“…An extended explanation of these mechanical variables can be found elsewhere. 1 Figure 6 HMEC-1 monolayer stresses pre- and post-exposure to Bb-GFP (A) Rows show representative: overlay of phase contrast image of HMEC-1 and Bb-GFP fluorescence (green); monolayer tensile stresses ( ); and compressive stresses ( ), for HMEC-1 in monolayer at different time points (columns) pre- and post-exposure to Bb-GFP. MOI is 200 bacteria per cell and cells reside on 3 kPa ECM.…”
Section: Step-by-step Methods Detailsmentioning
confidence: 99%
“…This is due in that in our original paper, we initially placed ECs on collagen I-coated glass coverslips and followed them through microscopy prior- and post-exposure to Bb to evidence that shortly after exposure to pathogens ECs lower their migration speed but EC speed is recovered at later times. 1 Triggered by the observation that EC kinematics displayed time-dependent alterations post-exposure to Bb , we sought to investigate whether those are due to changes in cellular force transduction and thus performed TFM by placing ECs on soft 3 kPa hydrogels. We found that they do.…”
“…An extended explanation of these mechanical variables can be found elsewhere. 1 Figure 6 HMEC-1 monolayer stresses pre- and post-exposure to Bb-GFP (A) Rows show representative: overlay of phase contrast image of HMEC-1 and Bb-GFP fluorescence (green); monolayer tensile stresses ( ); and compressive stresses ( ), for HMEC-1 in monolayer at different time points (columns) pre- and post-exposure to Bb-GFP. MOI is 200 bacteria per cell and cells reside on 3 kPa ECM.…”
Section: Step-by-step Methods Detailsmentioning
confidence: 99%
“…This is due in that in our original paper, we initially placed ECs on collagen I-coated glass coverslips and followed them through microscopy prior- and post-exposure to Bb to evidence that shortly after exposure to pathogens ECs lower their migration speed but EC speed is recovered at later times. 1 Triggered by the observation that EC kinematics displayed time-dependent alterations post-exposure to Bb , we sought to investigate whether those are due to changes in cellular force transduction and thus performed TFM by placing ECs on soft 3 kPa hydrogels. We found that they do.…”
“…In the literature, there are numerous studies showing that cells affected by a disease exhibit different nanomechanical properties [ 288 ] and that cell elasticity is a critical determinant of proper cell function [ 289 ]. Specifically in LD Borrelia , exposure of endothelial cells to spirochaetes led to decreased motility and physical forces in host cell monolayers and affected the mechanotransduction of the endothelial cells [ 25 ]. An understanding of the structural rearrangements and mechanical responses of human cells to pathogen infection may shed light on the origin of structure-related clinical manifestations such as skin fibrosis, morphea, or hyperkeratosis.…”
Section: Alternative Pathogenic Mechanismsmentioning
Infection with
Borrelia burgdorferi
often triggers pathophysiologic perturbations that are further augmented by the inflammatory responses of the host, resulting in the severe clinical conditions of Lyme disease. While our apprehension of the spatial and temporal integration of the virulence determinants during the enzootic cycle of
B. burgdorferi
is constantly being improved, there is still much to be discovered. Many of the novel virulence strategies discussed in this review are undetermined. Lyme disease spirochaetes must surmount numerous molecular and mechanical obstacles in order to establish a disseminated infection in a vertebrate host. These barriers include borrelial relocation from the midgut of the feeding tick to its body cavity and further to the salivary glands, deposition to the skin, haematogenous dissemination, extravasation from blood circulation system, evasion of the host immune responses, localization to protective niches, and establishment of local as well as distal infection in multiple tissues and organs. Here, the various well-defined but also possible novel strategies and virulence mechanisms used by
B. burgdorferi
to evade obstacles laid out by the tick vector and usually the mammalian host during colonization and infection are reviewed.
“…Incubation with rabbit polyclonal B. burgdorferi antibodies specific to a pool of B. burgdorferi s.l. complex proteins (cat.# PA1-73004, Invitrogen, Carlsbad, CA, USA) diluted at 1:200 in the blocking solution was carried out in a humidified chamber overnight at 4 • C, followed by washing the samples by thorough rinsing with PBS-Tw (three times for 10 min at RT) [118,119]. For enzymatic staining, the sections were further incubated in the cross-adsorbed horseradish peroxidase-labeled goat anti-rabbit secondary antibodies (Invitrogen, Carlsbad, USA) diluted at 1:500 in the blocking solution for 90 min in RT, washed in PBS-Tw (three times for 10 min at RT), in 0.05 M Tris-HCl pH 7.5 (for 10 min at RT), and stained in 10% 3,3' diaminobenzidine in 0.05 M Tris-HCl pH 7.5 with 0.005% H 2 O 2 for 10 min at RT.…”
Section: Immunohistochemical Detection Of Borrelia In the Paraplast S...mentioning
Lyme disease (LD) spirochetes are well known to be able to disseminate into the tissues of infected hosts, including humans. The diverse strategies used by spirochetes to avoid the host immune system and persist in the host include active immune suppression, induction of immune tolerance, phase and antigenic variation, intracellular seclusion, changing of morphological and physiological state in varying environments, formation of biofilms and persistent forms, and, importantly, incursion into immune-privileged sites such as the brain. Invasion of immune-privileged sites allows the spirochetes to not only escape from the host immune system but can also reduce the efficacy of antibiotic therapy. Here we present a case of the detection of spirochetal DNA in multiple loci in a LD patient’s post-mortem brain. The presence of co-infection with Borrelia burgdorferi sensu stricto and Borrelia garinii in this LD patient’s brain was confirmed by PCR. Even though both spirochete species were simultaneously present in human brain tissue, the brain regions where the two species were detected were different and non-overlapping. The presence of atypical spirochete morphology was noted by immunohistochemistry of the brain samples. Atypical morphology was also found in the tissues of experimentally infected mice, which were used as a control.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
