Figure 7.
Manipulating the effects of marginated B cells restrains excessive neutrophilic inflammation in a model of systemic inflammation induced lung injury. (A and B) Lung intravital microscopy quantified neutrophil inflammation and cluster formation in control versus CXCL13 i.t. treated C57BL/6 mice before and after i.v. administration of zymosan-coated beads. Neutrophils (i.v. fluorescently conjugated anti-Ly6G mAb) and neutrophil clusters have been falsely colored white. Scale bars, 200 µm. (C and D) Neutrophil cluster formation and surface area were assessed using lung intravital microscopy in B cell–deficient mice (Ighm−/−) treated with or without exogenous CXCL13 i.t. before and after zymosan-coated bead administration. For A–D, n = 3 independent experiments using 12 mice in total; pooled FOV replicates are shown. Exact P values were calculated using one-way ANOVA with Tukey’s post hoc test. Scale bars, 200 µm. (E) Isolated B cells were cultured, and supernatants were tested for LXA4 levels using ELISA. Adjusted levels were determined by subtracting the control wells without B cells added. n = 3 independent experiments using nine mice total. Exact P values were determined using Student’s t test. (F and G) ELISA quantified LXA4 levels in lung homogenates for C57BL/6 and Ighm−/− mice (F) and mice treated with CXCL13 (G). n = 4 independent experiments using eight mice total. Exact P values were determined using Student’s t test. (H and I) Intravital lung microscopy quantified cluster formation in mice receiving zymosan-coated beads i.v. alone or with i.v. LXA4. Neutrophil clusters have been falsely colored white. Scale bar, 200 µm. Pooled FOV replicates are shown for n = 3 independent experiments using six mice total. Exact P values were determined using Student’s t test. (J) ELISA quantified LXA4 levels in lung homogenates for C57BL/6 and Ighm−/− mice 1 h after administration of i.v. zymosan-coated beads. n = 4 independent experiments using eight mice total. Exact P values were determined using Student’s t test. Pooled data are presented as mean ± SD. *, P < 0.05; **, P < 0.01; ****, P < 0.0001.

Manipulating the effects of marginated B cells restrains excessive neutrophilic inflammation in a model of systemic inflammation induced lung injury. (A and B) Lung intravital microscopy quantified neutrophil inflammation and cluster formation in control versus CXCL13 i.t. treated C57BL/6 mice before and after i.v. administration of zymosan-coated beads. Neutrophils (i.v. fluorescently conjugated anti-Ly6G mAb) and neutrophil clusters have been falsely colored white. Scale bars, 200 µm. (C and D) Neutrophil cluster formation and surface area were assessed using lung intravital microscopy in B cell–deficient mice (Ighm−/−) treated with or without exogenous CXCL13 i.t. before and after zymosan-coated bead administration. For A–D, n = 3 independent experiments using 12 mice in total; pooled FOV replicates are shown. Exact P values were calculated using one-way ANOVA with Tukey’s post hoc test. Scale bars, 200 µm. (E) Isolated B cells were cultured, and supernatants were tested for LXA4 levels using ELISA. Adjusted levels were determined by subtracting the control wells without B cells added. n = 3 independent experiments using nine mice total. Exact P values were determined using Student’s t test. (F and G) ELISA quantified LXA4 levels in lung homogenates for C57BL/6 and Ighm−/− mice (F) and mice treated with CXCL13 (G). n = 4 independent experiments using eight mice total. Exact P values were determined using Student’s t test. (H and I) Intravital lung microscopy quantified cluster formation in mice receiving zymosan-coated beads i.v. alone or with i.v. LXA4. Neutrophil clusters have been falsely colored white. Scale bar, 200 µm. Pooled FOV replicates are shown for n = 3 independent experiments using six mice total. Exact P values were determined using Student’s t test. (J) ELISA quantified LXA4 levels in lung homogenates for C57BL/6 and Ighm−/− mice 1 h after administration of i.v. zymosan-coated beads. n = 4 independent experiments using eight mice total. Exact P values were determined using Student’s t test. Pooled data are presented as mean ± SD. *, P < 0.05; **, P < 0.01; ****, P < 0.0001.

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