Molecular pathology of allergic disease

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Presentation transcript:

Molecular pathology of allergic disease Qutayba A. Hamid, MD, PhD, Eleanor M. Minshall, PhD Journal of Allergy and Clinical Immunology Volume 105, Issue 1, Pages 20-36 (January 2000) DOI: 10.1016/S0091-6749(00)90172-6 Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 1 Histologic and molecular pathologic features of bronchial asthma. Low-power view of bronchial biopsy specimen that has undergone in situ hybridization for IL-5 messenger RNA (mRNA) is shown (A) . Note relatively well-preserved histologic features and large number of IL-5 mRNA-positive cells that can be seen as dark aggregations of silver grains. Representative example (B) of bronchial biopsy specimen taken from asthmatic subject demonstrating marked inflammatory cell infiltrate a thickening of lamina reticularis (subepithelial fibrosis). Epithelium in this specimen is well preserved. C, Dark-field illumination of in situ hybridization for IL-5 mRNA in bronchial biopsy specimen from asthmatic subject. IL-5 mRNA-positive cells are observed as collections of silver grains and have been colocalized to CD3+ T lymphocytes. IL-4 mRNA-positive cells are shown as dark aggregations of silver grains in light-field illumination in bronchoalveolar lavage (BAL) cells from patients with asthma (D) . Again, the majority of these cells have been shown to be T lymphocytes. E and F, Fluorescent micrographs of simultaneous in situ hybridization, IL-5 mRNA, and immunocytochemistry for CD3+, respectively, demonstrating that the majority of IL-5 mRNA-positive cells are lymphocytes. Journal of Allergy and Clinical Immunology 2000 105, 20-36DOI: (10.1016/S0091-6749(00)90172-6) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 2 Peripheral blood eosinophils are a potent source of cytokines. The majority of eosinophils from peripheral blood of healthy subjects expresses IL-12 mRNA as detected with use of in situ hybridization and dark-field illumination (A) . IL-12 mRNA-positive signals are seen as silver grains overlying individual cells. Peripheral blood eosinophils from asthmatic subjects also have the capacity to stain positive for IL-6 immunoreactivity (B) . Positive immunoreactivity is visualized as red cytoplasmic staining. Journal of Allergy and Clinical Immunology 2000 105, 20-36DOI: (10.1016/S0091-6749(00)90172-6) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 3 IL-5-immunoreactivity within bone marrow and locally differentiating MBP-positive eosinophils in nasal explants. Immunocytochemistry for IL-5-immunoreactive protein in bone marrow of Balb/c mice 6 hours after antigen challenge (A) . Number of cells expressing Il-5 immunoreactive protein is increased as a result of antigen exposure in previously sensitized animals and is predominantly localized to CD3+ lymphocytes. B, Number of MBP-positive eosinophils within nasal explants 6 hours after allergen challenge. In absence of de novo inflammatory cell recruitment, MBP-positive cells are believed to differentiate from locally residing progenitor cells that express CD34 cell marker. Journal of Allergy and Clinical Immunology 2000 105, 20-36DOI: (10.1016/S0091-6749(00)90172-6) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 4 Confocal microscopy of IL-5R mRNA detected by FITC-labeled probe showing serial sections through eosinophils and 3-dimensional imaging. Expression of IL-5R was up-regulated in both atopic and nonatopic asthmatic subjects. Journal of Allergy and Clinical Immunology 2000 105, 20-36DOI: (10.1016/S0091-6749(00)90172-6) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 5 Expression of IL-9 mRNA and immunoreactive protein in asthma and sarcoidosis, a nonallergic lung disease. Representative examples of in situ hybridization of IL-9 mRNA with use of sulfur 35–labeled complementary RNA probes in bronchial biopsy specimens from asthmatic subjects. (A, Original magnification ×200; B , original magnification ×400). C, Expression of IL-9 mRNA in bronchial biopsy specimen from patient with sarcoidosis. (Original magnification ×400). IL-9 immunoreactivity is shown in atopic asthmatic subject (D) , in patient with sarcoidosis (E) , and in nonatopic nonasthmatic control subject (F) . Journal of Allergy and Clinical Immunology 2000 105, 20-36DOI: (10.1016/S0091-6749(00)90172-6) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 6 Expression of eotaxin mRNA and immunoreactive protein in asthma. Cellular distribution or eotaxin mRNA (A to D ) and immunoreactive protein (E to H ) within the airways (A, B, E, F ) and BAL cells (C, D, G, H ) of asthmatic patients (A) compared with healthy control subjects (B) . Staining was observed exclusively in the airway’s epithelium and in many infiltrating cells in asthmatic biopsy specimens. Representative cytospin preparations of BAL cells obtained from asthmatic patient (C) and nonasthmatic control subject (D) hybridized with FITC-labeled antisense riboprobe to eotaxin. Immunostaining for eotaxin-immunoreactive protein within the airways (E, F ) and BAL cells of asthmatic (G) and nonasthmatic control (H) subjects, respectively. Tissue sections and BAL cells were fixed and stained with rabbit polyclonal antieotaxin antibody. Alkaline phosphatase–conjugated goat antirabbit IgG and fast red substrate or FITC-conjugated swine antirabbit IgG and fast red substrate or FITC-conjugated swine antirabbit IgG were used to visualize eotaxin imunoreactivity. Journal of Allergy and Clinical Immunology 2000 105, 20-36DOI: (10.1016/S0091-6749(00)90172-6) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 7 Expression of TGF-β1 and its association with eosinophils in bronchial biopsy specimens. Immunostaining for EG2-positive cells in mild (a) and severe (b) asthmatic bronchial biopsy specimens and in situ hybridization detecting TGF-β1 mRNA-positive cells in a moderate (c) and a severe (d) asthma. Inset in d, TGF-β1 mRNA-positive cell, indicated by the silver grains, associated with EG2+ immunoreactivity (pink staining). Immunocytochemistry for TGF-β1 immunoreactive protein in moderate asthma (e) and double immunostaining demonstrating presence of TGF-β1 immunoreactivy in EG2+ eosinphils (f) . Hematoxylin and eosin staining of bronchial biopsy specimen from a patient with severe asthmatic (g) and representative examples of subepithelial fibrosis as detected by van Gieson’s staining in mild (h) , moderate (j) , and severe (k) asthma. (Reproduced with permission from Minshall EM, Leung DY, Martin RJ, Song YL, Cameron L, Ernst P, et al. Eosinophil-associated TGF-β mRNA expression and airways fibrosis in bronchial asthma. Am J Respir Cell Mol Biol 1997;17:326-33.) Journal of Allergy and Clinical Immunology 2000 105, 20-36DOI: (10.1016/S0091-6749(00)90172-6) Copyright © 2000 Mosby, Inc. Terms and Conditions