Figure 1.
Figure 1. Thrombosis in the spleen and liver follows different kinetics after STm infection. (A) Frozen spleens from WT mice infected with 5 × 105 STm intraperitoneally (IP) were sectioned longitudinally to the hilum (≥1200 μm deep), and 5-μm sections were stained by IHC. Scans of stained spleen sections from mice infected for 0, 1, 7, and 21 days show blood vessels identified with anti-CD31 in blue and thrombi with antifibrin/fibrinogen in brown. Arrows identify individual thrombi. (B) Representative low power images of spleen and liver sections stained for CD41 (platelets; blue) and fibrin/fibrinogen (brown) from mice infected for 24 hours via the IV route with 5 × 105 STm SL3261. Arrows identify individual thrombi. The left-hand graph shows the number of thrombi per spleen section for mice infected IP or IV. The right-hand graph shows the number of thrombi per section in the spleen and liver from these IV-infected mice. (C) Representative low-power images of spleen and liver sections stained for CD41 (platelets; blue) and fibrin/fibrinogen (brown) from mice infected for 24 hours with the 105 virulent STm SL1344 strain. Arrows identify individual thrombi. The left-hand graph shows the number of thrombi per spleen section for mice infected with SL3261 (attenuated) or SL1344 (virulent). The right-hand graph shows the number of thrombi per section in the spleen and liver from mice infected with SL1344. (D) Representative scans at higher magnification of spleen sections (top) and liver sections (bottom) from WT mice at days 0, 1, 7, and 21 after infection with 5 × 105 STm SL3261. Sections are stained for fibrin/fibrinogen (brown) and CD41 (blue). RP, red pulp; T, thrombus; V, vein; WP, white pulp. (E) Quantification of numbers of thrombi per spleen section (left graph) and the proportion of section area covered by thrombi (right graph) at days 0, 1, 2, 7, and 21 after infection with 5 × 105 STm SL3261. Each point represents a single mouse (data are combined from 3 independent experiments); *P < .05 by 1-way ANOVA. (F) Line graph showing the level of thrombosis in the spleen (red line) and the liver (blue line) over the first 3 weeks of infection with 5 × 105 STm SL3261. The data are expressed as mean ± standard error of the mean from at least 4 mice per group combined from 3 independent experiments. (G) Quantification of thrombi in spleen sections from phosphate-buffered saline (PBS) liposomes or clodronate-liposome–pretreated mice infected for 24 hours with 5 × 105 STm SL3261. Combined data from 2 experiments with a total of 8 mice in each group are shown; *P < .05, ***P < .001 by 2-tailed nonparametric t test. N.D., not detected.

Thrombosis in the spleen and liver follows different kinetics after STm infection. (A) Frozen spleens from WT mice infected with 5 × 105 STm intraperitoneally (IP) were sectioned longitudinally to the hilum (≥1200 μm deep), and 5-μm sections were stained by IHC. Scans of stained spleen sections from mice infected for 0, 1, 7, and 21 days show blood vessels identified with anti-CD31 in blue and thrombi with antifibrin/fibrinogen in brown. Arrows identify individual thrombi. (B) Representative low power images of spleen and liver sections stained for CD41 (platelets; blue) and fibrin/fibrinogen (brown) from mice infected for 24 hours via the IV route with 5 × 105 STm SL3261. Arrows identify individual thrombi. The left-hand graph shows the number of thrombi per spleen section for mice infected IP or IV. The right-hand graph shows the number of thrombi per section in the spleen and liver from these IV-infected mice. (C) Representative low-power images of spleen and liver sections stained for CD41 (platelets; blue) and fibrin/fibrinogen (brown) from mice infected for 24 hours with the 105 virulent STm SL1344 strain. Arrows identify individual thrombi. The left-hand graph shows the number of thrombi per spleen section for mice infected with SL3261 (attenuated) or SL1344 (virulent). The right-hand graph shows the number of thrombi per section in the spleen and liver from mice infected with SL1344. (D) Representative scans at higher magnification of spleen sections (top) and liver sections (bottom) from WT mice at days 0, 1, 7, and 21 after infection with 5 × 105 STm SL3261. Sections are stained for fibrin/fibrinogen (brown) and CD41 (blue). RP, red pulp; T, thrombus; V, vein; WP, white pulp. (E) Quantification of numbers of thrombi per spleen section (left graph) and the proportion of section area covered by thrombi (right graph) at days 0, 1, 2, 7, and 21 after infection with 5 × 105 STm SL3261. Each point represents a single mouse (data are combined from 3 independent experiments); *P < .05 by 1-way ANOVA. (F) Line graph showing the level of thrombosis in the spleen (red line) and the liver (blue line) over the first 3 weeks of infection with 5 × 105 STm SL3261. The data are expressed as mean ± standard error of the mean from at least 4 mice per group combined from 3 independent experiments. (G) Quantification of thrombi in spleen sections from phosphate-buffered saline (PBS) liposomes or clodronate-liposome–pretreated mice infected for 24 hours with 5 × 105 STm SL3261. Combined data from 2 experiments with a total of 8 mice in each group are shown; *P < .05, ***P < .001 by 2-tailed nonparametric t test. N.D., not detected.

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