Src homology 2 domain–containing inositol 5-phosphatase 1 deficiency leads to a spontaneous allergic inflammation in the murine lung Sun-Young Oh, PhD,

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Src homology 2 domain–containing inositol 5-phosphatase 1 deficiency leads to a spontaneous allergic inflammation in the murine lung Sun-Young Oh, PhD, Tao Zheng, MD, Monica L. Bailey, BS, Dwayne L. Barber, PhD, John T. Schroeder, PhD, Yoon-Keun Kim, MD, PhD, Zhou Zhu, MD, PhD Journal of Allergy and Clinical Immunology Volume 119, Issue 1, Pages 123-131 (January 2007) DOI: 10.1016/j.jaci.2006.08.029 Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Pulmonary inflammation of SHIP-1−/− mice. Hematoxylin and eosin (H&E) staining of lung sections from WT and SHIP-1−/− mice is shown. Lung sections from WT mice showed normal structure of airways and alveoli. In contrast, mild, medium, and severe cellular infiltration was seen in the airways and lung parenchyma of SHIP-1−/− mice. Journal of Allergy and Clinical Immunology 2007 119, 123-131DOI: (10.1016/j.jaci.2006.08.029) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Pulmonary eosinophilia in SHIP-1−/− mice. A, High-power view of a hematoxylin and eosin (H&E)–stained lung section from a SHIP-1−/− mouse. Enlarged and activated macrophages (arrowheads) and eosinophils forming clusters (arrows) are indicated. B, BAL cellularity analysis. Total cell counts and differentials were obtained from WT mice (open column) and SHIP-1−/− mice (solid column). Each group had 6 mice. ∗P < .01. Journal of Allergy and Clinical Immunology 2007 119, 123-131DOI: (10.1016/j.jaci.2006.08.029) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Mucus metaplasia and airway epithelial hypertrophy. A, PAS staining of lung sections from a WT mouse and a SHIP-1−/− mouse is shown. PAS-positive cells in the airways and thickened airway epithelium were readily seen in SHIP-1−/− mice. The bars show the thickness of the airway epithelium. B, RT-PCR analysis of MUC5AC gene expression in WT and SHIP-1−/− mice is shown. Specific primers were used to amplify MUC5AC mRNA. Shown are representative results of 3 experiments. Journal of Allergy and Clinical Immunology 2007 119, 123-131DOI: (10.1016/j.jaci.2006.08.029) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Subepithelial fibrosis and TGF-β1 expression in SHIP-1−/− mice. A, Masson's trichrome staining of lung sections from a WT mouse and a SHIP-1−/− mouse is shown. The bars show the thickness of the airway epithelium. B, TGF-β1 protein levels in the BAL fluid of WT (open column) and SHIP-1−/−(solid column) mice are shown. ELISA was used to determine the total and spontaneously activated TGF-β1 levels in the BAL fluid and expressed as means ± SEM (n = 6 for each group, ∗P < .05). Journal of Allergy and Clinical Immunology 2007 119, 123-131DOI: (10.1016/j.jaci.2006.08.029) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 Mast cells and histamine content in the lung. A, Toluidine blue staining of lung sections from WT and SHIP-1−/− mice is shown. Degranulating mast cells were readily seen in the lungs of SHIP-1−/− mice. B, Quantification of mast cells is shown. Mast cells were counted and expressed as mean ± SEM numbers of mast cells per view field (n = 4 for each group). Journal of Allergy and Clinical Immunology 2007 119, 123-131DOI: (10.1016/j.jaci.2006.08.029) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 6 Cytokine expression in the lung. A, RT-PCR analysis of total lung RNA from WT and SHIP-1−/− mice using cytokine gene-specific primers is shown. Amplified products were analyzed by means of electrophoresis. Shown is a representative of 3 experiments. B, ELISA analysis of cytokines in the BAL fluid of WT and SHIP-1−/− mice is shown (n = 6-7 mice for each group, ∗P = .01 and ∗∗P < .01 vs WT group). Journal of Allergy and Clinical Immunology 2007 119, 123-131DOI: (10.1016/j.jaci.2006.08.029) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 7 Chemokine expression in the lung. A, RT-PCR analysis of total lung RNA from WT and SHIP-1−/− mice using chemokine gene-specific primers is shown. Amplified products were analyzed by means of electrophoresis. Shown is a representative of 3 experiments. MIP, macrophage inflammatory protein. B, Chemokine concentrations in the BAL fluid of WT and SHIP-1−/− mice were determined by means of ELISA (n = 6-7 mice for each group, ∗P < .05 vs WT group). Journal of Allergy and Clinical Immunology 2007 119, 123-131DOI: (10.1016/j.jaci.2006.08.029) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions