Prevention of leukocyte migration to inflamed skin with a novel fluorosugar modifier of cutaneous lymphocyte-associated antigen
Charles J. Dimitroff, … , Thomas S. Kupper, Robert Sackstein
Charles J. Dimitroff, … , Thomas S. Kupper, Robert Sackstein
Published October 1, 2003
Citation Information: J Clin Invest. 2003;112(7):1008-1018. https://doi.org/10.1172/JCI19220.
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Article Dermatology

Prevention of leukocyte migration to inflamed skin with a novel fluorosugar modifier of cutaneous lymphocyte-associated antigen

Abstract

E-selectin and P-selectin on dermal postcapillary venules play critical roles in the migration of effector T cells into inflamed skin. P-selectin glycoprotein ligand-1 (PSGL-1) modified by α1,3-fucosyltransferase is the principal selectin ligand on skin-homing T cells and is required for effector T cell entry into inflamed skin. We have previously shown that a fluorinated analog of N-acetylglucosamine peracetylated-4-fluorinated-D-glucosamine (4-F-GlcNAc), inhibits selectin ligand expression on human T cell PSGL-1. To analyze 4-F-GlcNAc efficacy in dampening effector T cell migration to inflamed skin, we elicited allergic contact hypersensitivity (CHS) reactions in mice treated with 4-F-GlcNAc. We also investigated 4-F-GlcNAc efficacy on lymphocyte E-selectin ligand expression in LNs draining antigen-sensitized skin and on other immunological processes requisite for CHS responses. Our results showed that 4-F-GlcNAc treatment attenuated lymphocyte E-selectin ligand expression in skin-draining LNs and prevented CHS reactions. Significant reductions in inflammatory lymphocytic infiltrate were observed, while pathways related to antigenic processing and presentation and naive T cell recognition within skin-draining LNs were unaffected. These data indicate that 4-F-GlcNAc prevents CHS by inhibiting selectin ligand activity and the capacity of effector T cells to enter antigen-challenged skin without affecting the afferent phase of CHS.

Authors

Charles J. Dimitroff, Thomas S. Kupper, Robert Sackstein

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Effects of 4-F-GlcNAc on effector lymphocyte E-selectin ligand and PSGL-...
Effects of 4-F-GlcNAc on effector lymphocyte E-selectin ligand and PSGL-1 expression. Lymphocytic lysates were prepared from inguinal LNs draining DNFB-sensitized skin after day 7 of DNFB sensitization. Lysates (40 μg/lane) were separated on reducing 4–20% SDS-PAGE gradient gels, transferred to PVDF membrane, and blotted with mouse E-selectin–Ig (1 μg/ml) (a) or rabbit anti-sera against mouse PSGL-1 (0.5 μg/ml) (b) and relevant AP-conjugated secondary Ab’s (1:2,000) and AP substrate solution. Negative control blots, E-selectin–Ig staining in the presence in 5 mM EDTA or AP-conjugated secondary Ab’s alone, did not show any signal. Please note the induction of lymphocyte E-selectin ligand principally by the 120- and 220-kDa isoforms of PSGL-1, as well as by a 190-kDa glycoprotein from skin-draining LNs of DNFB-sensitized mice (a). Also, though PSGL-1 levels were relatively unchanged (b), lymphocyte E-selectin ligand induction from skin-draining LNs of DNFB-sensitized FucTIV/VII–/– mice or of DNFB-sensitized mice treated with 100 mg/kg 4-F-GlcNAc was absent (a).