scholarly journals Listeriolysin O Suppresses Phospholipase C-Mediated Activation of the Microbicidal NADPH Oxidase to Promote Listeria monocytogenes Infection

2011 ◽  
Vol 10 (6) ◽  
pp. 627-634 ◽  
Author(s):  
Grace Y. Lam ◽  
Ramzi Fattouh ◽  
Aleixo M. Muise ◽  
Sergio Grinstein ◽  
Darren E. Higgins ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Nadph Oxidase ◽  
Phospholipase C ◽  
Listeriolysin O ◽  
Listeria Monocytogenes Infection

scholarly journals Listeria monocytogenes Phospholipase C-Dependent Calcium Signaling Modulates Bacterial Entry into J774 Macrophage-Like Cells

1999 ◽  
Vol 67 (4) ◽  
pp. 1770-1778 ◽  
Author(s):  
Sandra J. Wadsworth ◽  
Howard Goldfine
Keyword(s):  
Listeria Monocytogenes ◽  
Calcium Signaling ◽  
Phospholipase C ◽  
Virulence Factors ◽  
Intracellular Signaling ◽  
Lethal Dose ◽  
Listeriolysin O ◽  
Wild Type ◽  
J774 Macrophage ◽  
Kinetics Of

ABSTRACT Listeria monocytogenes secretes several proteins that have been shown to contribute to virulence. Among these is listeriolysin O (LLO), a pore-forming hemolysin that is absolutely required for virulence. Two other virulence factors are phospholipases: a phosphatidylinositol-specific phospholipase C (PI-PLC [plcA]) and a broad-range PLC (plcB). Although mutations in plcA or plcB resulted in small increases in mouse 50% lethal dose (LD50), deletions in both genes resulted in a 500-fold increase in LD50. We have examined the role of these secreted proteins in host intracellular signaling in the J774 macrophage-like cell line. Measurements of cytosolic free calcium ([Ca2+]i) have revealed a rapid spike upon exposure of these cells to wild-typeL. monocytogenes. This is followed by a second peak at 5 min and a third prolonged peak with a maximal [Ca2+]i of 800 to 1,000 nM. The pattern of calcium changes was greatly altered by deletion of any of the three virulence factors. An LLO mutant produced none of these elevations in [Ca2+]i; however, a transient elevation was observed whenever these bacteria entered the cell. A PI-PLC mutant produced a diminished single elevation in [Ca2+]i at 15 to 30 min. A broad-range PLC mutant produced only the first calcium spike. Studies with inhibitors suggested that the first elevation arises from influx of calcium from the extracellular medium through plasma membrane channels and that the second and third elevations come from release of Ca2+ from intracellular stores. We observed that internalization of wild-type bacteria and the broad-range PLC mutant was delayed for 5 to 10 min, but the LLO and PI-PLC mutants were internalized rapidly upon infection. Inhibitors that affected calcium signaling changed the kinetics of association of wild-type bacteria with J774 cells, the kinetics of entry, and the efficiency of escape from the primary phagosome.

scholarly journals Listeriolysin O Is Necessary and Sufficient to Induce Autophagy during Listeria monocytogenes Infection

PLoS ONE ◽  
2010 ◽  
Vol 5 (1) ◽  
pp. e8610 ◽  
Author(s):  
Nicole Meyer-Morse ◽  
Jennifer R. Robbins ◽  
Chris S. Rae ◽  
Sofia N. Mochegova ◽  
Michele S. Swanson ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Listeriolysin O ◽  
Listeria Monocytogenes Infection ◽  
Necessary And Sufficient

scholarly journals Activation of Host Phospholipases C and D in Macrophages after Infection with Listeria monocytogenes

2000 ◽  
Vol 68 (10) ◽  
pp. 5735-5741 ◽  
Author(s):  
Howard Goldfine ◽  
Sandra J. Wadsworth ◽  
Norah C. Johnston
Keyword(s):  
Listeria Monocytogenes ◽  
Intracellular Calcium ◽  
Phospholipase C ◽  
Term Treatment ◽  
Listeriolysin O ◽  
Wild Type ◽  
Murine Bone Marrow ◽  
Long Term Treatment ◽  
J774 Cells ◽  
Hydrolysis Of

ABSTRACT Infection of the J774 murine macrophage-derived cell line withListeria monocytogenes results in several elevations of intracellular calcium during the first 15 min of infection. These appear to result from the actions of secreted bacterial proteins, including phosphatidylinositol-specific phospholipase C (PI-PLC), a broad-range phospholipase C, and listeriolysin O (LLO) (S. J. Wadsworth and H. Goldfine, Infect. Immun. 67:1770–1778, 1999). We have measured hydrolysis of host PI and the activation of host polyphosphoinositide-specific PLC and host phospholipase D (PLD) during infection with wild-type and mutant L. monocytogenes. Elevated hydrolysis of host PI occurred within the first 10 min of infection and was dependent on both bacterial PI-PLC and LLO, both of which were required for the earliest elevations of intracellular calcium in the host cell. A more rapid hydrolysis of host PI was observed at 30 min after infection, at the time when wild-type bacteria have been internalized. Activation of host PLC, also occurred in the first 10 min of infection but was not dependent on the presence of bacterial PI-PLC. Similar observations were made in murine bone marrow-derived macrophages. In J774 cells, activation of host PLD was observed after 20 min of infection and was dependent on bacterial LLO. Mutants in the bacterial phospholipases produced levels of PLD activation similar to those produced by the wild type. Phorbol myristate acetate (PMA) also activated host PLD, while long-term treatment with PMA resulted in loss of the ability of L. monocytogenes to activate host PLD, suggesting an involvement of protein kinase C (PKC) in the activation of PLD. Rottlerin, an inhibitor of PKC δ in J774 cells, also inhibited the activation of PLD, but hispidin, an inhibitor of PKC βI and βII, did not. Pretreatment of J774 cells with the PLD inhibitor, 2,3-diphosphoglycerate partially inhibited escape of the bacteria from the primary phagocytic vacuole.

