Impact of fucosyltransferase 1-mediated epidermal blood group antigen H on anti-inflammatory response in atopic dermatitis

Abstract

The presence of the blood group H2 antigen on the membrane of red blood cells determines blood type O in individuals and this H2 antigen serves as a precursor to the A and B antigens expressed in blood types A and B, respectively. However, the specific involvement of ABH antigens in skin diseases is unknown. Therefore, we aim to investigate the expression of ABH antigens in skin tissue of patients with atopic dermatitis (AD) and MC903-induced AD-like mice. We demonstrated that the expression of ABH antigen is primarily located in the granular and horny layers of the skin in healthy control individuals. However, in patients with AD, the expression of the ABH antigen was absent or diminished in these layers, while the H2 antigen expression increased in the spinous layers of the affected skin lesions. Then, we investigated the biological function of blood group H antigen mediated by fucosyltransferase 1 (Fut1) in the skin, utilizing an AD mouse model induced by MC903 in wild-type (WT) and Fut1-knockout mice. After the application of MC903, Fut1-deficient mice, with no H2 antigen expression on their skin, exhibited more severe clinical signs, increased ear swelling, and elevated serum IgE levels compared with those of WT mice. Additionally, the MC903-induced thickening of both the epidermis and dermis was more pronounced in Fut1-deficient mice than that in WT mice. Furthermore, Fut1-deficient mice showed a significantly higher production of interleukin-4 (IL-4) and IL-6 in skin lesions compared with that of their WT counterparts. The expression of chemokines, particularly Ccl2 and Ccl8, was notably higher in Fut1-deficient mice compared with those of WT mice. The infiltration of CD4⁺ T cells, eosinophils, and mast cells into the lesional skin was significantly elevated in Fut1-deficient mice compared with that in WT mice. These findings demonstrate the protective role of H2 antigen expression against AD-like inflammation and highlight its potential therapeutic impact on AD through the regulation of blood group antigens.

Keywords: ABH antigens; atopic dermatitis; chemokines; cytokines; fucosyltransferase 1.

Figure 1

The granular layers of healthy skin exhibit ABH antigen and FUT1 expression which is diminished in nonlesional and lesional AD skin. (A) Immunohistochemical staining to detect A and H2 antigens in healthy skin, as well as on nonlesion and lesioned AD skin from individuals with blood type A. (B) Evaluating the expression of A, B, and H2 antigens in the granular (the left graph) and spinous layers (the right graph) of healthy skin, and comparing these expressions in nonlesion and lesional AD skin among individuals with blood types A, B, O, or AB. (C) Immunofluorescent labeling of FUT1 (green) and DAPI (blue) in healthy skin and lesioned AD skin. The data are representative of the mean ± SEM from 39 patients and 66 healthy controls. † < 0.05, †† p < 0.01, *** p < 0.001. AD, atopic dermatitis.

Figure 2

The deficiency of FUT1 exacerbates AD-like inflammation induced by MC903 in a murine model. (A) Immunofluorescent labeling of H2 antigen (green) and DAPI (blue) in the ears of both WT and Fut1-KO mice treated with either vehicle or MC903 on day 12. Scale bars, 20 μm. (B) Immunofluorescent labeling of FUT1 (green) and DAPI (blue) in ear sections from both WT and Fut1-KO mice. Scale bars, 20 μm. (C) Ear skin inflammation induced by MC903 in both WT and Fut1-KO mice on day 12. (D) Clinical scores of MC903-induced ear skin inflammation in both WT and Fut1-KO mice on days 6 and 12. (E) Measurement of ear swelling in both WT and Fut1-KO mice treated with MC903 using a digital caliper. (F) ELISA analysis of serum IgE levels in both WT and Fut1-KO mice treated with either vehicle or MC903 on day 12. (G–I) Immunofluorescent labeling of Loricrin, Filaggrin, or Keratin 10 (green), and DAPI (blue) in ear sections on day 12. The data are representative of the mean ± SEM of three independent experiments, with five mice per group. ns., not significant; * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001. AD, atopic dermatitis; WT, wild-type.

Figure 3

FUT1 deficiency amplifies MC903-induced ear thickening and immune cell infiltration. (A) H&E staining of the ear in both WT and Fut1-KO mice treated with either vehicle or MC903 on day 12. (B) Epidermal thicknesses measurements derived from H&E-stained ear sections. (C) Dermal thicknesses measurements obtained from H&E-stained ear sections. (D) Immunofluorescent labeling of CD4 (green) and DAPI (blue) in ear sections on day 12. (E) Quantitation of CD4⁺ T cells in immunofluorescent-labeled ear sections. (F) Immunofluorescent labeling of Eosinophil (green) and DAPI (blue) in ear sections on day 12. (G) Quantitation of eosinophils in immunofluorescent-labeled ear sections. (H) Toluidine blue staining of mast cells in ear sections. (I) Quantitation of mast cells in toluidine blue-stained ear sections. The data are representative of the mean ± SEM of three independent experiments, with five mice per group. ** p < 0.01, *** p < 0.001 and **** p < 0.0001. AD, atopic dermatitis; H&E, hematoxylin and eosin; WT, wild-type.

Figure 4

FUT1 deficiency enhances the production of AD-associated cytokines in ear skin induced by MC903. (A–D) ELISA analysis of IL-4 (A), IL-5 (B), IL-6 (C), and TSLP (D) levels in the ear skin of both WT and Fut1-KO mice treated with either vehicle or MC903 on day 12. (E, F) Relative mRNA expression of Ccl2 (E) and Ccl8 (F) in the ear skin of both WT and Fut1-KO mice treated with vehicle or MC903 on day 12. The data are representative of the mean ± SEM of three independent experiments, with five mice per group. ns., not significant; * p < 0.05, *** p < 0.001 and **** p < 0.0001. AD, atopic dermatitis; TSLP, thymic stromal lymphopoietin.

Figure 5

FUT1 deficiency heightens the number of IL-4-producing CD4⁺ T cells in the ear-draining LN following MC903 treatment. (A) FACS analysis of IL-4-producing CD4⁺ T cells in in the ear-draining LN after 12 d of MC903 treatment. (B) Quantification of IL-4-producing CD4⁺ T cells in the ear-draining LN on day 12. The data are representative of the mean ± SEM of two independent experiments, with five mice per group. * p < 0.05. LN, lymph node.