cells from papain challenged IL9CreR26ReYFP mice as in a stained for CD4 (c) or TCRβ, CD8α, TCRγδ, CD19, Nkp46, Gr-1, CD11c, SiglecF and CD11b (d) and assessed for eYFP expression. (e) Absolute number of eYFP+ ILC in the lung of papain challenged IL9CreR26ReYFP mice. g) Flow cytometry of CD45, Thy1.2, IL33R, CD25, MHC II, Sca-1 and eYFP expression in papain-challenged IL9CreR26ReYFP mice. Data represent at least three independent experiments with 3 mice in each experimental group (mean ±SEM in b, c, f). DCs, dendritic cells; Eos, Eosinophils: Mφ, Macrophages: Neu, Neutrophils.
Mentions: Because IL-9 expression has been linked to lung inflammation, we studied the IL-9 expression pattern in IL9CreR26ReYFP mice in a setting of airway inflammation. Papain, a protease with adjuvant activity that leads to asthma-like responses32 induces a strong TH2 phenotype upon subcutaneous injection33 and has been used to induce experimental airway inflammation34,35. Intranasal challenge of IL9CreR26ReYFP reporter mice (Fig. 1a) with papain induces a robust TH2-type lung inflammation as monitored by cell influx in the lungs (Fig. 1b) and cytokine production in the airways (Fig.1c). Notably, papain induced airway inflammation resulted in IL-9 expression in the lung (Fig. 1c) and analysis of reporter mice revealed that most, if not all, eYFP expression was found in non-T cells (Fig. 1d). Further phenotyping of these eYFP+ cells showed that they were not related to any defined haematopoietic lineage (Fig. 1e). No eYFP+ Lin− cells were recovered from the lungs of mice challenged with PBS as control (Fig. 1f).

Bottom Line: We found that during papain-induced lung inflammation, IL-9 production was largely restricted to innate lymphoid cells (ILCs).IL-9 production by ILCs depended on IL-2 from adaptive immune cells and was rapidly lost in favor of other cytokines, such as IL-13 and IL-5.Blockade of IL-9 production via neutralizing antibodies resulted in much lower expression of IL-13 and IL-5, which suggested that ILCs provide the missing link between the well-established functions of IL-9 in the regulation of type 2 helper T cell cytokines and responses.

View Article: PubMed Central - PubMed

Affiliation: Division of Molecular Immunology, Medical Research Council National Institute for Medical Research, Mill Hill, UK.

ABSTRACT
Interleukin 9 (IL-9) is a cytokine linked to lung inflammation, but its cellular origin and function remain unclear. Here we describe a reporter mouse strain designed to map the fate of cells that have activated IL-9. We found that during papain-induced lung inflammation, IL-9 production was largely restricted to innate lymphoid cells (ILCs). IL-9 production by ILCs depended on IL-2 from adaptive immune cells and was rapidly lost in favor of other cytokines, such as IL-13 and IL-5. Blockade of IL-9 production via neutralizing antibodies resulted in much lower expression of IL-13 and IL-5, which suggested that ILCs provide the missing link between the well-established functions of IL-9 in the regulation of type 2 helper T cell cytokines and responses.

Show MeSH
Related in: MedlinePlus