Limits...
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.

Show MeSH

Related in: MedlinePlus

IL-9 expression by ILC is transienta) Flow cytometry of lung cells isolated from IL9CreR26ReYFP mice challenged with papain or PBS challenged 24 h after the last papain re-challenge. Cells were stimulated with PdBU and ionomycin for 2.5 h, stained for CD4 and assessed for eYFP and intracellular IL-4, IL-5, IL-9 and IL-13 in the CD4− lymphocyte fraction. b) Flow cytometry of IL-9 and IL-13 expression in ILC (Lin− Thy1.2+), analyzed either before the last papain re-challenge or 6, 12 and 24 h after the last papain challenge (see scheme Supplementary Fig.9). c) Frequencies of IL-9+ ILC analyzed in the lung on indicated time point after papain re-challenge (0 timepoint reflects frequencies before the last papain re-challenge) ** p=0.009 and d) IL-9 cytokine concentration in the lung homogenate. ** p=0.004. Numbers in quadrants (a, b) indicate percent cells in each. Data represents at least three (a) or two (b,c,d) independent experiments with 3 mice in each experimental group (mean ±SEM in c,d).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3198843&req=5

Figure 2: IL-9 expression by ILC is transienta) Flow cytometry of lung cells isolated from IL9CreR26ReYFP mice challenged with papain or PBS challenged 24 h after the last papain re-challenge. Cells were stimulated with PdBU and ionomycin for 2.5 h, stained for CD4 and assessed for eYFP and intracellular IL-4, IL-5, IL-9 and IL-13 in the CD4− lymphocyte fraction. b) Flow cytometry of IL-9 and IL-13 expression in ILC (Lin− Thy1.2+), analyzed either before the last papain re-challenge or 6, 12 and 24 h after the last papain challenge (see scheme Supplementary Fig.9). c) Frequencies of IL-9+ ILC analyzed in the lung on indicated time point after papain re-challenge (0 timepoint reflects frequencies before the last papain re-challenge) ** p=0.009 and d) IL-9 cytokine concentration in the lung homogenate. ** p=0.004. Numbers in quadrants (a, b) indicate percent cells in each. Data represents at least three (a) or two (b,c,d) independent experiments with 3 mice in each experimental group (mean ±SEM in c,d).

Mentions: We next examined the cytokine expression profile from eYFP+ ILC by intracellular cytokine staining 24 h after the last papain challenge. Following a short 2.5h pulse with PdBU and ionomycin, all eYFP+ ILC expressed IL-13 and the majority expressed IL-5, but only a minority were producing IL-4 (Fig. 2a). However, no IL-9 expression could be recalled in these cells by intracellular cytokine staining (Fig. 2a). IL-9 expression in CD4+ T cells in vitro suggested short-lived IL-9 production. To investigate whether IL-9 expression from ILC is similarly curtailed in papain-mediated airway inflammation, we performed a kinetic analysis of cytokine secretion after papain re-challenge in wild-type C57B6 mice. The percentage of IL-9+ ILC recovered from the lungs of wild-type mice after an acute papain challenge changed rapidly over time, with peak expression observed 6 to 12 h after challenge, and a rapid decline by 24 h (Fig. 2b,c). Analysis of IL-9 protein expression at 6, 12 or 24 h after papain challenge supported the assumption of transient IL-9 expression, with peak protein expression levels observed after 12 h (Fig. 2d). ILC isolated from reporter mice showed eYFP expression co-localized with intracellular IL-9, if analysed 12 h after papain re-challenge (Supplementary Fig. 4). Thus, ILC induced in papain lung inflammation rapidly lose IL-9 protein expression in favour of IL-5 and IL-13 expression.


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)

IL-9 expression by ILC is transienta) Flow cytometry of lung cells isolated from IL9CreR26ReYFP mice challenged with papain or PBS challenged 24 h after the last papain re-challenge. Cells were stimulated with PdBU and ionomycin for 2.5 h, stained for CD4 and assessed for eYFP and intracellular IL-4, IL-5, IL-9 and IL-13 in the CD4− lymphocyte fraction. b) Flow cytometry of IL-9 and IL-13 expression in ILC (Lin− Thy1.2+), analyzed either before the last papain re-challenge or 6, 12 and 24 h after the last papain challenge (see scheme Supplementary Fig.9). c) Frequencies of IL-9+ ILC analyzed in the lung on indicated time point after papain re-challenge (0 timepoint reflects frequencies before the last papain re-challenge) ** p=0.009 and d) IL-9 cytokine concentration in the lung homogenate. ** p=0.004. Numbers in quadrants (a, b) indicate percent cells in each. Data represents at least three (a) or two (b,c,d) independent experiments with 3 mice in each experimental group (mean ±SEM in c,d).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3198843&req=5

Figure 2: IL-9 expression by ILC is transienta) Flow cytometry of lung cells isolated from IL9CreR26ReYFP mice challenged with papain or PBS challenged 24 h after the last papain re-challenge. Cells were stimulated with PdBU and ionomycin for 2.5 h, stained for CD4 and assessed for eYFP and intracellular IL-4, IL-5, IL-9 and IL-13 in the CD4− lymphocyte fraction. b) Flow cytometry of IL-9 and IL-13 expression in ILC (Lin− Thy1.2+), analyzed either before the last papain re-challenge or 6, 12 and 24 h after the last papain challenge (see scheme Supplementary Fig.9). c) Frequencies of IL-9+ ILC analyzed in the lung on indicated time point after papain re-challenge (0 timepoint reflects frequencies before the last papain re-challenge) ** p=0.009 and d) IL-9 cytokine concentration in the lung homogenate. ** p=0.004. Numbers in quadrants (a, b) indicate percent cells in each. Data represents at least three (a) or two (b,c,d) independent experiments with 3 mice in each experimental group (mean ±SEM in c,d).
Mentions: We next examined the cytokine expression profile from eYFP+ ILC by intracellular cytokine staining 24 h after the last papain challenge. Following a short 2.5h pulse with PdBU and ionomycin, all eYFP+ ILC expressed IL-13 and the majority expressed IL-5, but only a minority were producing IL-4 (Fig. 2a). However, no IL-9 expression could be recalled in these cells by intracellular cytokine staining (Fig. 2a). IL-9 expression in CD4+ T cells in vitro suggested short-lived IL-9 production. To investigate whether IL-9 expression from ILC is similarly curtailed in papain-mediated airway inflammation, we performed a kinetic analysis of cytokine secretion after papain re-challenge in wild-type C57B6 mice. The percentage of IL-9+ ILC recovered from the lungs of wild-type mice after an acute papain challenge changed rapidly over time, with peak expression observed 6 to 12 h after challenge, and a rapid decline by 24 h (Fig. 2b,c). Analysis of IL-9 protein expression at 6, 12 or 24 h after papain challenge supported the assumption of transient IL-9 expression, with peak protein expression levels observed after 12 h (Fig. 2d). ILC isolated from reporter mice showed eYFP expression co-localized with intracellular IL-9, if analysed 12 h after papain re-challenge (Supplementary Fig. 4). Thus, ILC induced in papain lung inflammation rapidly lose IL-9 protein expression in favour of IL-5 and IL-13 expression.

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