Acknowledgments We thank Alexander Johnson (yhb1D/D), Karl Kuchler (sodD/D mutants), Janet Quinn (hog1D/D, hog1/cap1D/D, trx1D/D) and Peter Staib (ssu1D/D) for providing mutant strains. We acknowledge helpful discussions with our colleagues from the Microbial Pathogenicity Mechanisms Department, Fungal Septomics and the Microbial Biochemistry and Physiology Research Group at the Hans Kno¨ll Institute (HKI), specially Ilse D. Jacobsen, Duncan Wilson, Sascha Brunke, Lydia Kasper, Franziska Gerwien, Sea´na Duggan, Katrin Haupt, Kerstin Hu¨nniger, and Matthias Brock, as well as from our partners in the FINSysB Network. Author Contributions Conceived and designed the experiments: PM HW IMB AJPB OK BH. Performed the experiments: PM CD HW. Analyzed the data: PM HW IMB AJPB OK BH. Wrote the paper: PM HW OK AJPB BH.Peer reviewedPublisher PD

A Cederlund

A da Silva Dantas

Alistair J. P. Brown

B Amulic

B Enjalbert

B Hube

BD Ullmann

Bernhard Hube

BH Youseff

BS Hromatka

C Fradin

Christine Dunker

CJ Barelle

CJ Heilmann

D Wilson

DA Smith

DL Moyes

DM Arana

DR Wysong

FC Fang

GM Chaves

Hanna Windecker

I Rubin-Bejerano

I Wozniok

ID Jacobsen

IE Frohner

Iryna M. Bohovych

J Cheetham

M de Jesus-Berrios

M Martchenko

MA Wheeler

MC Lorenz

MG Netea

Oliver Kurzai

Oscar Zaragoza

P Nordenfelt

Pedro Miramón

R Alonso-Monge

R Pathania

RA Calderone

RI Lehrer

SC Cheng

TM Stevanin

V Brinkmann

W Bellamy

W Chiranand

PLoS ONE

English

PubMed

Neutrophils are key players during Candida albicans infection. However, the relative contributions of neutrophil activities to fungal clearance and the relative importance of the fungal responses that counteract these activities remain unclear. We studied the contributions of the intra- and extracellular antifungal activities of human neutrophils using diagnostic Green Fluorescent Protein (GFP)-marked C. albicans strains. We found that a carbohydrate starvation response, as indicated by up-regulation of glyoxylate cycle genes, was only induced upon phagocytosis of the fungus. Similarly, the nitrosative stress response was only observed in internalised fungal cells. In contrast, the response to oxidative stress was observed in both phagocytosed and non-phagocytosed fungal cells, indicating that oxidative stress is imposed both intra- and extracellularly. We assessed the contributions of carbohydrate starvation, oxidative and nitrosative stress as antifungal activities by analysing the resistance to neutrophil killing of C. albicans mutants lacking key glyoxylate cycle, oxidative and nitrosative stress genes. We found that the glyoxylate cycle plays a crucial role in fungal resistance against neutrophils. The inability to respond to oxidative stress (in cells lacking superoxide dismutase 5 or glutathione reductase 2) renders C. albicans susceptible to neutrophil killing, due to the accumulation of reactive oxygen species (ROS). We also show that neutrophil

CiteSeerX

Cellular responses of Candida albicans to phagocytosis and the extracellular activities of neutrophils are critical to counteract carbohydrate starvation, oxidative and nitrosative stress. PLoS One 7: e52850

Acknowledgments We thank Alexander Johnson (yhb1D/D), Karl Kuchler (sodD/D mutants), Janet Quinn (hog1D/D, hog1/cap1D/D, trx1D/D) and Peter Staib (ssu1D/D) for providing mutant strains. We acknowledge helpful discussions with our colleagues from the Microbial Pathogenicity Mechanisms Department, Fungal Septomics and the Microbial Biochemistry and Physiology Research Group at the Hans Kno¨ll Institute (HKI), specially Ilse D. Jacobsen, Duncan Wilson, Sascha Brunke, Lydia Kasper, Franziska Gerwien, Sea´na Duggan, Katrin Haupt, Kerstin Hu¨nniger, and Matthias Brock, as well as from our partners in the FINSysB Network. Author Contributions Conceived and designed the experiments: PM HW IMB AJPB OK BH. Performed the experiments: PM CD HW. Analyzed the data: PM HW IMB AJPB OK BH. Wrote the paper: PM HW OK AJPB BH.Peer reviewe

Miramón, Pedro

Dunker, Christine

Windecker, Hanna

Bohovych, Iryna M

Brown, Alistair J P

Kurzai, Oliver

Hube, Bernhard

Aberdeen University Research

A biphasic innate immune MAPK response discriminates between the yeast and hyphal forms of Candida albicans in epithelial cells.

a c o b s e nI D ,W i l s o nD ,W a c h t l e rB ,B r u n k eS ,N a g l i kJ

A conserved stress-activated protein kinase regulates a core stress response in the human pathogen Candida albicans.

An integrated model of the recognition of Candida albicans by the innate immune system.

Antimicrobial polypeptides of human neutrophils.

Antimicrobial reactive oxygen and nitrogen species: concepts and controversies.

Bacterial infection induces nitric oxide synthase in human neutrophils.

C. albicans Cell Surface Superoxide Dismutases Degrade Host-Derived Reactive Oxygen Species to Escape Innate Immune Surveillance.

Candida albicans GRX2, encoding a putative glutaredoxin, is required for virulence in a murine model.

Candida albicans yeast and hyphae are discriminated by MAPK signaling in vaginal epithelial cells.

Candida and Candidosis.

Cloning and sequencing of a Candida albicans catalase gene and effects of disruption of this gene.

Combinatorial stresses kill pathogenic Candida species. Med Mycol.

CTA4 transcription factor mediates induction of nitrosative stress response in Candida albicans.

Differential susceptibility of mitogen-activated protein kinase pathway mutants to oxidativemediated killing by phagocytes in the fungal pathogen Candida albicans.

Enzymes that counteract nitrosative stress promote fungal virulence.

Extracellular superoxide dismutase protects Histoplasma yeast cells from host-derived oxidative stress.

Flavohemoglobin Hmp protects Salmonella enterica serovar typhimurium from nitric oxide-related killing by human macrophages.

From commensal to pathogen: stage- and tissue-specific gene expression of Candida albicans.

Fungal adaptation to the host environment.

GFP as a quantitative reporter of gene regulation in Candida albicans.

Granulocytes govern the transcriptional response, morphology and proliferation of Candida albicans in human blood.

Hyphal induction in the human fungal pathogen Candida albicans reveals a characteristic wall protein profile.

Identifying infection-associated genes of Candida albicans in the postgenomic era.

Inducible defense mechanism against nitric oxide in Candida albicans.

Induction of ERK-kinase signalling triggers morphotype-specific killing of Candida albicans filaments by human neutrophils.

Killing of Candida albicans by lactoferricin B, a potent antimicrobial peptide derived from the N-terminal region of bovine lactoferrin.

MAPKKK-independent regulation of the Hog1 stress-activated protein kinase in Candida albicans.

Neutrophil extracellular traps kill bacteria.

Niche-specific activation of the oxidative stress response by the pathogenic fungus Candida albicans.

Nichespecific regulation of central metabolic pathways in a fungal pathogen.

Nitric oxide scavenging and detoxification by the Mycobacterium tuberculosis haemoglobin, HbN in Escherichia coli.

Phagocytosis by neutrophils induces an amino acid deprivation response in Saccharomyces cerevisiae and Candida albicans.

Phagosome dynamics during phagocytosis by neutrophils.

Role of the Hog1 stress-activated protein kinase in the global transcriptional response to stress in the fungal pathogen Candida albicans.

Role of the mitogen-activated protein kinase Hog1p in morphogenesis and virulence of Candida albicans.

Specificity in killing pathogens is mediated by distinct repertoires of human neutrophil peptides.

Superoxide dismutases in Candida albicans: transcriptional regulation and functional characterization of the hyphal-induced SOD5 gene.

The dectin-1/inflammasome pathway is responsible for the induction of protective T-helper 17 responses that discriminate between yeasts and hyphae of Candida albicans.

The glyoxylate cycle is required for fungal virulence.

The hmp gene encoding the NOinducible flavohaemoglobin in Escherichia coli confers a protective advantage in resisting killing within macrophages, but not in vitro: links with swarming motility.

Thioredoxin regulates multiple hydrogen peroxide-induced signaling pathways in Candida albicans.

Transcriptional response of Candida albicans to nitric oxide and the role of the YHB1 gene in nitrosative stress and virulence.

Transcriptional response of Candida albicans upon internalization by macrophages.

Virulence factors of Candida albicans.

Zychlinsky A

Cellular responses of Candida albicans to phagocytosis and the extracellular activities of neutrophils are critical to counteract carbohydrate starvation, oxidative and nitrosative stress

Miramón Pedro

Dunker Christine

Windecker Hanna

Bohovych Iryna M.

Brown Alistair J. P.

Kurzai Oliver

Hube Bernhard

Public Library of Science (PLOS)

 to Phagocytosis and the Extracellular Activities of Neutrophils Are Critical to Counteract Carbohydrate Starvation, Oxidative and Nitrosative Stress

<div>Neutrophils are key players during Candida albicans infection. However, the relative contributions of neutrophil activities to fungal clearance and the relative importance of the fungal responses that counteract these activities remain unclear. We studied the contributions of the intra- and extracellular antifungal activities of human neutrophils using diagnostic Green Fluorescent Protein (GFP)-marked C. albicans strains. We found that a carbohydrate starvation response, as indicated by up-regulation of glyoxylate cycle genes, was only induced upon phagocytosis of the fungus. Similarly, the nitrosative stress response was only observed in internalised fungal cells. In contrast, the response to oxidative stress was observed in both phagocytosed and non-phagocytosed fungal cells, indicating that oxidative stress is imposed both intra- and extracellularly. We assessed the contributions of carbohydrate starvation, oxidative and nitrosative stress as antifungal activities by analysing the resistance to neutrophil killing of C. albicans mutants lacking key glyoxylate cycle, oxidative and nitrosative stress genes. We found that the glyoxylate cycle plays a crucial role in fungal resistance against neutrophils. The inability to respond to oxidative stress (in cells lacking superoxide dismutase 5 or glutathione reductase 2) renders C. albicans susceptible to neutrophil killing, due to the accumulation of reactive oxygen species (ROS). We also show that neutrophil-derived nitric oxide is crucial for the killing of C. albicans: a yhb1Δ/Δ mutant, unable to detoxify NO•, was more susceptible to neutrophils, and this phenotype was rescued by the nitric oxide scavenger carboxy-PTIO. The stress responses of C. albicans to neutrophils are partially regulated via the stress regulator Hog1 since a hog1Δ/Δ mutant was clearly less resistant to neutrophils and unable to respond properly to neutrophil-derived attack. Our data indicate that an appropriate fungal response to all three antifungal activities, carbohydrate starvation, nitrosative stress and oxidative stress, is essential for full wild type resistance to neutrophils. </div

Pedro Miramón (108557)

Christine Dunker (108560)

Hanna Windecker (108562)

Iryna M. Bohovych (108563)

Alistair J. P. Brown (108565)

Oliver Kurzai (108568)

Bernhard Hube (108571)

The Francis Crick Institute

Cellular Responses of Candida albicans to Phagocytosis and the Extracellular Activities of Neutrophils Are Critical to Counteract Carbohydrate Starvation, Oxidative and Nitrosative Stress

Neutrophils are key players during Candida albicans infection. However, the relative contributions of neutrophil activities to fungal clearance and the relative importance of the fungal responses that counteract these activities remain unclear. We studied the contributions of the intra- and extracellular antifungal activities of human neutrophils using diagnostic Green Fluorescent Protein (GFP)-marked C. albicans strains. We found that a carbohydrate starvation response, as indicated by up-regulation of glyoxylate cycle genes, was only induced upon phagocytosis of the fungus. Similarly, the nitrosative stress response was only observed in internalised fungal cells. In contrast, the response to oxidative stress was observed in both phagocytosed and non-phagocytosed fungal cells, indicating that oxidative stress is imposed both intra- and extracellularly. We assessed the contributions of carbohydrate starvation, oxidative and nitrosative stress as antifungal activities by analysing the resistance to neutrophil killing of C. albicans mutants lacking key glyoxylate cycle, oxidative and nitrosative stress genes. We found that the glyoxylate cycle plays a crucial role in fungal resistance against neutrophils. The inability to respond to oxidative stress (in cells lacking superoxide dismutase 5 or glutathione reductase 2) renders C. albicans susceptible to neutrophil killing, due to the accumulation of reactive oxygen species (ROS). We also show that neutrophil-derived nitric oxide is crucial for the killing of C. albicans: a yhb1Δ/Δ mutant, unable to detoxify NO•, was more susceptible to neutrophils, and this phenotype was rescued by the nitric oxide scavenger carboxy-PTIO. The stress responses of C. albicans to neutrophils are partially regulated via the stress regulator Hog1 since a hog1Δ/Δ mutant was clearly less resistant to neutrophils and unable to respond properly to neutrophil-derived attack. Our data indicate that an appropriate fungal response to all three antifungal activities, carbohydrate starvation, nitrosative stress and oxidative stress, is essential for full wild type resistance to neutrophils

Iryna M Bohovych

Alistair J P Brown

Directory of Open Access Journals

Cellular responses of Candida albicans to phagocytosis and the extracellular activities of neutrophils are critical to counteract carbohydrate starvation, oxidative and nitrosative stress.

Crossref

Cellular Responses of Candida albicans to Phagocytosis and the Extracellular Activities of Neutrophils Are Critical to Counteract Carbohydrate Starvation, Oxidative and Nitrosative Stress

https://figshare.com/articles/Cellular_Responses_of_Candida_albicans_to_Phagocytosis_and_the_Extracellular_Activities_of_Neutrophils_Are_Critical_to_Counteract_Carbohydrate_Starvation_Oxidative_and_Nitrosative_Stress__/115740

Cellular responses of Candida albicans to phagocytosis and the extracellular activities of neutrophils are critical to counteract carbohydrate starvation, oxidative and nitrosative stress

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