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 in ILC is dependent on IL-2 and the adaptive immune systema) Cytokine concentration in the lung homogenate of papain challenged wt (black bars) Rag1−/− (white bars) and Rag2−/−IL2rg−/− (grey bars) mice 18 h after the last papain re-challenge ***p=0.0001, **p=0.001 b) Concentration of IL-9 in the supernatant of MACS sorted ILC isolated from papain challenged wt Rag1−/− and Rag2−/−IL2rg−/− mice stimulated in vitro with IL-2 overnight. c) Cytokine concentration in the lung homogenate of papain immunized B6 mice treated with neutralising IL-2-specific (white bars) or IgG isotype control (black bars) antibody 18 h after challenge *p=0.03. d) IL-9 protein concentration in the lung homogenate of papain challenged wt (black bars) and Rag1−/− mice (white bars) analyzed 18 h after the last papain re-challenge. Where indicated papain challenged or control (PBS challenged only) Rag1−/− mice were treated intranasally with 500 ng recombinant mouse IL-2 3 h after the last papain re-challenge. e) Absolute number of ILC in the lung of papain challenged wt and papain challenged Rag1−/− mice left untreated or treated with intranasal IL-2. (treatment regimens are show in Supplementary Fig.9). Data represents two independent experiments with four mice in each experimental group (mean ±SEM).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3198843&req=5

Figure 5: IL-9 expression in ILC is dependent on IL-2 and the adaptive immune systema) Cytokine concentration in the lung homogenate of papain challenged wt (black bars) Rag1−/− (white bars) and Rag2−/−IL2rg−/− (grey bars) mice 18 h after the last papain re-challenge ***p=0.0001, **p=0.001 b) Concentration of IL-9 in the supernatant of MACS sorted ILC isolated from papain challenged wt Rag1−/− and Rag2−/−IL2rg−/− mice stimulated in vitro with IL-2 overnight. c) Cytokine concentration in the lung homogenate of papain immunized B6 mice treated with neutralising IL-2-specific (white bars) or IgG isotype control (black bars) antibody 18 h after challenge *p=0.03. d) IL-9 protein concentration in the lung homogenate of papain challenged wt (black bars) and Rag1−/− mice (white bars) analyzed 18 h after the last papain re-challenge. Where indicated papain challenged or control (PBS challenged only) Rag1−/− mice were treated intranasally with 500 ng recombinant mouse IL-2 3 h after the last papain re-challenge. e) Absolute number of ILC in the lung of papain challenged wt and papain challenged Rag1−/− mice left untreated or treated with intranasal IL-2. (treatment regimens are show in Supplementary Fig.9). Data represents two independent experiments with four mice in each experimental group (mean ±SEM).

Mentions: ILC are present in Rag1−/− mice, which lack T and B cells, but not in mice lacking the common gamma chain (IL2rg−/− mice)27-29. To investigate whether the population of IL-9-expressing cells is independent of RAG expression but dependent on signalling via the common gamma chain we analysed IL-9 expression in the lungs of papain-challenged wild-type, Rag1−/− and Rag2−/−IL2rg−/− mice. IL-9 production was reduced by 10-fold in the lungs of Rag1−/− mice as compared to wild-type mice and was absent in Rag2−/−IL2rg−/− mice (Fig. 5a). IL-5 and IL-13 protein expression was also reduced in Rag1−/− mice, in agreement with previous reports that IL-13-expressing cells in the mesenteric lymph nodes were not maintained during helminth infection in Rag1−/− mice27. However, isolation of ILC from the lungs of papain-challenged wild-type, Rag1−/− and Rag2−/−IL2rg−/− mice, followed by overnight stimulation with IL-2, resulted in similar IL-9 protein expression from ILC of wild-type and Rag1−/−(Fig. 5b). In contrast ILC isolated from Rag2−/−IL2rg−/− mice failed to produce IL-9 upon IL-2 stimulation (Fig. 5b). These results suggest that IL-9 production in ILCs upon papain challenge in vivo is dependent on the presence of adaptive immune cells, although the intrinsic capacity of ILCs to produce IL-9 is still intact in Rag1−/− mice.


