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Blood, Vol. 111, Issue 6, 3070-3080, March 15, 2008
Phagocytosis-independent antimicrobial activity of mast cells by means of extracellular trap formation
Blood von Köckritz-Blickwede et al.
111: 3070
Supplemental materials for: von Kockritz-Blickwede et al
Files in this Data Supplement:
- Figure S1. Granule componets are critical for the production of MCETs (JPG, 41.3 KB)
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Immunofluorescence photographs showing viable (green) versus dead (red) MCs: (Ai) Stimulation of MCs with 100mU/ml H2O2-producing enzyme glucose oxidase results in strong production of MCETs. (Aii) Degranulated MCs after 6 h of treatment with 10 µg/ml compound 48/80. Compound 48/80 has been shown to induce degranulation of MCs (Liz GZ, Chai OH, Song CH. Inhibitory effects of epigallocatechin gallate on compound 48/80-induced mast cell activation and passive cutaneous anaphylaxis. Exp Mol Med. 2005;31:290-196.). Notice that degranulation per se does not induce MCETs formation. (Aiii) Compound 48/80-treated degranulated MCs (1 h, 10 µg/ml) stimulated with H2O2-producing glucose oxidase for additional 5 h failed to produce MCETs. All bars represent 50 µm.
- Figure S2. Presence of mast cells (MCs) in biopsy samples collected from patients with streptococcal soft-tissue infections (JPG, 60.1 KB)
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Snap-frozen tissue biopsies (n=10) collected at the site of infection from patients were sectioned, fixed and immunostained for mast cell tryptase or S. pyogenes. (A) Isotype control antibody (bar, 10 µm). (B) Detection of MCs by immunohistochemical staining of human MCs tryptase (brown staining; bar, 10 µm). (C) Confocal microscopy of mast cell tryptase (stained red) and S. pyogenes (stained green) in Dapi-stained blue tissue biopsy (bar, 50 µm). (D) High magnification confocal microscopy showing colocalization of S. pyogenes and MCs tryptase (bars, 10 µm): (Di) immunostaining of S. pyogenes with Alexa green-labelled antibodies; (Dii) immunostaining of MCs tryptase with Alexa-red labelled antibodies; (Diii) overlay of A1-2 (colocalization of tryptase and bacteria results in a yellow staining (arrows)).
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