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. 2020 Jun 20;23(7):101288. doi: 10.1016/j.isci.2020.101288

Figure 1.

Figure 1

3O-C12 Stimulates OVA-Specific IgE and IgG1 Production and Enhances Allergic Lung Inflammation

(A) Graphical representation of mouse immunization.

(B) Wild-type C57BL/6 mice (6–8 weeks, male and female) were subcutaneously immunized with OVA (50 μg/mouse) + LPS (0.5 μg/mouse) with or without 3O-C12 (250 μg/mouse) every week for 3 weeks; incomplete Freund adjuvant was used as a vehicle. Mouse serum was collected; OVA-specific IgE, IgG1, IgG2a, IgG2c, IgM, IgA, and IgG2b were analyzed with ELISA. Results are representative of three independent experiments.

(C–E) (C) BALB/c mice were divided into three groups: control (n = 4), asthma (n = 15), and asthma + 3O-C12 (n = 6). The mice were intraperitoneally (i.p.) injected on days 1 and 13 with OVA (50 μg) and aluminum hydroxide (2 mg). On days 20 and 22, the mice were intranasally (i.n.) challenged with OVA (40 μg) and i.p. administered with or without 3O-C12 (500 μg). On days 25, 27, and 29, the mice were i.n. challenged with OVA (40 μg) and i.n. administered with or without 3O-C12 (50 μg). One day after the last OVA challenge, mice were sacrificed and assessed for lung histology (scale bar, 50 μm) (C) and bronchoalveolar lavage fluid (BALF) total cell number (D) and eosinophil number (E).

(F) BMDCs from C57BL/6 mice were stimulated by OVA (200 μg/mL) and LPS (100 ng/mL) overnight, with or without 3O-C12 (10 μM). BMDCs were transferred i.n. to recipients (C57BL/6 mice) (n = 3 in control group; n = 10 in OVA + LPS + DMSO group; n = 11 in OVA + LPS + 3O-C12 group) on days 1 and 12. Each recipient was challenged with 40 μg OVA i.n. on days 13, 14, and 15. One day after the last OVA challenge, mice were sacrificed and assessed for lung histology (scale bar, 50 μm). The statistical quantification (inflammatory cells and airway thickening) of Figures 1C and 1F was analyzed by ImageJ software.

Data are presented as mean ± SD; p values were calculated using the two-tailed Student's t test. Data that did not exhibit a normal distribution were analyzed using the nonparametric Kruskal-Wallis test with Dunn's post hoc test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001; n.s., not significant.