Journal: Scientific Reports
Article Title: Identification of Tp0751 (Pallilysin) as a Treponema pallidum Vascular Adhesin by Heterologous Expression in the Lyme disease Spirochete
doi: 10.1038/s41598-017-01589-4
Figure Lengend Snippet: Interaction of Tp0751-expressing strains with human endothelia under static and shear stress conditions. ( a ) Mean ± SEM numbers of parent and Tp0751-expressing B . burgdorferi ( Bb -Tp0751) adhered to primary human umbilical vein endothelial cell (HUVEC) monolayers after 12 h of co-incubation under static (0 dyn/cm 2 ) conditions (HUVECs + Bb , control). HUVECs+ ( Bb + pFn): binding of bacteria pre-incubated with plasma fibronectin (pFn) to HUVECs. (HUVECs + pFn) + Bb : binding of bacteria to HUVEC pre-incubated with pFn. N = 3 independent bacterial and endothelial cultures/strain analyzed in 2 experiments. ( b ) Mean ± SEM bacterial interactions per minute with HUVEC under typical postcapillary venule shear stress condition (1 dyn/cm 2 ). Videos of representative flow chamber experiments are presented in Supplementary Videos – . Numbers of bacteria that paused and moved more slowly over endothelia under flow were manually counted in time-lapses acquired at 15 frames per second (fps). Strains: Negative control strain: BBK32-deficient non-infectious adhesion-attenuated B31-A-derived strain (Parent; GCB706: Supplementary Table ). Positive control strains: Parent expressing B . burgdorferi vascular adhesin BBK32 ( Bb -BBK32); BBK32-expressing B31-derived infectious strain (Infectious; GCB726: Supplementary Table ). Bb -Tp0751: Parent expressing Tp0751. N = 8 independent cultures per strain analyzed in 3 experiments. Statistics: one-way Kruskal-Wallis ANOVA with Dunn’s post-test ( a ), one-way ANOVA with Holm-Sidak post-test ( b ). *Indicates p < 0.05 vs. parent within incubation condition ( a ) and vs. parent ( b ).
Article Snippet: For competitive inhibition experiments, bacteria or HUVEC were pre-incubated with 8 μg of plasma fibronectin (pFn) (Sigma, Oakville, Canada) for 1 hour.
Techniques: Expressing, Incubation, Binding Assay, Negative Control, Derivative Assay, Positive Control
