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Fig. S1. Experimental protocols for examining the role of interleukin (IL)-1β in allergic airway inflammation. (a) Standard protocol. Groups of wild-type (WT), ASC−/−, NLRP3−/− or IL-1R1−/− mice on the BALB/c background were thrice sensitized with ovalbumin (OVA) subcutaneously (s.c.) in the absence of adjuvant. Injections were administered at the back of the neck. Aerosolic OVA challenge occurred over 3 consecutive days and analysis was 5 days thereafter. (b) Anakinra application during sensitization. OVA sensitizations were administered to groups of WT BALB/c mice in the absence of adjuvant. Anakinra was given on the days depicted in the scheme. Challenge and analysis was performed as in (a). (c) Anakinra application during challenge. Allergic airway inflammation (AAI) was performed as described in (a) and (b). Anakinra was administered throughout days 25–31.

Fig. S2. Functional relevance of interleukin (IL)-1β and IL-18 within BALB/c mice. (a) Bone marrow-derived dendritic cells (BMDCs) from wild-type (WT) and ASC-deficient BALB/c mice or WT C57BL/6 mice were primed with lipopolysaccharide (LPS) (5 ng/ml) for 3 h and then stimulated with adenosine triphosphate (ATP) (5 mM) for 1 h as described previously [15]. Supernatants were collected and IL-1β levels were determined by enzyme-linked immunosorbent assay (ELISA) (eBioscience), according to the manufacturer's description. (b–d) IL-18 levels within the bronchoalveloar lavage (BAL) were measured on (b) day 28 (directly after last OVA-exposure) from WT (n = 2), ASC- (n = 2) and NLRP3-deficient mice (n = 2) or day 31 from (c) WT (n = 6), ASC- (n = 5), NLRP3-deficient mice (n = 7), (d) WT (n = 4) and IL-1R1-deficient mice (n = 10). Results show data from (a,c,d) two independent and (b) one representative experiment. Asterisks show significant differences between the indicated brackets (**P < 0·01 and ***P < 0·001).

Fig. S3. Eosinophil infiltration is reduced during airway inflammation (AAI) in ASC-, NLRP3-, interleukin (IL)-1R1-deficient and anakinra-treated mice. Mice were sensitized with either phosphate-buffered saline (PBS) or ovalbumin (OVA) (days 0, 7, 14) and subsequently challenged with OVA (days 26–28) to induce allergic airway inflammation. On day 31, total numbers of eosinophils, macrophages and neutrophils were determined within the bronchoalveolar lavage (BAL) from (a–c). Groups of wild-type (WT) (OVA: n = 27; PBS: n = 11), ASC−/− (OVA: n = 15; PBS: n = 9) or NLRP3−/− (OVA: n = 20; PBS: n = 6) and (d–f) WT (OVA: n = 11; PBS: n = 3) or IL-1R1−/− (OVA: n = 15; PBS: n = 4) mice. (g–i) In addition, groups of PBS- or OVA-treated WT mice were treated with 150 mg/kg (subcutaneously) anakinra (Kineret®) during either sensitization (+ anakinraSens) or challenge (+ anakinraChall) phases and total eosinophils, macrophages and neutrophils per BAL were obtained from PBS control groups: n = 4 (− anakinra), n = 2 (+ anakinraSens) and n = 2 (+ anakinraChall) and OVA groups: n = 7 (− anakinra), n = 8 (+ anakinraSens) and n = 8 (+ anakinraChall). Symbols show mean ± standard deviation of individual mice from (a–c) three and (d–i) two independent experiments and asterisks show significant differences between the indicated brackets (*P < 0·05).

Fig. S4. Low-dose anakinra treatment suppresses eosinophil infiltration. Groups of phosphate-buffered saline (PBS)- or ovalbumin (OVA)-treated wild-type (WT) mice were treated with 15 mg/kg (intraperitoneally) anakinra (Kineret®) during either sensitization (+ anakinraSens) or challenge (+ anakinraChall) phases. (a) Eosinophil infiltration, (b) inflammation score, (c) goblet cell infiltration, (d) cytokine responses [bronchoalveolar lavage (BAL)] and (e) OVA-specific immunoglobulin (Ig)E and IgG1 levels in sera were analysed on day 31. PBS control groups, n = 4 (− anakinra), n = 2 (+ anakinraSens) and n = 2 (+ anakinraChall) and OVA groups were n = 7 (− anakinra), n = 8 (+ anakinraSens) and n = 8 (+ anakinraChall). Symbols and bars show mean ± standard deviation of individual mice from two independent experiments and asterisks show significant differences between the brackets (**P < 0·01 and *P < 0·05).

Fig. S5. Anakinra treatment suppresses eosinophil infiltration and dampens goblet cell influx. Groups of phosphate-buffered saline (PBS)- or ovalbumin (OVA)-treated wild-type (WT) mice were treated with 100 mg/kg (intraperitoneally) anakinra (Kineret®) during either sensitization (+ anakinraSens) or challenge (+ anakinraChall) phases. (a) Eosinophil infiltration, (b) inflammation score, (c) goblet cell infiltration, (d) cytokine responses [bronchoalveolar lavage (BAL)] and (e) OVA-specific immunoglobulin (Ig)E and IgG1 levels in sera were analysed on day 31. PBS control groups, n = 4 (− anakinra), n = 2 (+ anakinraSens) and n = 2 (+ anakinraChall) and OVA groups were n = 8 (− anakinra), n = 9 (+ anakinraSens) and n = 7 (+ anakinraChall). Symbols and bars show the mean ± standard deviation of individual mice from two independent experiments and asterisks show significant differences between the brackets (**P < 0·01).

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