Streptococcus pyogenes bacteria modulate membrane traffic in human neutrophils and selectively inhibit azurophilic granule fusion with phagosomes

Group A Streptococcus pyogenes (GAS) strain AP53 is a primary isolate from a patient with necrotizing fasciitis. These AP53 cells contain an inactivating mutation in the sensor component of the cluster of virulence (cov) responder (R)/sensor (S) two-component gene regulatory system (covRS), which enhances the virulence of the primary strain, AP53/covR+S−. However, specific mechanisms by which the covRS system regulates the survival of GAS in humans are incomplete. Here, we show a key role for covRS in the regulation of opsonophagocytosis of AP53 by human neutrophils. AP53/covR+S− cells displayed potent binding of host complement inhibitors of C3 convertase, viz. Factor H (FH) and C4-binding protein (C4BP), which concomitantly led to minimal C3b deposition on AP53 cells, further showing that these plasma protein inhibitors are active on GAS cells. This resulted in weak killing of the bacteria by human neutrophils and a corresponding high death rate of mice after injection of these cells. After targeted allelic alteration of covS− to wild-type covS (covS+), a dramatic loss of FH and C4BP binding to the AP53/covR+S+ cells was observed. This resulted in elevated C3b deposition on AP53/covR+S+ cells, a high level of opsonophagocytosis by human neutrophils, and a very low death rate of mice infected with AP53/covR+S+. We show that covRS is a critical transcriptional regulator of genes directing AP53 killing by neutrophils and regulates the levels of the receptors for FH and C4BP, which we identify as the products of the fba and enn genes, respectively.

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Streptococcus pyogenes and human neutrophils: a paradigm for evasion of innate host defense by bacterial pathogens

Microbes and Infection ◽

10.1016/j.micinf.2004.05.022 ◽

2004 ◽

Vol 6 (12) ◽

pp. 1117-1123 ◽

Cited By ~ 42

Author(s):

Jovanka M. Voyich ◽

James M. Musser ◽

Frank R. DeLeo

Keyword(s):

Streptococcus Pyogenes ◽

Host Defense ◽

Bacterial Pathogens ◽

Human Neutrophils

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LXA4, aspirin-triggered 15-epi-LXA4, and their analogs selectively downregulate PMN azurophilic degranulation

AJP Cell Physiology ◽

10.1152/ajpcell.1999.276.4.c988 ◽

1999 ◽

Vol 276 (4) ◽

pp. C988-C994 ◽

Cited By ~ 45

Author(s):

Andrew T. Gewirtz ◽

Valery V. Fokin ◽

Nicos A. Petasis ◽

Charles N. Serhan ◽

James L. Madara

Keyword(s):

Inflammatory Disease ◽

Inflammatory Mediator ◽

Lipid Mediators ◽

Human Neutrophils ◽

Causative Role ◽

Cellular Mechanisms ◽

Azurophilic Granule ◽

Disease States ◽

Granule Release

The eicosanoid lipoxin A4(LXA4) is biosynthesized in vivo by cells present at inflammatory sites and appears to be an endogenous anti-inflammatory mediator. Further, in the presence of aspirin, the 15-epimer of LXA4(15-epi-LXA4) is biosynthesized and may mediate some of aspirin’s desirable bioactions. LXA4, 15-epi-LXA4, and their stable analogs inhibit inflammation in established animal models, indicating that these compounds may be useful for treating inflammatory disease states. To investigate the cellular mechanisms by which these lipid mediators downregulate inflammation, we investigated whether these eicosanoids could influence receptor-mediated degranulation of human neutrophils, an event thought to play a major causative role in several inflammatory disease states. LXA4, 15-epi-LXA4, and their stable analogs potently (IC50< 1 nM) and selectively downregulated neutrophil release of azurophilic granule contents but did not affect other neutrophil secretory functions. Thus the cellular basis of action of these natural off-switches to inflammation appears to involve downregulation of neutrophil azurophilic granule release.

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A requirement for neutrophil glycosaminoglycans in chemokine:receptor interactions is revealed by the streptococcal protease SpyCEP

10.1101/461418 ◽

2018 ◽

Author(s):

Jenny Goldblatt ◽

Richard Ashley Lawrenson ◽

Luke Muir ◽

Saloni Dattani ◽

Tomoko Tsuchiya ◽

...

