Relative contribution of Prevotella intermedia and Pseudomonas aeruginosa to lung pathology in airways of patients with cystic fibrosis

Martina Ulrich; Isabelle Beer; Peter Braitmaier; Michaela Dierkes; Florian Kummer; Bernhard Krismer; Ulrike Schumacher; Ute Gräpler-Mainka; Joachim Riethmüller; Peter Østrup Jensen; Thomas Bjarnsholt; Niels Høiby; Gabriel Bellon; Gerd Döring

doi:10.1136/thx.2010.137745

Relative contribution of Prevotella intermedia and Pseudomonas aeruginosa to lung pathology in airways of patients with cystic fibrosis

Martina Ulrich, Isabelle Beer, Peter Braitmaier, Michaela Dierkes, Florian Kummer, Bernhard Krismer, Ulrike Schumacher, Ute Gräpler-Mainka, Joachim Riethmüller, Peter Østrup Jensen, Thomas Bjarnsholt, Niels Høiby, Gabriel Bellon, Gerd Döring

LUKKET: Institut for Immunologi og Mikrobiologi

60 Citationer (Scopus)

Abstract

Background: Patients with cystic fibrosis (CF) with Pseudomonas aeruginosa lung infections produce endobronchial mucus plugs allowing growth of obligate anaerobes including Prevotella spp. Whether obligate anaerobes contribute to the pathophysiology of CF lung disease is unknown. Methods: The virulence of Prevotella intermedia and Ps aeruginosa was investigated in vitro and in mice, antibodies against P intermedia in CF sera were assessed and a culture-independent detection method for P intermedia/P nigrescens in CF sputum was tested. Results: P intermedia reached cell numbers of >10 ⁵->107 colony-forming units (CFU)/ml sputum. The majority of patients with CF (16/17; 94.1%) produced antibodies against two immunoreactive antigens of P intermedia. Culture supernatant fluids, collected from 10 ⁹ P intermedia cells, were more cytotoxic to respiratory epithelial cells in vitro and inflammatory in mouse lungs than respective fluids from anaerobically grown Ps aeruginosa, while fluids from aerobically grown Ps aeruginosa had the highest cytotoxicity and inflammation. Both pathological effects were largely reduced when culture supernatant fluids from 10⁷ cells of either species were used. P intermedia cells (∼10 ⁶CFU/lung) did not induce mortality in the agar beads lung infection mouse model, while Ps aeruginosa cells caused death in 30% of mice due to rapid multiplication. A P intermedia/P nigrescens-specific PNA probe was significantly more sensitive than culture-dependent diagnostic assays to detect these strict anaerobes. Conclusion: Ps aeruginosa and P intermedia become significantly virulent in vitro and in vivo when cell numbers exceed 108 CFU/lung.

Originalsprog	Engelsk
Tidsskrift	Thorax
Vol/bind	65
Udgave nummer	11
Sider (fra-til)	978-984
Antal sider	7
ISSN	0040-6376
DOI	https://doi.org/10.1136/thx.2010.137745
Status	Udgivet - 1 nov. 2010

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10.1136/thx.2010.137745

Citationsformater

Ulrich, M., Beer, I., Braitmaier, P., Dierkes, M., Kummer, F., Krismer, B., Schumacher, U., Gräpler-Mainka, U., Riethmüller, J., Jensen, P. Ø., Bjarnsholt, T., Høiby, N., Bellon, G., & Döring, G. (2010). Relative contribution of Prevotella intermedia and Pseudomonas aeruginosa to lung pathology in airways of patients with cystic fibrosis. Thorax, 65(11), 978-984. https://doi.org/10.1136/thx.2010.137745

Ulrich, M, Beer, I, Braitmaier, P, Dierkes, M, Kummer, F, Krismer, B, Schumacher, U, Gräpler-Mainka, U, Riethmüller, J, Jensen, PØ , Bjarnsholt, T , Høiby, N, Bellon, G & Döring, G 2010, 'Relative contribution of Prevotella intermedia and Pseudomonas aeruginosa to lung pathology in airways of patients with cystic fibrosis', Thorax, bind 65, nr. 11, s. 978-984. https://doi.org/10.1136/thx.2010.137745

