Capacity of listeriolysin O, streptolysin O, and perfringolysin O to mediate growth of Bacillus subtilis within mammalian cells

Portnoy, Daniel A.; Tweten, Rodney K.; Kehoe, Michael; Bielecki, Jacek

doi:10.1128/iai.60.7.2710-2717.1992

Cited by 114 publications

(58 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…To determine if LLO was the only L. monocytogenes determinant of pathogenesis necessary to induce autophagy, we examined the interaction of LC3-GFP in BMDMs with the non-pathogenic Gram-positive bacterium Bacillus subtilis engineered to express and secrete LLO or a related cholesterol dependent cytolysin, perfringolysin O (PFO). These modified bacteria have previously been shown to escape into the host cell cytosol, although the efficiency of escape is less than that observed for wild-type L. monocytogenes [29,30].…”

Section: Llo and Pfo Expression By Bacillus Subtilis Induced Bacteriamentioning

confidence: 88%

See 1 more Smart Citation

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

et al. 2010

View full text Add to dashboard Cite

BackgroundRecent studies have suggested that autophagy is utilized by cells as a protective mechanism against Listeria monocytogenes infection.Methodology/Principal FindingsHowever we find autophagy has no measurable role in vacuolar escape and intracellular growth in primary cultured bone marrow derived macrophages (BMDMs) deficient for autophagy (atg5−/−). Nevertheless, we provide evidence that the pore forming activity of the cholesterol-dependent cytolysin listeriolysin O (LLO) can induce autophagy subsequent to infection by L. monocytogenes. Infection of BMDMs with L. monocytogenes induced microtubule-associated protein light chain 3 (LC3) lipidation, consistent with autophagy activation, whereas a mutant lacking LLO did not. Infection of BMDMs that express LC3-GFP demonstrated that wild-type L. monocytogenes was encapsulated by LC3-GFP, consistent with autophagy activation, whereas a mutant lacking LLO was not. Bacillus subtilis expressing either LLO or a related cytolysin, perfringolysin O (PFO), induced LC3 colocalization and LC3 lipidation. Further, LLO-containing liposomes also recruited LC3-GFP, indicating that LLO was sufficient to induce targeted autophagy in the absence of infection. The role of autophagy had variable effects depending on the cell type assayed. In atg5−/− mouse embryonic fibroblasts, L. monocytogenes had a primary vacuole escape defect. However, the bacteria escaped and grew normally in atg5−/− BMDMs.Conclusions/SignificanceWe propose that membrane damage, such as that caused by LLO, triggers bacterial-targeted autophagy, although autophagy does not affect the fate of wild-type intracellular L. monocytogenes in primary BMDMs.

show abstract

Section: Llo and Pfo Expression By Bacillus Subtilis Induced Bacteriamentioning

confidence: 88%

“…The wild-type Bacillus subtilis strain used was DP-B1066 [30]. It contains a vector-only insertion with no hemolysin.…”

Section: Bacterial Strainsmentioning

confidence: 99%

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

et al. 2010

View full text Add to dashboard Cite

show abstract

“…Listeriolysin O is essential for the escape of L. monocytogenes from the macrophage phagosome to the cytoplasmic compartment [3]. It has been shown previously that an engineered LLO-secreting Bacillus subtilis strain can escape from the phagosomal compartment and grow in the J774 cell cytosol [49]. In contrast, Goetz et al utilized microinjection to introduce a variety of bacteria directly into the cytoplasm of Caco-2 cells and found that bacteria that are not adapted to the cytoplasmic compartment are unable to multiply and grow [50].…”

Section: Discussionmentioning

confidence: 99%

Lactococcus lactis-expressing listeriolysin O (LLO) provides protection and specific CD8+ T cells against Listeria monocytogenes in the murine infection model

Bahey-El-Din

Casey

Griffin

et al. 2008

Vaccine

View full text Add to dashboard Cite

Lactococcus lactis has previously been proposed as a vaccine platform for the safe delivery of heterologous antigens. Here we utilized L. lactis as a live vector for expression of listeriolysin O (LLO), a major Listeria monocytogenes antigen and virulence factor. A variety of plasmid constructs were designed to permit either constitutive or nisin-inducible expression of secreted or non-secreted LLO in L. lactis. Recombinant strains were subsequently tested in a murine model for vaccination efficacy against L. monocytogenes infection. CD8(+) T lymphocytes specific for the LLO(91-99) epitope were detected when strains were administered via the intraperitoneal (IP) but not the oral route. Challenge with live L. monocytogenes revealed different levels of protection among the three vaccine strains tested with the nisin-inducible LLO-secreting L. lactis strain providing the greatest protection against secondary infection. This work highlights the usefulness of the GRAS (Generally Regarded As Safe) organism L. lactis as the basis of a live vaccine vector against L. monocytogenes. The work suggests that LLO-expressing L. lactis strains may also have the potential to act as a platform for directing other co-expressed antigens towards the cytosolic MHC class I pathway for enhanced stimulation of the CD8(+) T-cell response.

show abstract

“…Lm escape can be inhibited by alkalinization of the phagosome using bafilomycin A1, an inhibitor of vacuolar ATPases (Beauregard et al ., 1997). This may be due to the acidic pH optimum for cytolytic activity by LLO (Portnoy et al ., 1992), or to other pH-dependent processes that affect LLO function indirectly.…”

Section: Introductionmentioning

confidence: 99%

Membrane perforations inhibit lysosome fusion by altering pH and calcium in Listeria monocytogenes vacuoles

Shaughnessy¹,

Hoppe²,

Christensen³

et al. 2006

Cell Microbiol

147

140

View full text Add to dashboard Cite

SummaryListeria monocytogenes (Lm) evade microbicidal defences inside macrophages by secreting a poreforming cytolysin listeriolysin O (LLO), which allows Lm to escape vacuoles. LLO also inhibits Lm vacuole fusion with lysosomes, which indicates LLO alters vacuole chemistry prior to release of Lm into cytoplasm. Using fluorescent probes to measure membrane permeability, calcium and pH, we identified small membrane perforations in vacuoles containing wild-type but not LLO-deficient (hly-) Lm. The small membrane perforations released small fluorescent molecules and persisted for several minutes before expanding to allow exchange of larger fluorescent molecules. Macropinosomes and hly-Lm vacuoles acidified and increased their calcium content ([Ca 2+ ] vac ) within minutes of formation; however, the small perforations made by LLO-expressing bacteria increased vacuolar pH and decreased [Ca 2+ ] vac shortly after infection. Experimental increases in vacuolar pH inhibited Lm vacuole fusion with lysosomes. The timing of perforation indicated that LLO-dependent delays of Lm vacuole maturation result from disruption of ion gradients across vacuolar membranes.

show abstract

Capacity of listeriolysin O, streptolysin O, and perfringolysin O to mediate growth of Bacillus subtilis within mammalian cells

Cited by 114 publications

References 44 publications

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

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

Lactococcus lactis-expressing listeriolysin O (LLO) provides protection and specific CD8+ T cells against Listeria monocytogenes in the murine infection model

Membrane perforations inhibit lysosome fusion by altering pH and calcium in Listeria monocytogenes vacuoles

Contact Info

Product

Resources

About