Keyword(s):

Streptococcus Pyogenes ◽

Chemokine Receptors ◽

Human Neutrophils ◽

Invasive Infection ◽

Human Pathogen ◽

Core Domain ◽

The Core ◽

Disulphide Bonds ◽

Impaired Binding

To evade the immune system, the lethal human pathogenStreptococcus pyogenesproduces SpyCEP, an enzyme that cleaves the C-terminalα-helix of CXCL8, resulting in markedly impaired recruitment of neutrophils to sites of invasive infection. The basis for chemokine inactivation by SpyCEP is, however, poorly understood, as the core domain of CXCL8 known to interact with CXCL8 receptors is unaffected by enzymatic cleavage.We examined thein vitromigration of human neutrophils and observed that their ability to efficiently navigate a CXCL8 gradient was compromised following CXCL8 cleavage by SpyCEP. SpyCEP-mediated cleavage of CXCL8 also impaired CXCL8-induced migration of transfectants expressing the human chemokine receptors CXCR1 or CXCR2. Despite possessing an intact N-terminus and preserved disulphide bonds, SpyCEP-cleaved CXCL8 had impaired binding to both CXCR1 and CXCR2, pointing to a requirement for the C terminalα-helix. SpyCEP-cleaved CXCL8 had similarly impaired binding to the glycosaminoglycan heparin. Enzymatic removal of neutrophil glycosaminoglycans was observed to ablate neutrophil navigation of a CXCL8 gradient, whilst navigation of an fMLP gradient remained largely intact.We conclude therefore, that SpyCEP cleavage of CXCL8 results in chemokine inactivation due to a requirement for glycosaminoglycan binding in productive chemokine:receptor interactions. This may inform strategies to inhibit the activity of SpyCEP, but may also influence future approaches to inhibit unwanted chemokine-induced inflammation.

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Streptococcus pyogenes expressing M and M-like surface proteins are phagocytosed but survive inside human neutrophils

Cellular Microbiology ◽

10.1046/j.1462-5822.2003.00272.x ◽

2003 ◽

Vol 5 (4) ◽

pp. 253-265 ◽

Cited By ~ 57

Author(s):

Leila Staali ◽

Matthias Morgelin ◽

Lars Bjorck ◽

Hans Tapper

Keyword(s):

Streptococcus Pyogenes ◽

Surface Proteins ◽

Human Neutrophils

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Neutrophil Myeloperoxidase Destruction by Ultraviolet Irradiation

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100102778 ◽

1988 ◽

Vol 46 ◽

pp. 138-139

Author(s):

J. Hanker ◽

B. Giammara ◽

G. Strauss

Keyword(s):

Uv Radiation ◽

Myeloid Leukemia ◽

Stratospheric Ozone ◽

Psoriasis Patient ◽

Whole Body ◽

Human Neutrophils ◽

Cupric Nitrate ◽

The Earth ◽

Routine Handling ◽

Uvb Phototherapy

Only a fraction of the UV radiation emitted by the sun reaches the earth; most of the UVB (290-320nm) is eliminated by stratospheric ozone. There is increasing concern, however, that man-made chemicals are damaging this ozone layer. Although the effects of UV on DNA or as a carcinogen are widely known, preleukemia and acute myeloid leukemia (AML) have only rarely been reported in psoriasis patients treated with 8-methoxypsoralen and UV (PUVA). It was therefore of interest to study the effects of UV on the myeloperoxidase (MP) activity of human neutrophils. The peroxidase activity of enriched leukocyte preparations on coverslips was shown cytochemically with a diaminobenzidine medium and cupric nitrate intensification.Control samples (Figs. 1,4,5) of human bloods that were not specifically exposed to UV radiation or light except during routine handling were compared with samples which had been exposed in one of several different ways. One preparation (Fig. 2) was from a psoriasis patient who had received whole-body UVB phototherapy repeatedly.

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Application of high-resolution field-emission LVSEM, backscatter imaging, immunogold staining to definition of the spatial distributions of two human neutrophil adherence receptors

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100146023 ◽

1993 ◽

Vol 51 ◽

pp. 36-37

Author(s):

Robert D. Nelson ◽

Sharon R. Hasslen ◽

Stanley L. Erlandsen

Keyword(s):

Cell Surface ◽

Surface Membrane ◽

Three Dimensional ◽

Human Neutrophils ◽

Spatial Distributions ◽

Immunogold Staining ◽

Functional Control ◽

Neutrophil Adherence ◽

Definition Of ◽

Mode Of Attachment

Receptors are commonly defined in terms of number per cell, affinity for ligand, chemical structure, mode of attachment to the cell surface, and mechanism of signal transduction. We propose to show that knowledge of spatial distribution of receptors on the cell surface can provide additional clues to their function and components of functional control.L-selectin and Mac-1 denote two receptor populations on the neutrophil surface that mediate neutrophil-endothelial cell adherence interactions and provide for targeting of neutrophil recruitment to sites of inflammation. We have studied the spatial distributions of these receptors using LVSEM and backscatter imaging of isolated human neutrophils stained with mouse anti-receptor (primary) antibody and goat anti-mouse (secondary) antibody conjugated to 12 nm colloidal gold. This combination of techniques provides for three-dimensional analysis of the expression of these receptors on different surface membrane domains of the neutrophil: the ruffles and microvilli that project from the cell surface, and the cell body between these projecting structures.

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