Use of CFSE staining of borreliae in studies on the interaction between borreliae and human neutrophils

Tuominen-Gustafsson, Helena; Penttinen, Markus A.; Hytönen, Jukka; Viljanen, Matti K.

doi:10.1186/1471-2180-6-92

Cited by 39 publications

(26 citation statements)

References 36 publications

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“…Previous work showed that macrophages, dendritic cells, and neutrophils can phagocytose and kill B. burgdorferi in vitro using conventional, coiling, or tube phagocytosis (9,(11)(12)(13)(14)(15)(16)(17)(18). Additional studies examined the in vivo role of neutrophils in controlling B. burgdorferi infection at the inoculation site and showed that increased neutrophil recruitment to the site of inoculation of engineered B. burgdorferi severely attenuates B. burgdorferi infectivity (19).…”

Section: Discussionmentioning

confidence: 99%

“…These results suggest that neutrophils have a limited role in controlling B. burgdorferi infection in mice in the conventional laboratory model system tested. Therefore, while macrophages, dendritic cells, and neutrophils have been shown to phagocytose B. burgdorferi in vitro using conventional, coiling, or tube phagocytosis (9,(11)(12)(13)(14)(15)(16)(17)(18), none of these innate immune effector cell types individually appears to have a major role in controlling B. burgdorferi infection in mice under conditions tested here or in our previous work (43). One caveat is that these cells types may have redundant phagocytic functions, so it is possible that one cell type can compensate for depletion of another in these experiments.…”

Section: Discussionmentioning

confidence: 99%

“…interactions between B. burgdorferi and the cellular infiltrates has been the subject of several previous studies. Macrophages, dendritic cells, and neutrophils have been shown to phagocytose and kill B. burgdorferi in vitro through conventional, coiling, or tube phagocytosis (9,(11)(12)(13)(14)(15)(16)(17)(18), leading to the hypothesis that each of these innate immune cells participates in control of early B. burgdorferi infections in vivo. Previous studies have tested this hypothesis by examining the in vivo role of neutrophils in controlling B. burgdorferi infection in mice.…”

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confidence: 99%

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Characterization of Stress and Innate Immunity Resistance of Wild-Type and Δp66 Borrelia burgdorferi

et al. 2018

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is a causative agent of Lyme disease, the most common arthropod-borne disease in the United States. evades host immune defenses to establish a persistent, disseminated infection. Previous work showed that P66-deficient (Δ) is cleared quickly after inoculation in mice. We demonstrate that the Δ strain is rapidly cleared from the skin inoculation site prior to dissemination. The rapid clearance of Δ bacteria is not due to inherent defects in multiple properties that might affect infectivity: bacterial outer membrane integrity, motility, chemotactic response, or nutrient acquisition. This led us to the hypothesis that P66 has a role in mouse cathelicidin-related antimicrobial peptide (mCRAMP; a major skin antimicrobial peptide) and/or neutrophil evasion. Neither wild-type (WT) nor Δ was susceptible to mCRAMP. To examine the role of neutrophil evasion, we administered neutrophil-depleting antibody anti-Ly6G (1A8) to C3H/HeN mice and subsequently monitored the course of infection. Δ mutants were unable to establish infection in neutrophil-depleted mice, suggesting that the important role of P66 during early infection is through another mechanism. Neutrophil depletion did not affect WT bacterial burdens in the skin (inoculation site), ear, heart, or tibiotarsal joint at early time points postinoculation. This was unexpected given that prior studies demonstrated neutrophils phagocytose and kill These data, together with our previous work, suggest that despite the ability of host innate defenses to kill , individual innate immune mechanisms have limited contributions to controlling early infection in the laboratory model used.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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Characterization of Stress and Innate Immunity Resistance of Wild-Type and Δp66 Borrelia burgdorferi

et al. 2018

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“…To investigate whether S. rugosus and S. rotundus are intracellular bacteria, we used carboxyfluorescein diacetate succinimidyl ester (CFSE; Invitrogen, Carlsbad, CA, USA) for staining of bacteria as previously described [14]. Briefly, 1×10 8 cells/ml of bacteria were washed once in phosphate buffered saline (PBS) and diluted in 500 µl of PBS.…”

Section: Methodsmentioning

confidence: 99%

Differential Immune Responses to Segniliparus rotundus and Segniliparus rugosus Infection and Analysis of Their Comparative Virulence Profiles

