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Figure 1.

rCARDS toxin exacerbates inflammation associated with OVA treatment.

A-B) H&E stained lung sections demonstrating inflammation in OVA (panel A) or OVA + rCARDS toxin (panel B) 40x magnification. C) Quantification of average histological scores from OVA or OVA + rCARDS toxin mice on day 7 (p = 0.0002).

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Figure 1 Expand

Figure 2.

rCARDS toxin exacerbates eosinophilia.

A) Cellular differentials from BALF on day 7 after exposure to CF or rCARDS toxin. Increased eosinophilia is seen at day 7 (p = 0.006). B-C) Eosinophils at peribronchiolar lesions of OVA (B) or OVA + rCARDS toxin (C) detected by immunohistochemistry with anti-MBP. Brown color depicts positive staining for eosinophil MBP. Original magnification 40x. D) Quantification of bronchiolar MBP immunohistochemistry stain using Aperio digital pathology system. E) Measurement of eotaxin-1 concentration by ELISA (n.s.). F) Differences in eotaxin-2 concentrations determined by ELISA (p<0.0001). G) H&E stain depicting eosinophil-rich granulomas 40x magnification. Red arrows point towards giant cells. H) MBP immunohistochemistry of granulomatous lesion 40x magnification. Black arrows point towards eosinophils and red arrows point towards giant cells.

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Figure 3.

rCARDS toxin increases the induction of Th2 effector cytokines in OVA treated mice.

RNA was extracted from lungs of OVA- or OVA + rCARDS toxin-treated mice. qRT-PCR quantification of mRNA induction for A) IL-4 (p = 0.003) and B) IL-13 (p = 0.0316) 7 days after exposure.

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Figure 4.

Chemokines CCL17 and CCL22 expression is increased in OVA-treated mice exposed to rCARDS toxin.

qRT-PCR quantification of mRNA induction for A) CCL17 and B) CCL22 (p = 0.03) 7 days after exposure. Chemokine concentrations were measured by ELISA for C) CCL17. (p = 0.005) and D) CCL22 (p = 0.002).

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Figure 5.

rCARDS toxin exacerbates OVA-induced airway hyperreactivity in BALB/cJ mice.

Airway hyperreactivity was directly measured by changes in airway resistance and lung compliance through a methacholine dose curve. (A) Dynamic airway resistance (p<0.0001) and (B) Dynamic compliance (p = 0.0005) measurements were made 7 days after CARDS toxin exposure and analyzed using 2-way ANOVA.

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