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. 2019 Aug 5;129(9):3702–3716. doi: 10.1172/JCI93820

Figure 4. CBL stimulates IL-10–producing colonic LP B cells ex vivo.

Figure 4

Unfractionated colonic LP cells from GF or SPF-reared Il10+/EGFP reporter mice were cultured with 0–100 μg/mL CBL from SPF mice for 2 days, as indicated. (A) The frequency of GFP+ cells was assessed by flow cytometry. conc., concentration. (B) Representative dot plots for GFP+ cells in GF colonic cells stimulated with 0 (CBL–), 10, 50, or 100 μg/mL CBL. (C) Using flow cytometry, we determined frequencies of GFP-expressing CD4+ T cells (CD4+CD3+) or B cells (CD19+B220+) as a percentage of total T or B cells using control WT (GFP–) cells that stained with the same antibodies as target samples. (D) Supernatant levels of IL-10 and IL-12p40 were measured by ELISA, and IL-10/IL12 p40 ratios were calculated. Data are presented as median of 8 separated cell cultures, with cells in each culture pooled from 2–4 mice, combined from 3 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, Kruskal-Wallis test with Dunn’s post hoc test.