@article{30b6faf3e2654b73a7d1b410bd2fcb8e,

title = "Relative contribution of Prevotella intermedia and Pseudomonas aeruginosa to lung pathology in airways of patients with cystic fibrosis",

abstract = "Background: Patients with cystic fibrosis (CF) with Pseudomonas aeruginosa lung infections produce endobronchial mucus plugs allowing growth of obligate anaerobes including Prevotella spp. Whether obligate anaerobes contribute to the pathophysiology of CF lung disease is unknown. Methods: The virulence of Prevotella intermedia and Ps aeruginosa was investigated in vitro and in mice, antibodies against P intermedia in CF sera were assessed and a culture-independent detection method for P intermedia/P nigrescens in CF sputum was tested. Results: P intermedia reached cell numbers of >10 5->107 colony-forming units (CFU)/ml sputum. The majority of patients with CF (16/17; 94.1%) produced antibodies against two immunoreactive antigens of P intermedia. Culture supernatant fluids, collected from 10 9 P intermedia cells, were more cytotoxic to respiratory epithelial cells in vitro and inflammatory in mouse lungs than respective fluids from anaerobically grown Ps aeruginosa, while fluids from aerobically grown Ps aeruginosa had the highest cytotoxicity and inflammation. Both pathological effects were largely reduced when culture supernatant fluids from 107 cells of either species were used. P intermedia cells (∼10 6CFU/lung) did not induce mortality in the agar beads lung infection mouse model, while Ps aeruginosa cells caused death in 30% of mice due to rapid multiplication. A P intermedia/P nigrescens-specific PNA probe was significantly more sensitive than culture-dependent diagnostic assays to detect these strict anaerobes. Conclusion: Ps aeruginosa and P intermedia become significantly virulent in vitro and in vivo when cell numbers exceed 108 CFU/lung.",

keywords = "Animals, Antibodies, Bacterial, Bacteroidaceae Infections, Bronchoalveolar Lavage Fluid, Cells, Cultured, Colony Count, Microbial, Cystic Fibrosis, Humans, Lung, Mice, Mice, Inbred C57BL, Opportunistic Infections, Prevotella intermedia, Pseudomonas Infections, Pseudomonas aeruginosa, Respiratory Tract Infections, Sputum, Virulence",

author = "Martina Ulrich and Isabelle Beer and Peter Braitmaier and Michaela Dierkes and Florian Kummer and Bernhard Krismer and Ulrike Schumacher and Ute Gr{\"a}pler-Mainka and Joachim Riethm{\"u}ller and Jensen, {Peter {\O}strup} and Thomas Bjarnsholt and Niels H{\o}iby and Gabriel Bellon and Gerd D{\"o}ring",

year = "2010",

month = nov,

day = "1",

doi = "10.1136/thx.2010.137745",

language = "English",

volume = "65",

pages = "978--984",

journal = "Thorax",

issn = "0040-6376",

publisher = "B M J Group",

number = "11",

}

TY - JOUR

T1 - Relative contribution of Prevotella intermedia and Pseudomonas aeruginosa to lung pathology in airways of patients with cystic fibrosis

