New Insights into Neutrophil Extracellular Trap (NETs) Formation from Porcine Neutrophils in Response to Bacterial Infections

Actinobacillus pleuropneumoniae (A.pp, Gram negative) and Streptococcus (S.) suis (Gram positive) can cause severe diseases in pigs. During infection, neutrophils infiltrate to counteract these pathogens with phagocytosis and/or neutrophil extracellular traps (NETs). NETs consist of a DNA-backbone s...

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Published in:International journal of molecular sciences 2022-08, Vol.23 (16), p.8953
Main Authors: Bonilla, Marta C., Quiros, Oriana N., Wendt, Michael, Hennig-Pauka, Isabel, Mörgelin, Matthias, von Köckritz-Blickwede, Maren, de Buhr, Nicole
Format: Article
Language:English
Subjects:
Antiinfectives and antibacterials
Bacteria
Bacterial infections
CD18 antigen
Cytokines
Enzymes
Infections
Leukocytes (neutrophilic)
Markers
Microscopy
Mitochondrial DNA
NAD(P)H oxidase
Neutrophils
Pathogens
Phagocytosis
Phagolysosomes
Phenotypes
Pneumonia
Streptococcus infections
Swine
Transmission electron microscopy
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Summary:Actinobacillus pleuropneumoniae (A.pp, Gram negative) and Streptococcus (S.) suis (Gram positive) can cause severe diseases in pigs. During infection, neutrophils infiltrate to counteract these pathogens with phagocytosis and/or neutrophil extracellular traps (NETs). NETs consist of a DNA-backbone spiked with antimicrobial components. The NET formation mechanisms in porcine neutrophils as a response to both of the pathogens are not entirely clear. The aim of this study was to investigate whether A.pp (serotype 2, C3656/0271/11) and S. suis (serotype 2, strain 10) induce NETs by NADPH oxidase- or CD18-dependent mechanisms and to characterize phenotypes of NETs in porcine neutrophils. Therefore, we investigated NET induction in porcine neutrophils in the presence and absence of NET inhibitors and quantified NETs after 3 h. Furthermore, NETosis and phagocytosis were investigated by transmission electron microscopy after 30 min to characterize different phenotypes. A.pp and S. suis induce NETs that are mainly ROS-dependent. A.pp induces NETs that are partially CD18-dependent. Thirty minutes after infection, both of the pathogens induced a vesicular NET formation with only slight differences. Interestingly, some neutrophils showed only NET-marker positive phagolysosomes, but no NET-marker positive vesicles. Other neutrophils showed vesicular NETs and only NET-marker negative phagolysosomes. In conclusion, both of the pathogens induce ROS-dependent NETs. Vesicular NETosis and phagocytosis occur in parallel in porcine neutrophils in response to S. suis serotype 2 and A.pp serotype 2.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms23168953