scholarly journals Passive immunization with anti-ActA and anti-listeriolysin O antibodies protects against Listeria monocytogenes infection in mice

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Krisana Asano ◽  
Hiroshi Sashinami ◽  
Arihiro Osanai ◽  
Shouhei Hirose ◽  
Hisaya K. Ono ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Passive Immunization ◽  
Listeriolysin O ◽  
Listeria Monocytogenes Infection

scholarly journals Requirement of the Listeria monocytogenes Broad-Range Phospholipase PC-PLC during Infection of Human Epithelial Cells

2003 ◽  
Vol 185 (21) ◽  
pp. 6295-6307 ◽  
Author(s):  
Angelika Gründling ◽  
Mark D. Gonzalez ◽  
Darren E. Higgins
Keyword(s):  
Listeria Monocytogenes ◽  
Epithelial Cells ◽  
Host Cell ◽  
Phospholipase C ◽  
Expression System ◽  
Broth Culture ◽  
Listeriolysin O ◽  
Proteolytic Activation ◽  
Human Epithelial Cells ◽  
Cell Spread

ABSTRACT In this study, we investigated the requirement of the Listeria monocytogenes broad-range phospholipase C (PC-PLC) during infection of human epithelial cells. L. monocytogenes is a facultative intracellular bacterial pathogen of humans and a variety of animal species. After entering a host cell, L. monocytogenes is initially surrounded by a membrane-bound vacuole. Bacteria promote their escape from this vacuole, grow within the host cell cytosol, and spread from cell to cell via actin-based motility. Most infection studies with L. monocytogenes have been performed with mouse cells or an in vivo mouse model of infection. In all mouse-derived cells tested, the pore-forming cytolysin listeriolysin O (LLO) is absolutely required for lysis of primary vacuoles formed during host cell entry. However, L. monocytogenes can escape from primary vacuoles in the absence of LLO during infection of human epithelial cell lines Henle 407, HEp-2, and HeLa. Previous studies have shown that the broad-range phospholipase C, PC-PLC, promotes lysis of Henle 407 cell primary vacuoles in the absence of LLO. Here, we have shown that PC-PLC is also required for lysis of HEp-2 and HeLa cell primary vacuoles in the absence of LLO expression. Furthermore, our results indicated that the amount of PC-PLC activity is critical for the efficiency of vacuolar lysis. In an LLO-negative derivative of L. monocytogenes strain 10403S, expression of PC-PLC has to increase before or upon entry into human epithelial cells, compared to expression in broth culture, to allow bacterial escape from primary vacuoles. Using a system for inducible PC-PLC expression in L. monocytogenes, we provide evidence that phospholipase activity can be increased by elevated expression of PC-PLC or Mpl, the enzyme required for proteolytic activation of PC-PLC. Lastly, by using the inducible PC-PLC expression system, we demonstrate that, in the absence of LLO, PC-PLC activity is not only required for lysis of primary vacuoles in human epithelial cells but is also necessary for efficient cell-to-cell spread. We speculate that the additional requirement for PC-PLC activity is for lysis of secondary double-membrane vacuoles formed during cell-to-cell spread.

Listeria monocytogeneslisteriolysin O and phosphatidylinositol-specific phospholipase C affect adherence to epithelial cells

2005 ◽  
Vol 51 (9) ◽  
pp. 745-751 ◽  
Author(s):  
Agata Krawczyk-Balska ◽  
Jacek Bielecki
Keyword(s):  
Listeria Monocytogenes ◽  
Epithelial Cells ◽  
Phospholipase C ◽  
Listeriolysin O ◽  
Cell Responses ◽  
Bacterial Binding ◽  
Intracellular Ca ◽  
Infected Cells ◽  
Host Parasite ◽  
Phagosomal Membrane

Listeria monocytogenes, a foodborn intracellular animal and human pathogen, produces several exotoxins contributing to virulence. Among these are listeriolysin O (LLO), a pore-forming cholesterol-dependent hemolysin, and a phosphatidylinositol-specific phospholipase C (PI-PLC). LLO is known to play an important role in the escape of bacteria from the primary phagocytic vacuole of macrophages, and PI-PLC supports this process. Evidence is accumulating that LLO and PI-PLC are multifunctional virulence factors with many important roles in the host-parasite interaction other than phagosomal membrane disruption. LLO and PI-PLC may induce a number of host cell responses by modulating signal transduction of infected cells via intracellular Ca2+levels and the metabolism of phospholipids. This would result in the activation of host phospholipase C and protein kinase C. In the present study, using Bacillus sub tilis strains expressing LLO, PI-PLC, and simultaneously LLO and PI-PLC, we show that LLO and PI-PLC enhance bacterial binding to epithelial cells Int407, with LLO being necessary and PI-PLC playing an accessory role. The results of this work suggest that these two listerial proteins act on epithelial cells prior to internalization.Key words: Listeria monocytogenes, listeriolysin O, phosphatidylinositol-specific phospholipase C, Bacillus subtilis, adherence.

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