Figure 1: Cellular loss and WP disorganisation after experimental stroke. | Nature Communications

Figure 1: Cellular loss and WP disorganisation after experimental stroke.

From: Adrenergic-mediated loss of splenic marginal zone B cells contributes to infection susceptibility after stroke

Figure 1

(a) Tiled confocal images of splenic B (red, B220) and T-cell (green, CD3) immunolabelling show a reduction in WP area and disruption to remaining cellular localization. (b) Immunofluorescent labelling of splenic B (red, B220) and T cells (green, CD3) in naïve mice, mice recovered 1–7 days after MCAO or sham-operated mice. (c) Quantification of the percentage area of B220+ immunolabelling per 846.28 μm2 image in spleens from naïve mice or mice recovered from sham or MCAO surgery confirm the area of B-cell labelling is significantly reduced 1–7 days after MCAO in comparison to sham-operated controls. (d) Area of CD3+ T-cell labelling is also reduced. (g) Immunolabelling of CD21/35+ MZ B cells and FDC (red) and F4/80+ macrophages (green) in spleens from naïve mice, mice recovered 1–7 days after MCAO or sham-operated mice show increased macrophages and a reduced area of FDC networks 1–7 days after MCAO in comparison to sham-operated controls. (e) Quantification of CD21/35+ FDC and B-cell immunolabelling confirm a reduced area of labelling 1–7 days after MCAO whereas (f) there is an increase in the area of F4/80+ macrophage labelling (naïve n=4, sham n=4, 1d n=4, 2d n=9, 5d n=5, 7d n=7). (h) Spleen weight, corrected for total body weight, inversely correlates with the infarct volume (n=9). (i) Immunolabelling of B cells (red, B220+) in spleens recovered for 2 days from occlusions of 20 min, 40 min, 60 min or sham surgery demonstrate the extent of disruption to splenic lymphoid tissue is dependent on the extent of injury in the brain. WP area and B-cell follicular structure is preserved in mice after sham surgery or 20 min MCAO. However 40 or 60 min MCAO results in loss of WP and rings of B cells forming within the WP. (Sham n=4, 20 min n=7, 40 min n=7, 60 min n=7). Scale bars (a,b) 200 μm; (g) 100 μm; (i) 500 μm. Data show mean+s.d.; *P<0.05; **P<0.01; ***P<0.001; (c–f) one-way ANOVA with Bonferroni correction comparing all MCAO recovery time points to sham.

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