Murine model of atopic dermatitis associated with food hypersensitivity Xui-Min Li, MDa, Gary Kleiner, MD, PhDa, Chin-Kang Huang, MSa, Soo Yung Lee, MDa,

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Murine model of atopic dermatitis associated with food hypersensitivity Xui-Min Li, MDa, Gary Kleiner, MD, PhDa, Chin-Kang Huang, MSa, Soo Yung Lee, MDa, Brian Schofield, b, Nicholas A. Soter, MDc, Hugh A. Sampson, MDa Journal of Allergy and Clinical Immunology Volume 107, Issue 4, Pages 693-702 (April 2001) DOI: 10.1067/mai.2001.114110 Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 1 Expression of AD-like skin lesions. A, Extensive AD-like skin lesion with raised erythematous eruption, areas of hypopigmentation, and alopecia. B, AD-like lesion about the head and neck with alopecia and excoriations. C, Normal C3H/HeJ mouse. D, Close-up view of erythematous maculopapular eruption with areas of hypopigmentation and alopecia. E, Close-up view of normal skin from shaved naive mouse. Journal of Allergy and Clinical Immunology 2001 107, 693-702DOI: (10.1067/mai.2001.114110) Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 2 AD-like skin lesions before and after therapy with topical corticosteroids. A, Dermatitis and alopecia limited to the facial area. B, Dermatitis and alopecia limited to the facial area before topical corticosteroid therapy. C, Same mouse depicted in B after 2 weeks of topical corticosteroid therapy. No scratching behavior was noted after a few days of therapy, and facial hair had almost completely grown back. D, Extensive AD-like involvement with patches of erythema and hypopigmentation, dry scaly patches, lichenification, and alopecia. E, Same mouse depicted in D after 2 weeks of topical corticosteroid therapy. The mouse had partial hair regrowth and markedly reduced scratching behavior. Nevertheless, the skin retained patchy areas of dry scaly skin and hypopigmentation. Journal of Allergy and Clinical Immunology 2001 107, 693-702DOI: (10.1067/mai.2001.114110) Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 3 Histologic features of AD-like lesions. A, Normal inguinal skin from naive mouse (bar = 100 μm). B, Biopsy specimens from inguinal area of AD-like lesion demonstrating mild spongiosis and epidermal thickening and marked mononuclear round cell infiltrate, especially about the hair follicles (bar = 50 μm). C, Higher magnification of lesional skin depicted in B , showing spongiosis and epidermal thickening. Insert shows eosinophil (bar = 50 μm). D, Frozen section of normal facial skin showing normal number of toluidine blue–stained mast cells (bar = 100 μm). E, Frozen section of lesional facial skin showing markedly increased numbers of toluidine blue–stained mast cells (bar = 100 μm). F, Degranulating mast cells in the dermis of lesional facial skin (bar = 10 μm). A , B , and C are stained with hematoxylin and eosin stain, and D , E , and F are stained with toluidine blue. Journal of Allergy and Clinical Immunology 2001 107, 693-702DOI: (10.1067/mai.2001.114110) Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 4 Increases in serum antigen-specific IgE. Blood was obtained from mice with AD-like lesions and naive mice (n = 4-6) at week 9 and week 20. CM-specific IgE levels were measured with an ELISA. CM-specific IgE levels were highest at week 9 and remained relatively constant between week 9 and week 20. ***P < .001 and *P < .05 versus naive mice. Journal of Allergy and Clinical Immunology 2001 107, 693-702DOI: (10.1067/mai.2001.114110) Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 5 Increased numbers of IgE+ cells in AD-like lesions. A, Frozen section of facial lesion stained with anti-IgE mAb and counter stained with toluidine blue showing dark brown diaminobenzidine-stained IgE+ dendritic-shaped cells in the upper dermis, and IgE+ mast cells in the dermis and muscle (bar = 100 μm). B, IgE+ mast cells with dark brown diaminobenzidine-stained cell membranes and toluidine blue–stained mast cell granules in a facial lesion at higher magnification (bar = 10 μm). C, Frozen section of normal facial skin stained in the same manner as A . Journal of Allergy and Clinical Immunology 2001 107, 693-702DOI: (10.1067/mai.2001.114110) Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 6 Increased numbers of CD4+ T cells in AD-like lesions. A and B are frozen section of the same lesion shown in Fig 5. A, Numerous dark brown diaminobenzidine-stained CD4+ cells, some of which have infiltrated the perifollicular area (bar = 50 μm). B, Few dark brown diaminobenzidine-stained CD8+ cells, most of which are near hair follicles (bar = 50 μm). C and D, Normal facial skin stained in the same manner as in A and B , showing the absence of CD4+ T cells in C and CD8+ T cells in D (bar = 50 μm). Journal of Allergy and Clinical Immunology 2001 107, 693-702DOI: (10.1067/mai.2001.114110) Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 7 Increased IL-5 and IL-13 mRNA expression in AD lesions. Total RNA was extracted from biopsy specimens of lesional skin of mice with AD-like lesions and normal skin of naive mice (n = 4). Semiquantitative RT-PCR was performed in triplicate, as described in the “Methods” section. A, Gel illustrating cytokine (IL-4, IL-5, IL-13, and IFN-γ) mRNA from lesional skin of mice with AD or normal skin from naive mice. β-Actin mRNA expression is shown for comparison. B, OD ratios of cytokine (IL-4, IL-5, IL-13, and IFN-γ) versus β-actin. The results are expressed as means ± SEM of OD ratios of cytokine (IL-4, IL-5, IL-13, and IFN-γ) versus β-actin. ***P < .001 versus naive. Journal of Allergy and Clinical Immunology 2001 107, 693-702DOI: (10.1067/mai.2001.114110) Copyright © 2001 Mosby, Inc. Terms and Conditions