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Dengue virus NS1 cytokine-independent vascular leak is dependent on endothelial glycocalyx components

Fig 4

DENV2 NS1-induced vascular leak is independent of TLR4 and TNF-α signaling in vivo.

(A-B) Hair was removed from the dorsal dermis of (A-B) wild-type (n = 7), (A) Tlr4-/- (n = 3) and (B) TNF-αR-/- (n = 7) B6 mice, and mice were allowed to recover for 3 days. On the day of the assay, retro-orbital injections of Alexa Fluor 680-conjugated dextran were administered, followed by intradermal injections of PBS (black circles), 200 ng VEGF (purple squares), 15 μg DENV2 NS1 (blue triangles), and 7.5 μg DENV2 NS1 (green triangles). The dermis from each mouse was collected and processed two hours post-injection. Data represent quantification of mean fluorescent intensity from the dermis +/- SEM and were collected from 2–3 independent experiments. The same wild-type B6 mice are used for comparison in both (A) and (B). An ordinary one-way ANOVA with multiple comparisons to the PBS group using Dunnett’s multiple comparison test was used to determine significance of VEGF and DENV2 NS1. An unpaired, parametric, two-tailed t-test was used to determine significance between wild-type and knockout mice for each treatment group. ns = not significant, *P < 0.05, **P < 0.01, ****P < 0.0001.

Fig 4