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Liu, G. Y. et al. Sword and shield: Linked group B streptococcal -hemolysin/cytolysin and carotenoid pigment function to subvert host phagocyte defense. Proc. Natl Acad. Sci. USA 101, 14491-14496

Department of Pediatrics, The Scripps Institution of Oceanography, University of California at San Diego, La Jolla, CA 92093, USA.
Proceedings of the National Academy of Sciences (Impact Factor: 9.67). 11/2004; 101(40):14491-6. DOI: 10.1073/pnas.0406143101
Source: PubMed

ABSTRACT

Group B Streptococcus (GBS) is a major cause of pneumonia, bacteremia, and meningitis in neonates and has been found to persist inside host phagocytic cells. The pore-forming GBS beta-hemolysin/cytolysin (betaH/C) encoded by cylE is an important virulence factor as demonstrated in several in vivo models. Interestingly, cylE deletion results not only in the loss of betaH/C activity, but also in the loss of a carotenoid pigment of unknown function. In this study, we sought to define the mechanism(s) by which cylE may contribute to GBS phagocyte resistance and increased virulence potential. We found that cylE-deficient GBS was more readily cleared from a mouse's bloodstream, human whole blood, and isolated macrophage and neutrophil cultures. Survival was linked to the ability of betaH/C to induce cytolysis and apoptosis of the phagocytes. At a lower bacterial inoculum, cylE also contributed to enhanced survival within phagocytes that was attributed to the ability of carotenoid to shield GBS from oxidative damage. In oxidant killing assays, cylE mutants were shown to be more susceptible to hydrogen peroxide, hypochlorite, superoxide, and singlet oxygen. Together, these data suggest a mechanism by which the linked cylE-encoded phenotypes, betaH/C (sword) and carotenoid (shield), act in partnership to thwart the immune phagocytic defenses.

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Available from: Kelly Doran, Sep 24, 2015
    • "The remaining 20 strains have independent mutations in at least 1 of the 12 cyl operon genes. While cylE inactivation has been shown to systematically yield NH/NP mutants in several unrelated isolates, the phenotype resulting from the inactivation of the other cyl genes had not been systematically studied (11, 18, 19, 21, 36). Seven of the 12 genes of the cyl operon are thought to be involved in the synthesis of the lipid backbone of the carotenoid-like molecule granadaene (cylX, cylD, cylG, acpC, cylZ, cylI, and cylK) (11, 18, 19, 21, 36). "
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    ABSTRACT: Group B Streptococcus (GBS) is a common commensal bacterium in adults, but is also the leading cause of invasive bacterial infections in neonates in developed countries. The ß-haemolysin/cytolysin (ß-h/c), which is always associated with the production of an orange-to-red pigment, is a major virulence factor that is also used for GBS diagnosis. A collection of 1,776 independent clinical GBS strains isolated in France between 2006 and 2013 was evaluated on specific medium for ß-h/c activity and pigment production. Genomic sequences of non-haemolytic and non-pigmented (NH/NP) strains were analyzed to identify the molecular basis of this phenotype. Gene deletions or complementations were carried out to confirm the genotype-phenotype association. Sixty-three GBS strains (3.5%) were NH/NP and 47 of these (74.6%) originate from invasive infections, including bacteremia and meningitis, in neonates or adults. Mutations are localized predominantly in the cyl operon, encoding the ß-h/c-pigment biosynthetic pathway, and in the abx1 gene, encoding a CovSR regulator partner. In conclusion, although usually associated with GBS virulence, ß-h/c-pigment production is not absolutely required to cause human invasive infections. Caution should therefore be taken when using hemolysis and pigmentation as criteria for GBS diagnosis in routine clinical laboratory settings.
    No preview · Article · Oct 2015 · Journal of clinical microbiology
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    • "This toxin is responsible for the characteristic zone of b-hemolysis exhibited by GBS, and hemolytic strains are associated with virulence (Liu et al, 2004; Fettucciari et al, 2006; Costa et al, 2012). "
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    ABSTRACT: Group B streptococci (GBS) are Gram-positive bacteria that cause infections in utero and in newborns. We recently showed that the GBS pigment is hemolytic and increased pigment production promotes bacterial penetration of human placenta. However, mechanisms utilized by the hemolytic pigment to induce host cell lysis and the consequence on fetal injury are not known. Here, we show that the GBS pigment induces membrane permeability in artificial lipid bilayers and host cells. Membrane defects induced by the GBS pigment trigger K(+) efflux leading to osmotic lysis of red blood cells or pyroptosis in human macrophages. Macrophages lacking the NLRP3 inflammasome recovered from pigment-induced cell damage. In a murine model of in utero infection, hyperpigmented GBS strains induced fetal injury in both an NLRP3 inflammasome-dependent and NLRP3 inflammasome-independent manner. These results demonstrate that the dual mechanism of action of the bacterial pigment/lipid toxin leading to hemolysis or pyroptosis exacerbates fetal injury and suggest that preventing both activities of the hemolytic lipid is likely critical to reduce GBS fetal injury and preterm birth. © 2015 The Authors. Published under the terms of the CC BY 4.0 license.
    Full-text · Article · Mar 2015 · EMBO Molecular Medicine
    • "Screening of virulence factors in GBS vaginal clinical isolates Haemolysin extraction and quantification of haemolytic activity GBS haemolysin was extracted as described by Liu et al. (2004). The haemolytic titer was assessed by a microtiter dilution method, as described previously (Nizet et al. 1996). "
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    ABSTRACT: AimsTo assess the preventive effect of different intravaginal (i.va.) doses of Lactobacillus reuteri CRL1324 against vaginal colonization by Group B Streptococcus (GBS) in a murine experimental model.Methods and ResultsThe major virulence factors of four vaginal GBS clinical isolates were determined to select the most virulent strain and set up a murine model of streptococcal vaginal colonization. Later, the effect of four and seven doses of 108 viable cells of Lact. reuteri CRL1324 i.va. administered, prior to the GBS challenge was studied. Seven doses of lactobacilli were able to significantly reduce the number of viable GBS cells, while four doses showed no preventive effect. Both doses reduced the leukocyte influx induced by GBS. Seven doses caused a slight increase in the Lact. reuteri CRL1324 vaginal colonization compared with four doses and reduced murine vaginal pH compared to control mice.Conclusions Lact. reuteri CRL1324 evidenced a preventive effect on GBS vaginal colonization in an experimental mouse model.Significance and impacts of the studyMaternal GBS colonization is one of the most important risk factors for developing disease in newborns. Lact. reuteri CRL1324 could be considered as a new biological agent to reduce infections caused by this microorganism.This article is protected by copyright. All rights reserved.
    No preview · Article · Dec 2014 · Journal of Applied Microbiology
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