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. 2020 Jul 22;58:102913. doi: 10.1016/j.ebiom.2020.102913

Fig. 3.

Fig. 3

Butyrate suppresses the GC response and autoantibody production in CIA mice

(a, b) CII-specific IgG responses in CIA mice fed the control HAMS or HAMSB diet. The number of CII-specific IgG antibody-forming cells (AFC) cells per 5 × 105 DLN cells three weeks after booster immunization (a, each dot indicates data from 1 well, n= 36 from 6 mice). Statistical analysis was performed on the mean of each mouse. Serum levels of CII-specific IgG1 and IgG2a upon or three weeks after booster immunization (b, n= 5–10).

(c–e) Colonic patches (CoPs) and solitary intestinal lymphoid tissues (SILTs) of CIA mice fed the control HAMS or HAMSB diet. Representative whole-mount B220 immunostaining (brown) of colon tissues (c), and the number of CoPs (d) and SILTs (e) in unimmunized DBA1/J mice and CIA mice one week after the initial immunization (n= 5 or 6).Dashed line, CoP, arrowheads, SILT. Scale bars, 5 mm.

(f–h) GCB cells in CoPs from CIA mice fed the control HAMS or HAMSB diet. Representative flow cytometry contour plots of GL7 and intracellular Bcl-6 staining within CD19+ gate (f), and the frequency (g) and total number (h) of Bcl-6+GL7+ GCB cellstwo weeks after the initial immunization (n= 12).

(i) GC area (mm2) of DLNs from CIA mice fed the control HAMS, or HAMSB diet Data were obtained two weeks after the initial immunization. Each dot indicates a single GC area of DLNs on cross-sections (n= 43, 30). Statistical analysis was performed on the mean of each mouse.

(j–l) GCB cells in DLNs from CIA mice fed control HAMS or HAMSB diet. Representative flow cytometry contour plots of GL7 and intracellular Bcl-6 staining within CD19+ gate (j), and the frequency (k) and total number (l) of Bcl-6+GL7+ GCB cellstwo weeks after the initial immunization (n= 12).

(m) Arthritis scores after antibody cocktail injection of collagen antibody-induced arthritis (CAIA) mice fed control HAMS or HAMSB diet (n= 3; means ± s.e.m.). AUC was calculated for each mice.

Results show one representative experiment of at least two experiments. *P< 0.05, **P< 0.01, ***P< 0.001 (a, b, g–i, k, l, Welch's t-test or unpaired two-tailed Student's t-test; d, e, one-way ANOVA followed by Tukey's posthoc test; m, Log-rank test).