PUBLICATION

Mycobacterium abscessus-Induced Granuloma Formation Is Strictly Dependent on TNF Signaling and Neutrophil Trafficking

Authors

Bernut, A., Nguyen-Chi, M., Halloum, I., Herrmann, J.L., Lutfalla, G., Kremer, L.

ID

ZDB-PUB-161103-3

Date

2016

Source

PLoS pathogens 12: e1005986 (Journal)

Registered Authors

Lutfalla, Georges

Keywords

Neutrophils, Embryos, Macrophages, Granulomas, Larvae, Fluorescence microscopy, Zebrafish, Confocal microscopy

MeSH Terms

Animals
Animals, Genetically Modified
Signal Transduction/immunology*
Mycobacterium Infections, Nontuberculous/immunology*
Mycobacterium Infections, Nontuberculous/microbiology
Mycobacterium Infections, Nontuberculous/pathology
Chemotaxis, Leukocyte/immunology
Disease Models, Animal
Tumor Necrosis Factor-alpha/immunology*
Granuloma/immunology*
Granuloma/pathology
Microscopy
Zebrafish
Nontuberculous Mycobacteria/immunology
Polymerase Chain Reaction
Neutrophils/cytology
Neutrophils/immunology*

(all 17)

PubMed

27806130 Full text @ PLoS Pathog.

Abstract

Mycobacterium abscessus is considered the most common respiratory pathogen among the rapidly growing non-tuberculous mycobacteria. Infections with M. abscessus are increasingly found in patients with chronic lung diseases, especially cystic fibrosis, and are often refractory to antibiotic therapy. M. abscessus has two morphotypes with distinct effects on host cells and biological responses. The smooth (S) variant is recognized as the initial airway colonizer while the rough (R) is known to be a potent inflammatory inducer associated with invasive disease, but the underlying immunopathological mechanisms of the infection remain unsolved. We conducted a comparative stepwise dissection of the inflammatory response in S and R pathogenesis by monitoring infected transparent zebrafish embryos. Loss of TNFR1 function resulted in increased mortality with both variants, and was associated with unrestricted intramacrophage bacterial growth and decreased bactericidal activity. The use of transgenic zebrafish lines harboring fluorescent macrophages and neutrophils revealed that neutrophils, like macrophages, interact with M. abscessus at the initial infection sites. Impaired TNF signaling disrupted the IL8-dependent neutrophil mobilization, and the defect in neutrophil trafficking led to the formation of aberrant granulomas, extensive mycobacterial cording, unrestricted bacterial growth and subsequent larval death. Our findings emphasize the central role of neutrophils for the establishment and maintenance of the protective M. abscessus granulomas. These results also suggest that the TNF/IL8 inflammatory axis is necessary for protective immunity against M. abscessus and may be of clinical relevance to explain why immunosuppressive TNF therapy leads to the exacerbation of M. abscessus infections.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Qualifier	Anatomy	Assay	Figure
cxcl8a	WT	bacterial treatment by injection: Mycobacteroides abscessus	Day 5		whole organism	RTPCR	Fig. 5
cxcl8a	WT	bacterial treatment by injection: Mycobacteroides abscessus	Days 7-13		whole organism	RTPCR	Fig. 5
cxcl8a	WT	bacterial treatment by injection: Mycobacteroides abscessus	Day 5		whole organism	RTPCR	Fig. 8
cxcl8a	WT + MO3-tnfrsf1a	bacterial treatment by injection: Mycobacteroides abscessus	Day 5		whole organism	RTPCR	Fig. 8
DsRed2	nz50Tg; ump5Tg	bacterial treatment by injection: Mycobacteroides abscessus	Day 6		neutrophil	IFL	Fig. 1
EGFP	nz50Tg; ump5Tg	bacterial treatment by injection: Mycobacteroides abscessus	Day 6	Not Detected	neutrophil	IFL	Fig. 1
EGFP	ump2Tg; ump5Tg	bacterial treatment by injection: Mycobacteroides abscessus	Day 6		macrophage	IFL	Fig. 1
EGFP	ump5Tg	bacterial treatment by injection: Mycobacteroides abscessus	Day 6		anatomical structure	IFL	Fig. 1
EGFP	ump5Tg	bacterial treatment by injection: Mycobacteroides abscessus	Day 5		anatomical structure	IFL	Fig. 1
EGFP	ump5Tg	bacterial treatment by injection: Mycobacteroides abscessus	Days 7-13		anatomical structure	IFL	Fig. 1

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT	bacterial treatment by injection: Mycobacteroides abscessus	Protruding-mouth to Days 7-13	whole organism decreased life span, abnormal	Fig. 5
WT	bacterial treatment by injection: Mycobacteroides abscessus	Day 4 to Days 14-20	whole organism decreased life span, abnormal	Fig. 2
WT	bacterial treatment by injection: Mycobacteroides abscessus	Day 5	whole organism cxcl8a expression increased amount, abnormal	Fig. 8
WT	bacterial treatment by injection: Mycobacteroides abscessus	Day 5	whole organism cxcl8a expression increased amount, abnormal	Fig. 5
WT	bacterial treatment by injection: Mycobacteroides abscessus	Day 5	whole organism tnfa expression increased amount, abnormal	Fig. 1
WT	bacterial treatment by injection: Mycobacteroides abscessus	Days 7-13	whole organism cxcl8a expression increased amount, abnormal	Fig. 5
WT	bacterial treatment by injection: Mycobacteroides abscessus	Days 7-13	whole organism tnfa expression increased amount, abnormal	Fig. 1
WT + MO3-csf3r	bacterial treatment by injection: Mycobacteroides abscessus	Protruding-mouth to Days 7-13	whole organism decreased life span, abnormal	Fig. 5
WT + MO3-csf3r	bacterial treatment by injection: Mycobacteroides abscessus	Day 5	granuloma formation decreased occurrence, abnormal	Fig. 8
WT + MO3-csf3r	bacterial treatment by injection: Mycobacteroides abscessus	Day 5	response to bacterium decreased process quality, abnormal	Fig. 5

1 - 10 of 49

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Mutations / Transgenics

Allele	Construct	Type
i114Tg	Tg(mpx:GFP)	Transgenic Insertion
nz50Tg	Tg(lyz:DsRed2)	Transgenic Insertion
ump2Tg	Tg(mpeg1.1:mCherryF)	Transgenic Insertion
ump5Tg	Tg(tnfa:EGFPF)	Transgenic Insertion

1 - 4 of 4

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Human Disease / Model

Human Disease	Fish	Conditions	Evidence
bacterial infectious disease			TAS

Sequence Targeting Reagents

Target	Reagent	Reagent Type
csf3r	MO3-csf3r	MRPHLNO
cxcl8a	MO4-cxcl8a	MRPHLNO
tnfrsf1a	MO3-tnfrsf1a	MRPHLNO

Fish

Fish
i114Tg
i114Tg + MO3-tnfrsf1a
i114Tg + MO4-cxcl8a
nz50Tg; ump5Tg
ump2Tg
ump2Tg + MO3-tnfrsf1a
ump2Tg; ump5Tg
ump5Tg
WT
WT + MO3-csf3r

Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
DsRed2	EFG	DsRed2
EGFP	EFG	EGFP
GFP	EFG	GFP
mCherry	EFG	mCherry

Mapping

Bernut et al., 2016 ZDB-PUB-161103-3 PMID:27806130

Mycobacterium abscessus-Induced Granuloma Formation Is Strictly Dependent on TNF Signaling and Neutrophil Trafficking

Bernut et al., 2016

ZDB-PUB-161103-3
PMID:27806130