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 in ILC is dependent on IL-2 and the adaptive immune systema) Cytokine concentration in the lung homogenate of papain challenged wt (black bars) Rag1−/− (white bars) and Rag2−/−IL2rg−/− (grey bars) mice 18 h after the last papain re-challenge ***p=0.0001, **p=0.001 b) Concentration of IL-9 in the supernatant of MACS sorted ILC isolated from papain challenged wt Rag1−/− and Rag2−/−IL2rg−/− mice stimulated in vitro with IL-2 overnight. c) Cytokine concentration in the lung homogenate of papain immunized B6 mice treated with neutralising IL-2-specific (white bars) or IgG isotype control (black bars) antibody 18 h after challenge *p=0.03. d) IL-9 protein concentration in the lung homogenate of papain challenged wt (black bars) and Rag1−/− mice (white bars) analyzed 18 h after the last papain re-challenge. Where indicated papain challenged or control (PBS challenged only) Rag1−/− mice were treated intranasally with 500 ng recombinant mouse IL-2 3 h after the last papain re-challenge. e) Absolute number of ILC in the lung of papain challenged wt and papain challenged Rag1−/− mice left untreated or treated with intranasal IL-2. (treatment regimens are show in Supplementary Fig.9). Data represents two independent experiments with four mice in each experimental group (mean ±SEM).
© Copyright Policy
Related In: Results  -  Collection

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

Figure 5: IL-9 expression in ILC is dependent on IL-2 and the adaptive immune systema) Cytokine concentration in the lung homogenate of papain challenged wt (black bars) Rag1−/− (white bars) and Rag2−/−IL2rg−/− (grey bars) mice 18 h after the last papain re-challenge ***p=0.0001, **p=0.001 b) Concentration of IL-9 in the supernatant of MACS sorted ILC isolated from papain challenged wt Rag1−/− and Rag2−/−IL2rg−/− mice stimulated in vitro with IL-2 overnight. c) Cytokine concentration in the lung homogenate of papain immunized B6 mice treated with neutralising IL-2-specific (white bars) or IgG isotype control (black bars) antibody 18 h after challenge *p=0.03. d) IL-9 protein concentration in the lung homogenate of papain challenged wt (black bars) and Rag1−/− mice (white bars) analyzed 18 h after the last papain re-challenge. Where indicated papain challenged or control (PBS challenged only) Rag1−/− mice were treated intranasally with 500 ng recombinant mouse IL-2 3 h after the last papain re-challenge. e) Absolute number of ILC in the lung of papain challenged wt and papain challenged Rag1−/− mice left untreated or treated with intranasal IL-2. (treatment regimens are show in Supplementary Fig.9). Data represents two independent experiments with four mice in each experimental group (mean ±SEM).
Mentions: ILC are present in Rag1−/− mice, which lack T and B cells, but not in mice lacking the common gamma chain (IL2rg−/− mice)27-29. To investigate whether the population of IL-9-expressing cells is independent of RAG expression but dependent on signalling via the common gamma chain we analysed IL-9 expression in the lungs of papain-challenged wild-type, Rag1−/− and Rag2−/−IL2rg−/− mice. IL-9 production was reduced by 10-fold in the lungs of Rag1−/− mice as compared to wild-type mice and was absent in Rag2−/−IL2rg−/− mice (Fig. 5a). IL-5 and IL-13 protein expression was also reduced in Rag1−/− mice, in agreement with previous reports that IL-13-expressing cells in the mesenteric lymph nodes were not maintained during helminth infection in Rag1−/− mice27. However, isolation of ILC from the lungs of papain-challenged wild-type, Rag1−/− and Rag2−/−IL2rg−/− mice, followed by overnight stimulation with IL-2, resulted in similar IL-9 protein expression from ILC of wild-type and Rag1−/−(Fig. 5b). In contrast ILC isolated from Rag2−/−IL2rg−/− mice failed to produce IL-9 upon IL-2 stimulation (Fig. 5b). These results suggest that IL-9 production in ILCs upon papain challenge in vivo is dependent on the presence of adaptive immune cells, although the intrinsic capacity of ILCs to produce IL-9 is still intact in Rag1−/− mice.

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