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2016 ; 8
(ä): 33070
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Effects of Aggregatibacter actinomycetemcomitans leukotoxin on neutrophil
migration and extracellular trap formation
#MMPMID27834173
Hirschfeld J
; Roberts HM
; Chapple IL
; Par?ina M
; Jepsen S
; Johansson A
; Claesson R
J Oral Microbiol
2016[]; 8
(ä): 33070
PMID27834173
show ga
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.