Effects of Aggregatibacter actinomycetemcomitans leukotoxin on neutrophil migration and extracellular trap formation
Background: Aggressive periodontitis is associated with the presence of Aggregatibacter actinomycetemcomitans, a leukotoxin (Ltx)-producing periodontal pathogen. Ltx has the ability to lyse white blood cells including neutrophils. Objectives: This study was aimed at investigating the interactions be...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2016-11-01
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| Series: | Journal of Oral Microbiology |
| Subjects: | |
| Online Access: | http://www.journaloforalmicrobiology.net/index.php/jom/article/view/33070/49731 |
| _version_ | 1818846712713707520 |
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| author | Josefine Hirschfeld Helen M. Roberts Iain L. C. Chapple Marijo Parčina Søren Jepsen Anders Johansson Rolf Claesson |
| author_facet | Josefine Hirschfeld Helen M. Roberts Iain L. C. Chapple Marijo Parčina Søren Jepsen Anders Johansson Rolf Claesson |
| author_sort | Josefine Hirschfeld |
| collection | DOAJ |
| description | Background: Aggressive periodontitis is associated with the presence of Aggregatibacter actinomycetemcomitans, a leukotoxin (Ltx)-producing periodontal pathogen. Ltx has the ability to lyse white blood cells including neutrophils. Objectives: This study was aimed at investigating the interactions between neutrophils and Ltx with regard to the chemotactic properties of Ltx and the release of neutrophil extracellular traps (NETs). Methods: Neutrophils from healthy blood donors were isolated and incubated for 30 min and 3 h with increasing concentrations of Ltx (1, 10, and 100 ng/mL) as well as with A. actinomycetemcomitans strains (NCTC 9710 and HK 1651) producing different levels of Ltx. Formation of NETs and cell lysis were assessed by microscopy, fluorescence-based assays, and measurement of released lactate dehydrogenase. Neutrophil migration in response to different Ltx gradients was monitored by real-time video microscopy, and image analysis was performed using ImageJ software. Results: Although Ltx (10 and 100 ng/mL) and the leukotoxic A. actinomycetemcomitans strain HK 1651 lysed some neutrophils, other cells were still capable of performing NETosis in a concentration-dependent manner. Low doses of Ltx and the weakly leukotoxic strain NCTC 9710 did not lead to neutrophil lysis, but did induce some NETosis. Furthermore, all three concentrations of Ltx enhanced random neutrophil movement; however, low directional accuracy was observed compared with the positive control (fMLP). Conclusions: The results indicate that Ltx acts both as a neutrophil activator and also causes cell death. In addition, Ltx directly induces NETosis in neutrophils prior to cell lysis. In future studies, the underlying pathways involved in Ltx-meditated neutrophil activation and NETosis need to be investigated further. |
| first_indexed | 2024-12-19T05:49:55Z |
| format | Article |
| id | doaj.art-a04f4f4e9f88413994a09410c853e3e0 |
| institution | Directory Open Access Journal |
| issn | 2000-2297 |
| language | English |
| last_indexed | 2024-12-19T05:49:55Z |
| publishDate | 2016-11-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Journal of Oral Microbiology |
| spelling | doaj.art-a04f4f4e9f88413994a09410c853e3e02022-12-21T20:33:40ZengTaylor & Francis GroupJournal of Oral Microbiology2000-22972016-11-01801810.3402/jom.v8.3307033070Effects of Aggregatibacter actinomycetemcomitans leukotoxin on neutrophil migration and extracellular trap formationJosefine Hirschfeld0Helen M. Roberts1Iain L. C. Chapple2Marijo Parčina3Søren Jepsen4Anders Johansson5Rolf Claesson6 Periodontal Research Group, Birmingham Dental School & Hospital, Birmingham, United Kingdom Periodontal Research Group, Birmingham Dental School & Hospital, Birmingham, United Kingdom Periodontal Research Group, Birmingham Dental School & Hospital, Birmingham, United Kingdom Institute of Medical Microbiology, Immunology and Parasitology, University Hospital Bonn, Bonn, Germany Department of Periodontology, Operative and Preventive Dentistry, University Hospital Bonn, Bonn, Germany Section Molecular Periodontology, Department of Odontology, Umeå University, Umeå, Sweden Section Oral Microbiology, Department of Odontology, Umeå University, Umeå, SwedenBackground: Aggressive periodontitis is associated with the presence of Aggregatibacter actinomycetemcomitans, a leukotoxin (Ltx)-producing periodontal pathogen. Ltx has the ability to lyse white blood cells including neutrophils. Objectives: This study was aimed at investigating the interactions between neutrophils and Ltx with regard to the chemotactic properties of Ltx and the release of neutrophil extracellular traps (NETs). Methods: Neutrophils from healthy blood donors were isolated and incubated for 30 min and 3 h with increasing concentrations of Ltx (1, 10, and 100 ng/mL) as well as with A. actinomycetemcomitans strains (NCTC 9710 and HK 1651) producing different levels of Ltx. Formation of NETs and cell lysis were assessed by microscopy, fluorescence-based assays, and measurement of released lactate dehydrogenase. Neutrophil migration in response to different Ltx gradients was monitored by real-time video microscopy, and image analysis was performed using ImageJ software. Results: Although Ltx (10 and 100 ng/mL) and the leukotoxic A. actinomycetemcomitans strain HK 1651 lysed some neutrophils, other cells were still capable of performing NETosis in a concentration-dependent manner. Low doses of Ltx and the weakly leukotoxic strain NCTC 9710 did not lead to neutrophil lysis, but did induce some NETosis. Furthermore, all three concentrations of Ltx enhanced random neutrophil movement; however, low directional accuracy was observed compared with the positive control (fMLP). Conclusions: The results indicate that Ltx acts both as a neutrophil activator and also causes cell death. In addition, Ltx directly induces NETosis in neutrophils prior to cell lysis. In future studies, the underlying pathways involved in Ltx-meditated neutrophil activation and NETosis need to be investigated further.http://www.journaloforalmicrobiology.net/index.php/jom/article/view/33070/49731leukotoxinAggregatibacter actinomycetemcomitansneutrophilsNETsmigrationchemotaxisaggressive periodontitis |
| spellingShingle | Josefine Hirschfeld Helen M. Roberts Iain L. C. Chapple Marijo Parčina Søren Jepsen Anders Johansson Rolf Claesson Effects of Aggregatibacter actinomycetemcomitans leukotoxin on neutrophil migration and extracellular trap formation Journal of Oral Microbiology leukotoxin Aggregatibacter actinomycetemcomitans neutrophils NETs migration chemotaxis aggressive periodontitis |
| title | Effects of Aggregatibacter actinomycetemcomitans leukotoxin on neutrophil migration and extracellular trap formation |
| title_full | Effects of Aggregatibacter actinomycetemcomitans leukotoxin on neutrophil migration and extracellular trap formation |
| title_fullStr | Effects of Aggregatibacter actinomycetemcomitans leukotoxin on neutrophil migration and extracellular trap formation |
| title_full_unstemmed | Effects of Aggregatibacter actinomycetemcomitans leukotoxin on neutrophil migration and extracellular trap formation |
| title_short | Effects of Aggregatibacter actinomycetemcomitans leukotoxin on neutrophil migration and extracellular trap formation |
| title_sort | effects of aggregatibacter actinomycetemcomitans leukotoxin on neutrophil migration and extracellular trap formation |
| topic | leukotoxin Aggregatibacter actinomycetemcomitans neutrophils NETs migration chemotaxis aggressive periodontitis |
| url | http://www.journaloforalmicrobiology.net/index.php/jom/article/view/33070/49731 |
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