Kim

Lee

et al. 2013

PLoS ONE

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Two closely related bacterial species, Segniliparus rotundus and Segniliparus rugosus, have emerged as important human pathogens, but little is known about the immune responses they elicit or their comparative pathophysiologies. To determine the virulence and immune responses of the two species, we compared their abilities to grow in phagocytic and non-phagocytic cells. Both species maintained non-replicating states within A549 epithelial cells. S. rugosus persisted longer and multiplied more rapidly inside murine bone marrow-derived macrophages (BMDMs), induced more pro-inflammatory cytokines, and induced higher levels of macrophage necrosis. Activation of BMDMs by both species was mediated by toll-like receptor 2 (TLR2), followed by mitogen-activated protein kinases (MAPK) and nuclear factor κB (NF-κB) signaling pathways, indicating a critical role for TLR2 in Segniliparus-induced macrophage activation. S. rugosus triggered faster and stronger activation of MAPK signaling and IκB degradation, indicating that S. rugosus induces more pro-inflammatory cytokines than S. rotundus. Multifocal granulomatous inflammations in the liver and lung were observed in mice infected with S. rugosus, but S. rotundus was rapidly cleared from all organs tested within 15 days post-infection. Furthermore, S. rugosus induced faster infiltration of innate immune cells such as neutrophils and macrophages to the lung than S. rotundus. Our results suggest that S. rugosus is more virulent and induces a stronger immune response than S. rotundus.

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“…Erythromycin (5 g/ml) and tetracycline (2 g/ml) were added according to the selection requirements of the strains (47). Bacteria were washed once in phosphate-buffered saline (PBS) and stained with carboxyfluorescein succinimidyl ester (CFSE) (eBioscience) as described for flow cytometry (48). Briefly, bacteria in PBS were stained with CFSE at a final concentration of 5 M for 30 min at 37°C and protected from light.…”

Section: Methodsmentioning

confidence: 99%

Secondary Lymphoid Organ Homing Phenotype of Human Myeloid Dendritic Cells Disrupted by an Intracellular Oral Pathogen

et al. 2014

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c Several intracellular pathogens, including a key etiological agent of chronic periodontitis, Porphyromonas gingivalis, infect blood myeloid dendritic cells (mDCs). This infection results in pathogen dissemination to distant inflammatory sites (i.e., pathogen trafficking). The alteration in chemokine-chemokine receptor expression that contributes to this pathogen trafficking function, particularly toward sites of neovascularization in humans, is unclear. To investigate this, we utilized human monocytederived DCs (MoDCs) and primary endothelial cells in vitro, combined with ex vivo-isolated blood mDCs and serum from chronic periodontitis subjects and healthy controls. Our results, using conditional fimbria mutants of P. gingivalis, show that P. gingivalis infection of MoDCs induces an angiogenic migratory profile. This profile is enhanced by expression of DC-SIGN on MoDCs and minor mfa-1 fimbriae on P. gingivalis and is evidenced by robust upregulation of CXCR4, but not secondary lymphoid organ (SLO)-homing CCR7. This disruption of SLO-homing capacity in response to respective chemokines closely matches surface expression of CXCR4 and CCR7 and is consistent with directed MoDC migration through an endothelial monolayer. Ex vivo-isolated mDCs from the blood of chronic periodontitis subjects, but not healthy controls, expressed a similar migratory profile; moreover, sera from chronic periodontitis subjects expressed elevated levels of CXCL12. Overall, we conclude that P. gingivalis actively "commandeers" DCs by reprogramming the chemokine receptor profile, thus disrupting SLO homing, while driving migration toward inflammatory vascular sites.

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Use of CFSE staining of borreliae in studies on the interaction between borreliae and human neutrophils

Cited by 39 publications

References 36 publications

Characterization of Stress and Innate Immunity Resistance of Wild-Type and Δp66 Borrelia burgdorferi

Characterization of Stress and Innate Immunity Resistance of Wild-Type and Δp66 Borrelia burgdorferi

Differential Immune Responses to Segniliparus rotundus and Segniliparus rugosus Infection and Analysis of Their Comparative Virulence Profiles

Secondary Lymphoid Organ Homing Phenotype of Human Myeloid Dendritic Cells Disrupted by an Intracellular Oral Pathogen

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