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. 2017 May 8;7:1538. doi: 10.1038/s41598-017-01589-4

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Effect of Tp0751 on tethered and untethered interaction stability and velocity under shear stress. B. burgdorferi interacts with endothelia under PCV shear stress conditions by two mechanisms: untethered (dragging) interactions, and tethered interactions. Tethered interactions are stabilized by bungee cord-like tethers which anchor bacteria to endothelia, reduce force imposed on load-bearing adhesion complexes and increase adhesion bond stability (lifetime)²⁹. (a,b) Proportion of total bacterial-endothelial interaction times during which bacterial length extended beyond the normal length, indicative of tether formation stabilizing the interaction ((a) % duration), and the average tether length (b) for all interactions at 0.5–1 dyn/cm². (c,d) Interaction complex dissociation rates for untethered (c) and tethered (d) interactions. Dotted lines indicate dissociation rates for non-specific interactions (non-adherent beads). (e,f) Velocities of untethered (e) and tethered (f) interactions. Summary values: mean ± 95% CI (a,b,e,f), mean ± SEM (c,d). Interaction properties were calculated from interactions obtained at both 0.5 and 1 dyn/cm². N = 4 and N = 8 independent flow chamber experiments for each strain at 0.5 and 1 dyn/cm², respectively. Strains: Negative control strain: BBK32-deficient non-infectious adhesion-attenuated B31-A-derived strain (Parent; GCB706: Supplementary Table S1). Positive control strains: Parent expressing B. burgdorferi vascular adhesin BBK32 (Bb-BBK32); BBK32-expressing B31-derived infectious strain (Infectious; GCB726: Supplementary Table S1). Bb-Tp0751: Parent expressing Tp0751. Numbers of interactions analyzed/strain (a,b): ≥288; (c,e): parent (42), infectious (127), Bb-BBK32 (130) and Bb-Tp0751 (39); (d,f): parent (366), infectious (520), Bb-BBK32 (448), and Bb-Tp0751 (249). Statistics: one-way Kruskal-Wallis ANOVA with Dunn’s post-tests (a,b,e,f); one-way ANOVA with Holm-Sidak post-tests (c,d). *Indicates p < 0.05 vs parent. ⁺Indicates p < 0.05 vs Bb-BBK32.