Intracellular residency of Staphylococcus aureus within mast cells in nasal polyps: A novel observation Stephen M. Hayes, MRCS, Robert Howlin, PhD, David.

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Journal of Allergy and Clinical Immunology

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Intracellular residency of Staphylococcus aureus within mast cells in nasal polyps: A novel observation Stephen M. Hayes, MRCS, Robert Howlin, PhD, David A. Johnston, PhD, Jeremy S. Webb, PhD, Stuart C. Clarke, FRCPath, FFPH, Paul Stoodley, PhD, Philip G. Harries, FRCS (ORL-HNS), Susan J. Wilson, PhD, Sylvia L.F. Pender, PhD, Saul N. Faust, PhD, FRCPCH, Luanne Hall-Stoodley, PhD, Rami J. Salib, PhD, FRCS (ORL-HNS) Journal of Allergy and Clinical Immunology Volume 135, Issue 6, Pages 1648-1651.e5 (June 2015) DOI: 10.1016/j.jaci.2014.12.1929 Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Intracellular S aureus in ex vivo NPs. Representative CLSM images of sinus mucosa and NP tissue following FISH showing Staphylococcus aureus (yellow), Staphylococcus genus biofilms (red), and bacteria hybridized only with a eubacterial probe (green). A, Aggregated bacteria are widely distributed over the epithelial surface of sinus mucosa. B, In contrast, bacterial reservoirs surround host cell nuclei in NP biopsies (white arrows). C, XYZ view: staphylococcal aggregates (white arrow) clearly visible beneath the epithelial surface (yellow arrow). D, Intracellular S aureus reservoirs (pink) colocalized with DAPI-stained host nuclei (gray). E, Immunohistochemistry staining with mouse anti–S aureus (brown) in an NP biopsy section demonstrating S aureus within the cytoplasm of host cells (black arrows) (×40 magnification). DAPI, 4′-6-Diamidino-2-phenylindole, dihydrochloride; FISH, fluorescence in situ hybridization. Journal of Allergy and Clinical Immunology 2015 135, 1648-1651.e5DOI: (10.1016/j.jaci.2014.12.1929) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Immunohistochemical colocalization. Photomicrographs of sequential 2-μm sections of NP tissue stained with anti-AA1 MC tryptase (A) and mouse anti–S aureus (B) demonstrating subepithelial intracellular S aureus within MCs (arrows) (×20 magnification). Both images are shown at higher magnification (C and D ×40 magnification). Journal of Allergy and Clinical Immunology 2015 135, 1648-1651.e5DOI: (10.1016/j.jaci.2014.12.1929) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions