Neutrophil extracellular trap formation in supragingival biofilms

被引:58
作者
Hirschfeld, Josefine [1 ]
Dommisch, Henrik [2 ,3 ]
Skora, Philipp [1 ]
Horvath, Gabor [4 ]
Latz, Eicke [4 ]
Hoerauf, Achim [5 ]
Waller, Tobias [1 ]
Kawai, Toshihisa [6 ]
Jepsen, Soren [1 ]
Deschner, James [7 ]
Bekeredjian-Ding, Isabelle [5 ,8 ]
机构
[1] Univ Hosp Bonn, Ctr Dent & Oral Med, Dept Periodontol Operat & Prevent Dent, D-53111 Bonn, Germany
[2] Charite, Dept Periodontol & Synopt Dent, Inst Dent & Oral Med, D-14197 Berlin, Germany
[3] Univ Washington, Dept Oral Hlth Sci, Seattle, WA 98195 USA
[4] Univ Hosp Bonn, Biomed Ctr, Inst Innate Immun, D-53127 Bonn, Germany
[5] Univ Hosp Bonn, Inst Med Microbiol Immunol & Parasitol, D-53127 Bonn, Germany
[6] Forsyth Inst, Dept Immunol & Infect Dis, Cambridge, MA 02142 USA
[7] Univ Hosp Bonn, Dept Expt Dentomaxillofacial Med, Ctr Dent & Oral Med, D-53111 Bonn, Germany
[8] Paul Ehrlich Inst, Div EU Cooperat Microbiol, D-63225 Langen, Germany
关键词
Oral biofilm; Neutrophils; NET; Bacteria; Interleukin-8; STAPHYLOCOCCUS-AUREUS BIOFILMS; GINGIVAL CREVICULAR FLUID; HOST-DEFENSE; CATHEPSIN-G; ANTIMICROBIAL PEPTIDES; INNATE IMMUNITY; CHRONIC PERIODONTITIS; CATHELICIDIN LL-37; SALIVARY LEVELS; DISEASE;
D O I
10.1016/j.ijmm.2015.04.002
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Background: Oral biofilms are the causative agents of the highly prevalent oral diseases periodontitis and caries. Additionally, the host immune response is thought to play a critical role in disease onset. Neutrophils are known to be a key host response factor to bacterial challenge on host surfaces. Release of neutrophil extracellular traps (NETs) as a novel antimicrobial defense strategy has gained increasing attention in the past years. Here, we investigated the influx of neutrophils into the dental plaque and the ability of oral bacteria to trigger intra-biofilm release of NETs and intracellular proteins. Methods: Supragingival biofilms and whole saliva were sampled from systemically healthy subjects participating in an experimental gingivitis study. Biofilms were analysed by immunofluorescence followed by confocal and fluorescence microscopy. Moreover, concentrations of cytokines and immune-associated proteins in biofilm suspensions and saliva were assessed by ELISA. Neutrophils obtained from blood were stimulated with twelve bacterial species isolated from cultured biofilms or with lipopolysaccharide to monitor NET formation. Results: Neutrophils, NETs, neutrophil-associated proteins (myeloperoxidase, elastase-2, cathepsin G, cathelicidin LL-37), interleukin-8, interleukin-1 beta and tumor necrosis factor were detected within plaque samples and saliva. All tested bacterial species as well as the polymicrobial samples isolated from the plaque of each donor induced release of NETs and interleukin-8. The degree of NET formation varied among different subjects and did not correlate with plaque scores or clinical signs of local inflammation. Conclusions: Our findings indicate that neutrophils are attracted towards dental biofilms, in which they become incorporated and where they are stimulated by microbes to release NETs and immunostimulatory proteins. Thus, neutrophils and NETs may be involved in host biofilm control, although their specific role needs to be further elucidated. Moreover, inter-patient variability suggests NET formation as a potential factor influencing the individual course of disease. (C) 2015 Elsevier GmbH. All rights reserved.
引用
收藏
页码:453 / 463
页数:11
相关论文
共 83 条
[1]   Elevated neutrophil membrane expression of proteinase 3 is dependent upon CD177 expression [J].
Abdgawad, M. ;
Gunnarsson, L. ;
Bengtsson, A. A. ;
Geborek, P. ;
Nilsson, L. ;
Segelmark, M. ;
Hellmark, T. .
CLINICAL AND EXPERIMENTAL IMMUNOLOGY, 2010, 161 (01) :89-97
[2]   Infections in patients with inherited defects in phagocytic function [J].
Andrews, T ;
Sullivan, KE .
CLINICAL MICROBIOLOGY REVIEWS, 2003, 16 (04) :597-+
[3]   The Oral Microbiota: Living with a Permanent Guest [J].
Avila, Maria ;
Ojcius, David M. ;
Yilmaz, Oezlem .
DNA AND CELL BIOLOGY, 2009, 28 (08) :405-411
[4]   INTERLEUKIN-8, A CHEMOTACTIC AND INFLAMMATORY CYTOKINE [J].
BAGGIOLINI, M ;
CLARKLEWIS, I .
FEBS LETTERS, 1992, 307 (01) :97-101
[5]   The Balancing Act of Neutrophils [J].
Bardoel, Bart W. ;
Kenny, Elaine F. ;
Sollberger, Gabriel ;
Zychlinsky, Arturo .
CELL HOST & MICROBE, 2014, 15 (05) :526-536
[6]  
BARKER JNWN, 1991, AM J PATHOL, V139, P869
[7]   Periodontitis: a host-mediated disruption of microbial homeostasis. Unlearning learned concepts [J].
Bartold, P. Mark ;
Van Dyke, Thomas E. .
PERIODONTOLOGY 2000, 2013, 62 :203-217
[8]   Impact of Extracellular Nuclease Production on the Biofilm Phenotype of Staphylococcus aureus under In Vitro and In Vivo Conditions [J].
Beenken, Karen E. ;
Spencer, Horace ;
Griffin, Linda M. ;
Smeltzer, Mark S. .
INFECTION AND IMMUNITY, 2012, 80 (05) :1634-1638
[9]  
Boronat-Catala Montserrat, 2014, J Clin Exp Dent, V6, pe175, DOI 10.4317/jced.51403
[10]   Neutrophil extracellular traps kill bacteria [J].
Brinkmann, V ;
Reichard, U ;
Goosmann, C ;
Fauler, B ;
Uhlemann, Y ;
Weiss, DS ;
Weinrauch, Y ;
Zychlinsky, A .
SCIENCE, 2004, 303 (5663) :1532-1535