AU - Ulrich, Martina

AU - Beer, Isabelle

AU - Braitmaier, Peter

AU - Dierkes, Michaela

AU - Kummer, Florian

AU - Krismer, Bernhard

AU - Schumacher, Ulrike

AU - Gräpler-Mainka, Ute

AU - Riethmüller, Joachim

AU - Jensen, Peter Østrup

AU - Bjarnsholt, Thomas

AU - Høiby, Niels

AU - Bellon, Gabriel

AU - Döring, Gerd

PY - 2010/11/1

Y1 - 2010/11/1

N2 - Background: Patients with cystic fibrosis (CF) with Pseudomonas aeruginosa lung infections produce endobronchial mucus plugs allowing growth of obligate anaerobes including Prevotella spp. Whether obligate anaerobes contribute to the pathophysiology of CF lung disease is unknown. Methods: The virulence of Prevotella intermedia and Ps aeruginosa was investigated in vitro and in mice, antibodies against P intermedia in CF sera were assessed and a culture-independent detection method for P intermedia/P nigrescens in CF sputum was tested. Results: P intermedia reached cell numbers of >10 5->107 colony-forming units (CFU)/ml sputum. The majority of patients with CF (16/17; 94.1%) produced antibodies against two immunoreactive antigens of P intermedia. Culture supernatant fluids, collected from 10 9 P intermedia cells, were more cytotoxic to respiratory epithelial cells in vitro and inflammatory in mouse lungs than respective fluids from anaerobically grown Ps aeruginosa, while fluids from aerobically grown Ps aeruginosa had the highest cytotoxicity and inflammation. Both pathological effects were largely reduced when culture supernatant fluids from 107 cells of either species were used. P intermedia cells (∼10 6CFU/lung) did not induce mortality in the agar beads lung infection mouse model, while Ps aeruginosa cells caused death in 30% of mice due to rapid multiplication. A P intermedia/P nigrescens-specific PNA probe was significantly more sensitive than culture-dependent diagnostic assays to detect these strict anaerobes. Conclusion: Ps aeruginosa and P intermedia become significantly virulent in vitro and in vivo when cell numbers exceed 108 CFU/lung.

AB - Background: Patients with cystic fibrosis (CF) with Pseudomonas aeruginosa lung infections produce endobronchial mucus plugs allowing growth of obligate anaerobes including Prevotella spp. Whether obligate anaerobes contribute to the pathophysiology of CF lung disease is unknown. Methods: The virulence of Prevotella intermedia and Ps aeruginosa was investigated in vitro and in mice, antibodies against P intermedia in CF sera were assessed and a culture-independent detection method for P intermedia/P nigrescens in CF sputum was tested. Results: P intermedia reached cell numbers of >10 5->107 colony-forming units (CFU)/ml sputum. The majority of patients with CF (16/17; 94.1%) produced antibodies against two immunoreactive antigens of P intermedia. Culture supernatant fluids, collected from 10 9 P intermedia cells, were more cytotoxic to respiratory epithelial cells in vitro and inflammatory in mouse lungs than respective fluids from anaerobically grown Ps aeruginosa, while fluids from aerobically grown Ps aeruginosa had the highest cytotoxicity and inflammation. Both pathological effects were largely reduced when culture supernatant fluids from 107 cells of either species were used. P intermedia cells (∼10 6CFU/lung) did not induce mortality in the agar beads lung infection mouse model, while Ps aeruginosa cells caused death in 30% of mice due to rapid multiplication. A P intermedia/P nigrescens-specific PNA probe was significantly more sensitive than culture-dependent diagnostic assays to detect these strict anaerobes. Conclusion: Ps aeruginosa and P intermedia become significantly virulent in vitro and in vivo when cell numbers exceed 108 CFU/lung.

KW - Animals

KW - Antibodies, Bacterial

KW - Bacteroidaceae Infections

KW - Bronchoalveolar Lavage Fluid

KW - Cells, Cultured

KW - Colony Count, Microbial

KW - Cystic Fibrosis

KW - Humans

KW - Lung

KW - Mice

KW - Mice, Inbred C57BL

KW - Opportunistic Infections

KW - Prevotella intermedia

KW - Pseudomonas Infections

KW - Pseudomonas aeruginosa

KW - Respiratory Tract Infections

KW - Sputum

KW - Virulence

U2 - 10.1136/thx.2010.137745

DO - 10.1136/thx.2010.137745

M3 - Journal article

C2 - 20880875

SN - 0040-6376

VL - 65

SP - 978

EP - 984

JO - Thorax

JF - Thorax

IS - 11

ER -

Relative contribution of Prevotella intermedia and Pseudomonas aeruginosa to lung pathology in airways of patients with cystic fibrosis

Abstract

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