TNF can contribute to multiple features of ovalbumin-induced allergic inflammation of the airways in mice  Susumu Nakae, PhD, Carolina Lunderius, PhD,

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TNF can contribute to multiple features of ovalbumin-induced allergic inflammation of the airways in mice  Susumu Nakae, PhD, Carolina Lunderius, PhD, Lien H. Ho, PhD, Beatrix Schäfer, PhD, Mindy Tsai, DMSc, Stephen J. Galli, MD  Journal of Allergy and Clinical Immunology  Volume 119, Issue 3, Pages 680-686 (March 2007) DOI: 10.1016/j.jaci.2006.11.701 Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Reduced expression of antigen induced-AHR after OVA immunization without alum in C57BL/6J-TNF−/− mice. Airway responses to methacholine were measured 24 hours after the last intranasal OVA or PBS challenge. ∗P < .01 for PBS-challenged vs OVA-challenged mice; †P < .01 for OVA-challenged TNF−/− vs wild-type mice. Data were pooled from at least 2 independent experiments, each of which gave similar results. PBS-treated mice, n= 5-8/group; OVA-challenged mice, n = 12-16/group. B, Baseline. Journal of Allergy and Clinical Immunology 2007 119, 680-686DOI: (10.1016/j.jaci.2006.11.701) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Histologic analysis of lungs from C57BL/6J-TNF−/− vs wild-type mice. Lung sections (hematoxylin and eosin–stained) obtained 24 hours after the last intranasal OVA or PBS challenge in mice immunized with OVA without alum. Striking infiltrates of inflammatory cells (∗) are present near the airways in OVA-sensitized, OVA-challenged mice (C-F) but not in PBS-challenged mice (A and B). A-D, ×100, scale bars = 200 μm; E and F, ×400, scale bars = 50 μm. Journal of Allergy and Clinical Immunology 2007 119, 680-686DOI: (10.1016/j.jaci.2006.11.701) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Reduced numbers of leukocytes in BALF; reduced lung levels of MPO, EPO, IL-4, IL-5, and IL-17; and reduced serum levels of OVA-specific IgG1 and IgE in C57BL/6J-TNF−/− mice 24 hours after OVA challenge in mice immunized with OVA without alum. A, Cell numbers in BALFs from the same mice shown in Fig 1. B, MPO and EPO levels in lung homogenates. C, IL-4, IL-5, IL-17, and IFN-γ levels in lung homogenates. D, OVA-specific IgG1 and IgE levels in sera. ∗P < .01 for comparisons shown in brackets; †P < .05 vs values for the corresponding PBS-treated mice. PBS-treated mice, n = 5-8/group; OVA-challenged mice, n = 12-16/group. B, Baseline. Journal of Allergy and Clinical Immunology 2007 119, 680-686DOI: (10.1016/j.jaci.2006.11.701) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Airway responses to methacholine and BALF leukocytes 24 hours after the last intranasal OVA or PBS challenge in C57BL/6J wild-type, TNFR1−/−, TNFR2−/−, and memTNFΔ/Δ mice immunized with OVA without alum. A, Airway responses to methacholine. ∗P < .01 for PBS-treated vs the corresponding OVA-challenged mice; †P < .05 for OVA-treated wild-type vs TNFR1−/− , TNFR2−/−, or memTNFΔ/Δ mice. Data are pooled from the 3 independent experiments that we performed, each of which gave similar results. B, Baseline. B, Cell number in BALFs collected from the same mice shown in A. ∗P < .01 for PBS-treated vs the corresponding OVA-challenged mice; †P < .05 for OVA-treated wild-type vs TNFR1−/−, TNFR2−/−, or memTNFΔ/Δ mice. PBS-treated mice, n = 8-10/group; OVA-treated mice, n = 10-16/group. Journal of Allergy and Clinical Immunology 2007 119, 680-686DOI: (10.1016/j.jaci.2006.11.701) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions