PUBLICATION

Neutrophils mediate Salmonella Typhimurium clearance through the GBP4 inflammasome-dependent production of prostaglandins

Authors

Tyrkalska, S.D., Candel, S., Angosto, D., Gómez-Abellán, V., Martín-Sánchez, F., García-Moreno, D., Zapata-Pérez, R., Sánchez-Ferrer, Á., Sepulcre, M.P., Pelegrín, P., Mulero, V.

ID

ZDB-PUB-160702-7

Date

2016

Source

Nature communications 7: 12077 (Journal)

Registered Authors

Mulero, Victor

Keywords

Bacterial infection, Inflammasome, Medical research, Neutrophils

MeSH Terms

Morpholinos
Prostaglandin D2/biosynthesis*
GTP-Binding Proteins/immunology*
Animals
Inflammasomes/immunology*
Zebrafish
Leukotriene B4/immunology
Salmonella typhimurium
Organisms, Genetically Modified
Neutrophils/immunology*
Caspase Activation and Recruitment Domain
Prostaglandins/immunology
Interleukin-8/immunology

(all 13)

PubMed

27363812 Full text @ Nat. Commun.

Abstract

Inflammasomes are cytosolic molecular platforms that alert the immune system about the presence of infection. Here we report that zebrafish guanylate-binding protein 4 (Gbp4), an IFNγ-inducible GTPase protein harbouring a C-terminal CARD domain, is required for the inflammasome-dependent clearance of Salmonella Typhimurium (ST) by neutrophils in vivo. Despite the presence of the CARD domain, Gbp4 requires the universal inflammasome adaptor Asc for mediating its antibacterial function. In addition, the GTPase activity of Gbp4 is indispensable for inflammasome activation and ST clearance. Mechanistically, neutrophils are recruited to the infection site through the inflammasome-independent production of the chemokine (CXC motif) ligand 8 and leukotriene B4, and then mediate bacterial clearance through the Gbp4 inflammasome-dependent biosynthesis of prostaglandin D2. Our results point to GBPs as key inflammasome adaptors required for prostaglandin biosynthesis and bacterial clearance by neutrophils and suggest that transient activation of the inflammasome may be used to treat bacterial infections.

Genes / Markers

Marker	Marker Type	Name
alox5b.2	GENE	arachidonate 5-lipoxygenase b, tandem duplicate 2
alox5b.3	GENE	arachidonate 5-lipoxygenase b, tandem duplicate 3
caspa	GENE	caspase a
caspb	GENE	caspase b
csf3a	GENE	colony stimulating factor 3 (granulocyte) a
cxcr2	GENE	chemokine (C-X-C motif) receptor 2
gbp3	GENE	guanylate binding protein 3
gbp4	GENE	guanylate binding protein 4
il1b	GENE	interleukin 1, beta
lta4h	GENE	leukotriene A4 hydrolase

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Figures

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Expression

Gene	Fish	Conditions	Stage	Qualifier	Anatomy	Assay	Figure
EGFP	gl22Tg	bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium	Protruding-mouth		macrophage	RTPCR	Fig. 1
EGFP	gl22Tg	control	Protruding-mouth		macrophage	RTPCR	Fig. 1
EGFP	gl22Tg	bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium	Protruding-mouth		whole organism	RTPCR	Fig. 1
EGFP	gl22Tg	control	Protruding-mouth		whole organism	RTPCR	Fig. 1
EGFP	i114Tg	control	Protruding-mouth		neutrophil	IFL	Fig. S8
EGFP	i114Tg + MO1-gbp4	control	Protruding-mouth		neutrophil	IFL	Fig. S8
gbp4	gl22Tg	bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium	Protruding-mouth	Not Detected	macrophage	RTPCR	Fig. 1
gbp4	gl22Tg	control	Protruding-mouth	Not Detected	macrophage	RTPCR	Fig. 1
gbp4	gl22Tg	control	Protruding-mouth		whole organism	RTPCR	Fig. 1
gbp4	gl22Tg	bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium	Protruding-mouth		whole organism	RTPCR	Fig. 1

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT	chemical treatment by environment: prostaglandin J2, bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium	Long-pec to Days 7-13	whole organism decreased life span, abnormal	Fig. 9
WT	chemical treatment by environment: prostaglandin J2, bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium	Protruding-mouth	cysteine-type endopeptidase activity increased occurrence, abnormal	Fig. 9
WT	bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium, chemical treatment by environment: 15-deoxy-Delta(12,14)-prostaglandin J2	Long-pec to Days 7-13	whole organism decreased life span, exacerbated	Fig. 9
WT	bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium, chemical treatment by environment: 15-deoxy-Delta(12,14)-prostaglandin J2	Protruding-mouth	cysteine-type endopeptidase activity decreased occurrence, abnormal	Fig. 9
WT	chemical treatment by environment: N-(p-amylcinnamoyl)anthranilic acid, bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium	Long-pec to Days 7-13	whole organism decreased life span, exacerbated	Fig. 8
WT	bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium, chemical treatment by environment: indometacin	Long-pec to Days 7-13	whole organism decreased life span, abnormal	Fig. 8
WT	chemical treatment by environment: prostaglandin D2, bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium	Long-pec to Days 7-13	whole organism decreased life span, abnormal	Fig. 9
WT	chemical treatment by environment: prostaglandin D2, bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium	Protruding-mouth	cysteine-type endopeptidase activity increased occurrence, abnormal	Fig. 9
WT	bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium, chemical treatment by environment: prostaglandin E2	Long-pec to Days 7-13	whole organism decreased life span, abnormal	Fig. 9
WT	bacterial treatment by injection: Salmonella enterica subsp. enterica serovar Typhimurium, chemical treatment by environment: prostaglandin E2	Protruding-mouth	cysteine-type endopeptidase activity increased occurrence, abnormal	Fig. 9

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Mutations / Transgenics

Allele	Construct	Type
gl22Tg	Tg(mpeg1:EGFP)	Transgenic Insertion
i114Tg	Tg(mpx:GFP)	Transgenic Insertion
nz50Tg	Tg(lyz:DsRed2)	Transgenic Insertion
sh267Tg	TgBAC(mpx:GAL4-VP16)	Transgenic Insertion
ums2Tg	Tg(UAS:gbp4,myl7:EGFP)	Transgenic Insertion
ums3Tg	Tg(UAS:dngbp4,myl7:EGFP)	Transgenic Insertion

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
cxcr2	MO1-cxcr2	MRPHLNO
gbp4	MO1-gbp4	MRPHLNO
il1b	MO1-il1b	MRPHLNO
lta4h	MO1-lta4h	MRPHLNO
pycard	MO3-pycard	MRPHLNO
pycard	MO4-pycard	MRPHLNO

Fish

Fish
gl22Tg
i114Tg
i114Tg + MO1-gbp4
sh267Tg; ums2Tg
sh267Tg; ums2Tg + MO1-gbp4
sh267Tg; ums3Tg
WT
WT + MO1-cxcr2
WT + MO1-gbp4
WT + MO1-lta4h

Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
DsRed2	EFG	DsRed2
EGFP	EFG	EGFP
GAL4	EFG	GAL4
GFP	EFG	GFP

Mapping

Name	Type	Antigen Genes	Isotypes	Host Organism
Ab1-gbp4		gbp4
Ab2-gbp4		gbp4
Ab3-gbp4		gbp4
Ab4-gbp4		gbp4

Tyrkalska et al., 2016 ZDB-PUB-160702-7 PMID:27363812

Neutrophils mediate Salmonella Typhimurium clearance through the GBP4 inflammasome-dependent production of prostaglandins

Tyrkalska et al., 2016

ZDB-PUB-160702-7
PMID:27363812