Journal: Nature communications

Article Title: A fungal protease allergen provokes airway hyperresponsiveness in asthma

doi: 10.1038/ncomms7763

Figure Lengend Snippet: ( a ) Top panel: BALB/c mice were sensitized intraperitoneally with Af crude extract (20 µg) mixed with 20 mg Alum on days 0 and 14. Mice were challenged intranasally with PBS (“naïve”) or Af extract (25 µg, “acute”) on days 25, 26 and 27, or challenged intranasally with Af extract without prior sensitization (50 µg on days 0, 7 and 14 and then with 20 µg twice per week for 4 weeks) (“chronic”). All mice were sacrificed 24 hours after the last challenge. Bottom panel: Naïve mice were challenged intranasally with Af extract (25 µg) and sacrificed after four hours or overnight. Alp 1 was detected using specific antisera and DAB staining of paraffin-fixed lung sections. Sections were counterstained with hematoxylin. Images are representative of four to five mice per group (Scale bar, 50 µm; original magnification, 40x). ( b-c ) Alp 1 was detected in lysates of whole lung tissue (b) or BAL fluid (c) by immunoblotting. ( d ) Lung sections from healthy controls and subjects with asthma [with ( Af + ) and without ( Af − ) Af sensitivity] were immunostained with anti-Alp 1 antisera as above. Images are representative of 10-14 subjects/group (Scale bar, 50 µm; original magnification, 40x). ( e ) Alp 1 staining in the smooth muscle bundle was quantified using Image-pro Plus software and represented as the Alp 1 + ASM/total ASM area (* P = 0.034, *** P = 0.0001, Kruskal Wallis). ( f ) Correlation between the PC 20 FEV1 and Alp 1 staining for each subject.

Article Snippet: Total cell number was determined by hemocytometry, and differentials were determined by light microscopic examination of a Diff-Quick (Siemens)-stained cytospin (700 cells in a minimum of 15 fields at 40x magnification).

Techniques: Staining, Western Blot, Software