Images in allergy and immunology: Regulatory T cells in allergic disease Susan C. Foley, MD, David Préfontaine, MSc, Michelle D'Antoni, BSc, Qutayba Hamid,

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Images in allergy and immunology: Regulatory T cells in allergic disease Susan C. Foley, MD, David Préfontaine, MSc, Michelle D'Antoni, BSc, Qutayba Hamid, MD, PhD Journal of Allergy and Clinical Immunology Volume 120, Issue 2, Pages 482-486 (August 2007) DOI: 10.1016/j.jaci.2007.06.036 Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Infiltration by T cells in allergic disease. Immunostaining with CD4 antibody showing infiltration by CD4-expressing T cells in the nasal submucosa and epithelium in allergic rhinitis (A) and in the skin in acute atopic dermatitis (B). Magnification ×200. Reprinted with permission from Fiset PO, Leung DYM, Hamid Q. Immunopathology of atopic dermatitis. J Allergy Clin Immunol 2006;118:287-90 (B). Journal of Allergy and Clinical Immunology 2007 120, 482-486DOI: (10.1016/j.jaci.2007.06.036) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Diagram showing control of allergic airways disease by Treg cells. CD4+CD25+ and IL-10–producing Treg cells can regulate allergic sensitization in vivo through inhibitory effects on TH2 cells or on dendritic cells in the lung. The dendritic cells produce immunomodulatory cytokines such as IL-6, which has been shown to inhibit CD4+CD25+ Treg cell function, and IL-10, which has been shown to induce IL-10–producing Treg cells. TCR, T-cell receptor. Reprinted with permission from Hawrylowicz CM. Regulatory T cells and IL-10 in allergic inflammation. J Exp Med 2005;202:1459-63. Journal of Allergy and Clinical Immunology 2007 120, 482-486DOI: (10.1016/j.jaci.2007.06.036) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Disease manifestations of Foxp3 deficiency in human subjects. A, Eczemalike skin lesions on the face of a child with IPEX. B, Enteritis-like picture in IPEX. Biopsy specimen of the sigmoid colon in a child with IPEX showing a colitis-like picture with infiltration by a mixed cellular infiltrate (hematoxylin and eosin staining). Adapted from Chatila TA. Role of regulatory T cells in human diseases. J Allergy Clin Immunol 2005;116:949-59.1 Journal of Allergy and Clinical Immunology 2007 120, 482-486DOI: (10.1016/j.jaci.2007.06.036) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 FOXP3-expressing T lymphocytes detected in PBMCs from children display TLR4 immunfluorescence.8 PBMC cytospins were stained with mouse antihuman FOXP3 Ab (red), biotinylated LPS (green), and Hoechst 33342 nuclear staining (Molecular Probes, Eugene, Ore) (blue). FOXP3-expressing cells have pink nuclei. These cells display TLR4 immunoreactivity at the cell surface. Journal of Allergy and Clinical Immunology 2007 120, 482-486DOI: (10.1016/j.jaci.2007.06.036) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 Immunofluorescent colocalization of TLR4 expression by CD3+ T cells in the nasal mucosa of children. CD3+ T cells appear red (A), TLR4-positive cells appear green (B), and CD3+ T cells that colocalize with TLR4 appear yellow (C). Adapted with permission from Tulic MK, Fiset PO, Manoukian JJ, Frenkiel S, Lavigne F, Eidelman DH, et al. Roll of toll-like receptor 4 in protection by bacterial lipopolysaccharide in the nasal mucosa of atopic children but not adults. Lancet 2004;363:1689-97.9 Journal of Allergy and Clinical Immunology 2007 120, 482-486DOI: (10.1016/j.jaci.2007.06.036) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 6 Immunofluorescence demonstrating IL-10 production (A, red) by TLR4-expressing cells (B, green) in the nasal mucosa of atopic children. C, Colocalization of TLR4 with IL-10 immunoreactivity appears yellow. Adapted with permission from Tulic MK, Fiset PO, Manoukian JJ, Frenkiel S, Lavigne F, Eidelman DH, et al. Roll of toll-like receptor 4 in protection by bacterial lipopolysaccharide in the nasal mucosa of atopic children but not adults. Lancet 2004;363:1689-97.9 Journal of Allergy and Clinical Immunology 2007 120, 482-486DOI: (10.1016/j.jaci.2007.06.036) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 7 Colocalization of CD4+ CD25+ T cells in the nasal mucosa from explanted atopic children. Ex vivo LPS stimulation of mucosal tissue sections applied before specific allergen challenge (A) induced an increase in the numbers of CD4+CD25+ T cells (yellow) compared with allergen alone (B). CD4+ cells appear red, whereas activated CD25-expressing cells are green. Adapted with permission from Tulic MK, Fiset PO, Manoukian JJ, Frenkiel S, Lavigne F, Eidelman DH, et al. Roll of toll-like receptor 4 in protection by bacterial lipopolysaccharide in the nasal mucosa of atopic children but not adults. Lancet 2004;363:1689-97.9 Journal of Allergy and Clinical Immunology 2007 120, 482-486DOI: (10.1016/j.jaci.2007.06.036) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 8 Antiallergic activities of immunostimulatory sequence-oxydeoxynucleotide (ISS-ODN) or CpG motifs. DNA containing 1 or more CpG motifs is taken up by endocytosis in most cell types but only activates cells of the innate arm of the immune system that expresses TLR9 (B cells and plasmacytoid dendritic cells). These cells create a TH1-like cytokine milieu by secreting IFN-α, IFN-β, IL-12, IFN-inducible protein 10 (IP-10) and other TH1-promoting cytokines and chemokines. Natural killer (NK) cells are secondarily activated and secrete IFN-γ. Over time, ISS-ODN–induced innate immune activation imprints on adaptive immunity by promoting the development of TH1-biased and inhibiting TH2-biased adaptive responses to allergens. Reprinted with permission from Horner AA. Toll-like receptor ligands and atopy: a coin with at least 2 sides. J Allergy Clin Immunol 2006;117:1133-40.13 Journal of Allergy and Clinical Immunology 2007 120, 482-486DOI: (10.1016/j.jaci.2007.06.036) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions