Figure legends
Figure 1. Both porcine pancreatic elastase (PPE) and papain induced pulmonary emphysema, and papain reproduced the asthmatic features. (A) Protocol (B) Respiratory mechanics (C) Lung tissue stained with haematoxylin and eosin (HE; 20x) or (E) Alcian blue and periodic acid-Schiff (AB–PAS; 20x) (D) Calculation of the mean linear intercept (F) Representative bronchoalveolar lavage fluid (BALF) cytospin preparations (50x); M, macrophage; E, eosinophil. (G) Differential cell counts of cytospin preparations (H) Airway responsiveness to increasing doses of methacholine measured by subtracting the baseline from each airway resistance; *p< 0.05, **p < 0.01, ***p < 0.001; n.s. not significant.
Figure 2. Papain induced an increase in the macrophage/neutrophil- and type 2 inflammation-associated cytokines; allergic airway inflammation and emphysema persisted for 5 weeks after the final papain administration. (A) Cytokines in the BALF (B) Total serum IgE (C) Neutrophil gelatinase-associated lipocalin (NGAL) in the serum and BALF (D) Protocol (E) Differential cell counts from BALF cytospins (F) Respiratory mechanics (G) Lung tissue stained with HE; 20x. (H, I) Total and papain-specific IgE and NGAL; *p< 0.05, **p < 0.01, ***p < 0.001; ND, not detected; n.s. not significant.
Figure S1. Body weights of mice in the PPE- and papain-treated groups were not significantly different from those treated with PBS control. Weights of mice treated with PPE (n = 19), papain (n = 27) or PBS (n = 20) measured on days 0, 7, 14, and 21 as in Fig. 1A; **p< 0.01; n.s. not significant with two-way repeated measures ANOVA with Holm-Bonferroni correction.
Figure S2. Papain-induced mucus hypersecretion and allergic airway inflammation associated with elevated levels of transcript for Muc5ac, Muc5b, Eotaxin1 and Eotaxin2. Quantitative RT-PCR determination of whole lung mRNA encoding Muc5ac, Muc5b, Eotaxin1 and Eotaxin2; *p < 0.05, **p < 0.01, ***p< 0.001; n.s. not significant.
Figure S3. IL-33 in serum and BALF on day 25. Concentrations of IL-33 in serum and BALF measured by ELISA; n.s. not significant.
Figure S4. Administration of poly(I:C) augmented airway inflammation and NGAL in the PPE- and papain-treated mice; allergic airway inflammation was sustained in the papain-treated group. (A) Protocol (B) Representative BALF cytospins; L, lymphocyte; N, neutrophil; E, eosinophil (50x) (C) Differential cell counts from BALF cytospins (D) Lung tissue stained with HE (20x) (E) Cytokines in BALF (F) NGAL in the serum and BALF; *p < 0.05, **p< 0.01, ***p < 0.001; n.s. not significant; ND, not detected.
Table S1. Cytokine array data. GM-CSF, Granulocyte macrophage colony-stimulating factor; IFN-γ, Interferon gamma; IL, Interleukin; KC, Keratinocyte-derived chemokine (CXCL1); MCP-1, Monocyte chemotactic protein-1 (CCL2); M-CSF, Macrophage colony-stimulating factor; RANTES, Regulated on activation, normal T cell expressed and secreted (CCL5); TNF-α, Tumour necrosis factor alpha; VEGF, Vascular endothelial growth factor. The data are presented as the mean (±SEM). n = 3 per group; ND, not detected.