Figure 6.
ACC1 controls MC903-induced skin inflammation via IL-3–mediated basophil activation. (A) Intracellular staining profiles of IL-3 and ACC1 in stimulated ear-dLN CD44hiCD4+ T cells. Histogram shows ACC1 expression in IL-3+ or IL-3− cells. (B) Intracellular staining profiles of IL-3 in stimulated ear-dLN CD44hiCD4+ T cells. (C) Ear thickness of Acaca+/+ and Acaca−/− mice after daily MC903 treatment (2 nmol per ear). Values are differences in thickness from day 0. (D and E) Representative images of ears collected from Acaca+/+ or Acaca−/− mice topically treated with MC903 (D) and histological analysis of ear sections fixed and stained with H&E (HE; E). Scale bars represent 100 µm. (F) Clinical score of the ear skin based on the redness and swelling. (G) Expression of CD63 on ear-dLN basophils (CD45intCD200R3+CD49b+c-kit−) of Acaca+/+ or Acaca−/− mice. (H and I) Representative images of ears collected from Acaca+/+ and Acaca−/− mice subcutaneously administered with mIL-3/anti-IL-3 or PBS to shoulders (H). Histological analysis of ear sections of mice fixed and stained with H&E (I). Scale bars represent 100 µm. (J) Ear thickness of Acaca+/+ and Acaca−/− mice subcutaneously injected with mIL-3/anti-IL-3 mixture or PBS, after daily MC903 treatment. Values are differences in thickness from day 0. P values indicated comparison with Acaca+/+ group administered PBS and Acaca+/+ or Acaca−/− group administered IL-3 mixture. (K) Clinical score of the ear skin based on redness and swelling. (L) Expression of CD63 on ear-dLN basophils (CD45intCD200R3+CD49b+c-kit−) of Acaca+/+ and Acaca−/− mice subcutaneously injected with mIL-3/anti-IL-3 or PBS, after daily MC903 treatment. (M) Quantitative RT-PCR analysis of Cd203c in tissue section of ear, same condition as in L. For each group, n = 4 (B–E and J–M); n = 6 (A); n = 3–4 (G); or n = 4–5 (F) biologically independent samples are shown. More than three independent experiments were performed with similar results for A–M. Mean values with SD are shown for A–C, F–G, and J–M. Two-way ANOVA was applied for C and J. An unpaired two-tailed Student’s t test was applied for A, B, F, G, and K–M. Statistical significance (P values) is indicated as *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; N.S., not significant.

ACC1 controls MC903-induced skin inflammation via IL-3–mediated basophil activation. (A) Intracellular staining profiles of IL-3 and ACC1 in stimulated ear-dLN CD44hiCD4+ T cells. Histogram shows ACC1 expression in IL-3+ or IL-3− cells. (B) Intracellular staining profiles of IL-3 in stimulated ear-dLN CD44hiCD4+ T cells. (C) Ear thickness of Acaca+/+ and Acaca−/− mice after daily MC903 treatment (2 nmol per ear). Values are differences in thickness from day 0. (D and E) Representative images of ears collected from Acaca+/+ or Acaca−/− mice topically treated with MC903 (D) and histological analysis of ear sections fixed and stained with H&E (HE; E). Scale bars represent 100 µm. (F) Clinical score of the ear skin based on the redness and swelling. (G) Expression of CD63 on ear-dLN basophils (CD45intCD200R3+CD49b+c-kit−) of Acaca+/+ or Acaca−/− mice. (H and I) Representative images of ears collected from Acaca+/+ and Acaca−/− mice subcutaneously administered with mIL-3/anti-IL-3 or PBS to shoulders (H). Histological analysis of ear sections of mice fixed and stained with H&E (I). Scale bars represent 100 µm. (J) Ear thickness of Acaca+/+ and Acaca−/− mice subcutaneously injected with mIL-3/anti-IL-3 mixture or PBS, after daily MC903 treatment. Values are differences in thickness from day 0. P values indicated comparison with Acaca+/+ group administered PBS and Acaca+/+ or Acaca−/− group administered IL-3 mixture. (K) Clinical score of the ear skin based on redness and swelling. (L) Expression of CD63 on ear-dLN basophils (CD45intCD200R3+CD49b+c-kit−) of Acaca+/+ and Acaca−/− mice subcutaneously injected with mIL-3/anti-IL-3 or PBS, after daily MC903 treatment. (M) Quantitative RT-PCR analysis of Cd203c in tissue section of ear, same condition as in L. For each group, n = 4 (B–E and J–M); n = 6 (A); n = 3–4 (G); or n = 4–5 (F) biologically independent samples are shown. More than three independent experiments were performed with similar results for A–M. Mean values with SD are shown for A–C, F–G, and J–M. Two-way ANOVA was applied for C and J. An unpaired two-tailed Student’s t test was applied for A, B, F, G, and K–M. Statistical significance (P values) is indicated as *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; N.S., not significant.

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