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IL-4– and IL-5–positive T lymphocytes, eosinophils, and mast cells in allergen-induced late-phase cutaneous reactions in atopic subjects  Luis T. Barata,

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Presentation on theme: "IL-4– and IL-5–positive T lymphocytes, eosinophils, and mast cells in allergen-induced late-phase cutaneous reactions in atopic subjects  Luis T. Barata,"— Presentation transcript:

1 IL-4– and IL-5–positive T lymphocytes, eosinophils, and mast cells in allergen-induced late-phase cutaneous reactions in atopic subjects  Luis T. Barata, MDa, Sun Ying, PhDa, Qiu Meng, MDa, Julia Barkansa, Karalasingam Rajakulasingam, MDa, Stephen R. Durham, MDb, A.Barry Kay, MDa  Journal of Allergy and Clinical Immunology  Volume 101, Issue 2, Pages (February 1998) DOI: /S (98) Copyright © 1998 Mosby, Inc. Terms and Conditions

2 Fig. 1 Time course of the allergen-induced cutaneous late-phase response (top panel) and the expression of IL-4 and IL-5 mRNA+ and protein product (middle and bottom panels, respectively). Solid circles represent cytokine mRNA cells detected by in situ hybridization; open circles represent cytokine protein product positive cells detected by immunohistochemistry. Values are expressed as mean ± SEM (n  = 6). Journal of Allergy and Clinical Immunology  , DOI: ( /S (98) ) Copyright © 1998 Mosby, Inc. Terms and Conditions

3 Fig. 2 Total numbers of CD3+ T cells, EG2+ eosinophils, tryptase-positive mast cells and CD68 macrophages (cross-hatched bars), and IL- 4+/phenotype positive double-stained cells (solid bars) in skin biopsy specimens taken 24 hours after allergen challenge in six atopic subjects. Cytokine-positive cells were detected either by in situ hybridization (top panel) or immunohistochemistry (bottom panel). Values are expressed as mean ± SEM (n = 6). There was no significant difference in the numbers of IL-4 mRNA or protein-positive cells with each phenotypic marker. Journal of Allergy and Clinical Immunology  , DOI: ( /S (98) ) Copyright © 1998 Mosby, Inc. Terms and Conditions

4 Fig. 3 Total numbers of CD3+ T cells, EG2+ eosinophils, tryptase-positive mast cells and CD68+ macrophages (cross-hatched bars), and IL- 5+/phenotype+ double-stained cells (solid bars) in skin biopsy specimens taken 24 hours after allergen challenge in six atopic subjects. Cytokine-positive cells were detected either by in situ hybridization (top panel) or by immunohistochemistry (bottom panel). Values are expressed as mean ± SEM (n = 6). There was no significant difference in the numbers of IL-5 mRNA or protein-positive cells with each phenotypic marker. Journal of Allergy and Clinical Immunology  , DOI: ( /S (98) ) Copyright © 1998 Mosby, Inc. Terms and Conditions

5 Fig. 4 Immunohistochemistry and ISH studies of skin biopsies 24 hours after allergen challenge. A, Single ISH for IL-4 mRNA+ cells (arrows); B, double immunohistochemistry/ISH colocalization of IL-4 mRNA to EG2+ eosinophils (arrows); C, single immunohistochemistry shows IL-4 immunoreactive cells with the method of ABC. Positive cells exhibit red color (some positive cells are indicated by arrows). D, High power of C. E, Negative control of immunohistochemistry with irrelevant antibody. F, Single immunohistochemistry of IL-5 immunoreactive cells (arrows). G and H, Examples of double immunohistochemistry showing colocalization of IL-4 and IL-5 immunoreactivity (protein product) to tryptase-positive mast cell (red/brown, arrow), EG2+ eosinophil (red/brown, arrow), and single EG2 eosinophil (brown, arrowhead), respectively. Journal of Allergy and Clinical Immunology  , DOI: ( /S (98) ) Copyright © 1998 Mosby, Inc. Terms and Conditions

6 Fig. 5 RT-PCR detection of IL-4, IL-5, and β-actin mRNA in blood eosinophils (lanes 3-13). M, molecular marker. Lane 1 and 2: positive and negative controls. Lane 3: unstimulated eosinophils. Lane 4-13: eosinophils incubated for 30 minutes and 120 minutes with HBSS-coated particles (lane 4-5), albumin-coated particles (lane 6-7), serum-coated particles (lane 8-9), IgG-coated particles (lane 10-11), and sIgA-coated particles (lane 12-13), respectively. Journal of Allergy and Clinical Immunology  , DOI: ( /S (98) ) Copyright © 1998 Mosby, Inc. Terms and Conditions

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