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Critical role of IgE-dependent mast cell activation in a murine model of allergic conjunctivitis  Ken Fukuda, MD, PhD, Masaharu Ohbayashi, PhD, Kei Morohoshi,

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Presentation on theme: "Critical role of IgE-dependent mast cell activation in a murine model of allergic conjunctivitis  Ken Fukuda, MD, PhD, Masaharu Ohbayashi, PhD, Kei Morohoshi,"— Presentation transcript:

1 Critical role of IgE-dependent mast cell activation in a murine model of allergic conjunctivitis 
Ken Fukuda, MD, PhD, Masaharu Ohbayashi, PhD, Kei Morohoshi, MD, PhD, Lane Zhang, BS, Fu-Tong Liu, MD, PhD, Santa J. Ono, PhD  Journal of Allergy and Clinical Immunology  Volume 124, Issue 4, Pages e2 (October 2009) DOI: /j.jaci Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Representative toluidine blue–stained sections of eyes from C57BL/6-Kit+/+ (A), KitW-sh/W-sh (B), and mast cell–reconstituted KitW-sh/W-sh mice that had been subconjunctivally (C) or intravenously (D) injected with BMCMCs. Arrows, Mast cells in conjunctiva; arrowheads, mast cells in eyelid dermis. Scale bar = 100 μm. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Mast cell numbers in conjunctiva and dermis of eyelids of C57BL/6-Kit+/+, KitW-sh/W-sh, and Kit+/+ BMCMC–engrafted KitW-sh/W-sh mice. Dots represent the mast cell numbers from each mouse, and horizontal bars represent the overall mean values from all mice. ND, Not detected; NS, not significant. ∗P < .001 (Scheffe test) versus indicated corresponding values. s.cj., Subconjunctival; i.v., intravenous. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Characterization of conjunctival mast cells in Kit+/+ BMCMC–engrafted KitW-sh/W-sh mice. Reconstituted mast cells in the conjunctiva and dermis of the eyelid were positively stained with the antibodies for mMCP-4 (C), mMCP-5 (D), and mMCP-6 (E) but not with mMCP-2 (B). A, Toluidine blue staining. F, conjunctiva stained with normal rabbit IgG for negative control. Scale bar = 50 μm. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Expression of mRNAs for various mMCPs in conjunctiva of C57BL/6-Kit+/+, KitW-sh/W-sh, and Kit+/+ BMCMC–engrafted KitW-sh/W-sh mice. Amounts of mRNA in the conjunctivas (n = 5 per group) were determined by means of reverse transcription and real-time PCR analysis. Levels were normalized to those of glyceraldehyde-3-phosphate dehydrogenase mRNA and are expressed as mean arbitrary units ± SEMs. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 IgE-mediated early-phase clinical symptoms and mast cell degranulation in C57BL/6-Kit+/+ (A and B), KitW-sh/W-sh (C), and Kit+/+ BMCMC–reconstituted KitW-sh/W-sh mice (D). Clinical appearances (Fig 5, A-D) and scores (E, average ± SE) of early-phase clinical responses are shown. F, The ratio of degranulated mast cells in conjunctiva with passive sensitization by TNP- or RW-specific IgE followed by PBS (open bars) or RW (solid bars) challenge. ND, Not detected. ∗P < .05 versus corresponding values for C57BL/6-Kit+/+ mice sensitized with TNP-IgE. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 6 IgE-mediated late-phase inflammation in C57BL/6-Kit+/+, KitW-sh/W-sh, and Kit+/+ BMCMC–reconstituted KitW-sh/W-sh mice. Mice were passively sensitized by means of transfer of TNP- or RW-specific IgE followed by RW challenge through eye drops. Twenty-four hours after antigen challenge, conjunctivas were harvested for histologic analyses. Infiltrating eosinophils were counted in the lamina propriae. The averages ± SEMs of 5 mice per group are shown. ∗P < .05 versus indicated values. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Representative toluidine blue–stained sections of lungs (A-C), livers (D-F), spleens (G-I), and stomachs (J-L) from C57BL/6-Kit+/+ (Fig E1, A, D, G, and J), KitW-sh/W-sh (Fig E1, B, E, H, and K), and mast cell–reconstituted KitW-sh/W-sh (Fig E1, C, F, I, and L) mice that had been intravenously injected with BMCMCs. Arrows, Mast cells. Scale bar = 100 μm. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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