Neutrophil-derived reactive agents induce a transient SpeB negative phenotype in Streptococcus pyogenes.
Necrotizing soft tissue infections
Neutrophils
SpeB
Streptococcus pyogenes
Journal
Journal of biomedical science
ISSN: 1423-0127
Titre abrégé: J Biomed Sci
Pays: England
ID NLM: 9421567
Informations de publication
Date de publication:
10 Jul 2023
10 Jul 2023
Historique:
received:
24
02
2023
accepted:
03
07
2023
medline:
12
7
2023
pubmed:
11
7
2023
entrez:
10
7
2023
Statut:
epublish
Résumé
Streptococcus pyogenes (group A streptococci; GAS) is the main causative pathogen of monomicrobial necrotizing soft tissue infections (NSTIs). To resist immuno-clearance, GAS adapt their genetic information and/or phenotype to the surrounding environment. Hyper-virulent streptococcal pyrogenic exotoxin B (SpeB) negative variants caused by covRS mutations are enriched during infection. A key driving force for this process is the bacterial Sda1 DNase. Bacterial infiltration, immune cell influx, tissue necrosis and inflammation in patient´s biopsies were determined using immunohistochemistry. SpeB secretion and activity by GAS post infections or challenges with reactive agents were determined via Western blot or casein agar and proteolytic activity assays, respectively. Proteome of GAS single colonies and neutrophil secretome were profiled, using mass spectrometry. Here, we identify another strategy resulting in SpeB-negative variants, namely reversible abrogation of SpeB secretion triggered by neutrophil effector molecules. Analysis of NSTI patient tissue biopsies revealed that tissue inflammation, neutrophil influx, and degranulation positively correlate with increasing frequency of SpeB-negative GAS clones. Using single colony proteomics, we show that GAS isolated directly from tissue express but do not secrete SpeB. Once the tissue pressure is lifted, GAS regain SpeB secreting function. Neutrophils were identified as the main immune cells responsible for the observed phenotype. Subsequent analyses identified hydrogen peroxide and hypochlorous acid as reactive agents driving this phenotypic GAS adaptation to the tissue environment. SpeB-negative GAS show improved survival within neutrophils and induce increased degranulation. Our findings provide new information about GAS fitness and heterogeneity in the soft tissue milieu and provide new potential targets for therapeutic intervention in NSTIs.
Sections du résumé
BACKGROUND
BACKGROUND
Streptococcus pyogenes (group A streptococci; GAS) is the main causative pathogen of monomicrobial necrotizing soft tissue infections (NSTIs). To resist immuno-clearance, GAS adapt their genetic information and/or phenotype to the surrounding environment. Hyper-virulent streptococcal pyrogenic exotoxin B (SpeB) negative variants caused by covRS mutations are enriched during infection. A key driving force for this process is the bacterial Sda1 DNase.
METHODS
METHODS
Bacterial infiltration, immune cell influx, tissue necrosis and inflammation in patient´s biopsies were determined using immunohistochemistry. SpeB secretion and activity by GAS post infections or challenges with reactive agents were determined via Western blot or casein agar and proteolytic activity assays, respectively. Proteome of GAS single colonies and neutrophil secretome were profiled, using mass spectrometry.
RESULTS
RESULTS
Here, we identify another strategy resulting in SpeB-negative variants, namely reversible abrogation of SpeB secretion triggered by neutrophil effector molecules. Analysis of NSTI patient tissue biopsies revealed that tissue inflammation, neutrophil influx, and degranulation positively correlate with increasing frequency of SpeB-negative GAS clones. Using single colony proteomics, we show that GAS isolated directly from tissue express but do not secrete SpeB. Once the tissue pressure is lifted, GAS regain SpeB secreting function. Neutrophils were identified as the main immune cells responsible for the observed phenotype. Subsequent analyses identified hydrogen peroxide and hypochlorous acid as reactive agents driving this phenotypic GAS adaptation to the tissue environment. SpeB-negative GAS show improved survival within neutrophils and induce increased degranulation.
CONCLUSIONS
CONCLUSIONS
Our findings provide new information about GAS fitness and heterogeneity in the soft tissue milieu and provide new potential targets for therapeutic intervention in NSTIs.
Identifiants
pubmed: 37430325
doi: 10.1186/s12929-023-00947-x
pii: 10.1186/s12929-023-00947-x
pmc: PMC10331992
doi:
Substances chimiques
erythrogenic toxin
0
Bacterial Proteins
0
Exotoxins
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
52Subventions
Organisme : Deutsche Forschungsgemeinschaft
ID : 407176682
Organisme : Deutsche Forschungsgemeinschaft
ID : 492903360
Organisme : Deutsche Forschungsgemeinschaft
ID : 503880638
Organisme : Center for Innovative Medicine
ID : 20180058
Organisme : Vetenskapsrådet
ID : 2018-02475
Organisme : Vetenskapsrådet
ID : 2018-151
Organisme : VINNOVA
ID : 90456
Investigateurs
Morten Hedetoft
(M)
Trond Bruun
(T)
Oddvar Oppegaard
(O)
Torbjørn Nedrebø
(T)
Eivind Rath
(E)
Martin Bruun Madsen
(MB)
Informations de copyright
© 2023. The Author(s).
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