Neisseria meningitidis Induces Brain Microvascular Endothelial Cell Detachment from the Matrix and Cleavage of Occludin: A Role for MMP-8
Figure 7
N. meningitidis infection triggers caspase-independent detachment of HBMEC.
(A) Confluent monolayers of HBMEC were infected with N. meningitidis MC58 and the unencapsulated isogenic mutant strain MC58 siaD for a 24-hour period in the presence of the pan-caspase inhibitor Z-VAD-fmk. Cells were used in detachment assays and the percentage of detached cells was determined and is expressed as a percentage (mean ± SD of two wells from five independent assays) relative to detachment observed in uninfected control cells. Error bars represent means ± SD of three independent experiments. (B) HBMEC were incubated with MC58 and MC58 siaD (MOI 30) and MMP-8 activity was measured in supernatants collected from infected HBMEC at 2, 4, 8, and 24 h p.i. using the Sensolyte 520 MMP-8 assay kit. (C) HBMEC were treated with 1 µM and 5 µM Staurosporin (STS) for 2, 4 and 8 h and MMP-8 activity was measured in the supernatant of STS-treated cells. * P<0.05.