Α(1,3) Fucosyltransferases IV and VII Are Essential for the Initial Recruitment of Basophils in Chronic Allergic Inflammation Kazumi Saeki, Takahiro.

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α(1,3) Fucosyltransferases IV and VII Are Essential for the Initial Recruitment of Basophils in Chronic Allergic Inflammation Kazumi Saeki, Takahiro Satoh, Hiroo Yokozeki Journal of Investigative Dermatology Volume 133, Issue 9, Pages 2161-2169 (September 2013) DOI: 10.1038/jid.2013.160 Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 IgE-mediated chronic allergic inflammation (IgE-CAI) in α(1, 3) fucosyltransferase-IV (FT-IV)- and/or FT-VII-deficient mice. (a) IgE-CAI was induced in mice lacking FT-IV and/or FT-VII. Negative control mice were challenged with trinitrophenyl-OVA (TNP-OVA) without TNP-IgE injection. (b) Histopathological features of the skin (Giemsa’s staining). (c) Cell populations in inflammatory skin. (d) Basophil numbers in inflammatory skin. Basophils were detected by mouse mast cell protease-8 mAb (arrows in the right panel). *P<0.05 compared with wild-type (WT) mice. BM, basement membrane. Bar=100μm. Journal of Investigative Dermatology 2013 133, 2161-2169DOI: (10.1038/jid.2013.160) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Delayed-type hypersensitivity (DTH) reactions in α(1, 3) fucosyltransferase-IV (FT-IV)- and/or FT-VII-deficient mice. Contact hypersensitivity (CHS) and DTH to sheep red blood cells (DTH-SRBCs) were induced in mice lacking FT-IV and/or -VII. *P<0.05 compared with wild-type (WT) mice. Journal of Investigative Dermatology 2013 133, 2161-2169DOI: (10.1038/jid.2013.160) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 α(1, 3) Fucosyltransferase-IV/VII (FT-IV/VII) in basophils are indispensable for IgE-mediated chronic allergic inflammation (IgE-CAI). IgE-CAI was induced in FcRγ(-/-) mice that received primary basophils from wild-type (WT), FT-IV(-/-), FT-VII(-/-), and FT-IV(-/-)/FT-VII(-/-) mice. (a) Although basophils from FT-IV(-/-) and FT-VII(-/-) mice induced exacerbated IgE-CAI as compared with WT basophils, (b) FT-IV(-/-)/FT-VII(-/-) mice–derived basophils were incapable of inducing IgE-CAI. *P<0.05. Journal of Investigative Dermatology 2013 133, 2161-2169DOI: (10.1038/jid.2013.160) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Selectin-dependent cooperative recruitment of basophils and effector cells. (a) Irradiated α(1, 3) fucosyltransferase-IV (FT-IV)(-/-)/FT-VII(-/-) mice received wild-type (WT) basophils in combination with CD49b(-) bone marrow cells (effector cells) from either WT or FT-IV(-/-)/FT-VII(-/-) mice. They were then immunized with trinitrophenyl-IgE (TNP-IgE) and challenged with TNP-OVA. The untreated group comprised FT-IV(-/-)/FT-VII(-/-) mice without cell transfer. (b) Cell populations in inflammatory skin. *P<0.05 compared with WT effector cells. BM, basement membrane. Journal of Investigative Dermatology 2013 133, 2161-2169DOI: (10.1038/jid.2013.160) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Expression of α(1, 3) fucosyltransferase (FT) messenger RNA (mRNA) and FT-dependent selectin binding in basophils. (a) Primary basophils were subjected to further purification by positive selection with CD123 (purity >99%). Transcripts for FT-IV and FT-VII mRNA were quantified by real-time PCR. (b) Binding of soluble P- and E-selectins to primary basophils assessed by flow cytometry. BMBa, bone marrow–derived basophil; BMMCs, bone marrow–derived mast cells. Journal of Investigative Dermatology 2013 133, 2161-2169DOI: (10.1038/jid.2013.160) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 6 Basophil P-selectin glycoprotein-1 (PSGL-1)–L-selectin interaction involvement in IgE-mediated chronic allergic inflammation (IgE-CAI). (a) PSGL-1 and L-selectin expressions on basophils. (b) PSGL-1 Ab effect on selectin binding to primary basophils. (c) L-selectin binding to basophils from wild-type (WT) and α(1, 3) fucosyltransferase-IV (FT-IV)(-/-)/FT-VII(-/-) mice. (d) Effects of L-selectin and/or PSGL-1 Abs on IgE-CAI in WT mice. *P<0.05 compared with control IgG. **P<0.05 compared with L-selectin or PSGL-1 Ab alone. Journal of Investigative Dermatology 2013 133, 2161-2169DOI: (10.1038/jid.2013.160) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions