Article Figures & Data
Figures
- FIGURE 1.
Assessment of BALF eosinophilia over time. (A) OVA model of allergic airways. (B) Cell distribution in BALF at 24 h, 3 d, and 5 d after terminal challenge. (C) Net change in eosinophil frequencies over time. Table shows difference in means at 24 h and 5 d, unpaired Student t test. One-way ANOVA with Bonferroni correction for multiple comparisons, data (mean ± SEM) are from at least two independent experiments with n = 3–5 mice per group. *p < 0.05, **p < 0.01.
- FIGURE 2.
Assessment of tissue eosinophilia over time. (A) Representative bright field images of eosinophils (red arrows indicate representative eosinophils with bright pink–stained cytoplasm) in lung tissue by Sirius Red staining (left panel, original magnification ×40; right panel, original magnification ×100) and (B) quantification of eosinophil counts at day 5. *p < 0.05. (C) Net change in eosinophil frequencies over time. Table shows difference in means at 24 h and 5 d, unpaired Student t test, #p < 0.05, **p < 0.01. (D) EAR mRNA in lung tissue at 5 d after terminal challenge. One-way ANOVA with Bonferroni correction for multiple comparisons. Data (mean ± SEM) are from at least two independent experiments with n = 3–5 mice per group. *p < 0.05.
- FIGURE 3.
Examination of eotaxins in BALF after OVA challenge. Analysis of protein concentrations by ELISA of (A) eotaxin-1 or CCL11 and (B) eotaxin-2 or CCL24. One-way ANOVA with Bonferroni correction for multiple comparisons. Data (mean ± SEM) are from at least two independent experiments with n = 3–5 mice per group. **p < 0.01, ***p < 0.001, ***p < 0.0001 compared to untreated samples within each group.
- FIGURE 4.
Evaluation of the ability of SP-A to induce eosinophil migration in vitro. (A) Migration of mouse eosinophils was measured by a plate-based assay. Migration index calculated as number of live eosinophils in the bottom chamber over control. (B) SP-A concentrations in BALF of OVA-challenged mice. One-way ANOVA with Bonferroni correction for multiple comparisons. Data (mean ± SEM) are from at least two independent experiments, with n = 2–3 replicates per treatment or 3–5 mice per group. *p < 0.05, **p < 0.01, ***p < 0.001. UT, untreated.
- FIGURE 5.
Evaluation of the ability of SP-A to induce eosinophil apoptosis in mouse and human eosinophils in vitro. (A) Time course of viability assessed by trypan blue and (B) Real-Time Cell Analyzer tracing and dose response of in vitro stimulation of mouse eosinophils by SP-A. (C) Representative flow diagrams of human eosinophil apoptosis and cell death by Annexin V and PI and quantification after 16 h incubation with SP-A. Live, Annexin V−, PI−; early apoptosis, Annexin V+, PI−; late apoptosis/dead, Annexin V+, PI+. (D) Densitometry of caspase-3 by Western blot of mouse eosinophils standardized to nontreated control. ANOVA with correction for multiple comparisons. Data (mean ± SEM) are from at least two independent experiments, with n = 2–3 replicates per treatment. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
- FIGURE 6.
Evaluation of the effect of exogenous SP-A administration on eosinophils in SP-A–deficient mice after OVA challenge. (A) Schematic of OVA challenge and SP-A rescue. (B) Representative flow diagrams of eosinophil apoptosis and cell death by Annexin V and PI. (C) Total live eosinophil counts in BALF 5 d postterminal challenge. Data (mean ± SEM) are representative of two independent experiments with n = 5 mice per group. *p < 0.05.
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