Limits...
The effect of clindamycin and amoxicillin on neutrophil extracellular trap (NET) release.

Bystrzycka W, Moskalik A, Sieczkowska S, Manda-Handzlik A, Demkow U, Ciepiela O - Cent Eur J Immunol (2016)

Bottom Line: We showed that amoxicillin induces NET formation (increase of extracellular DNA fluorescence, p = 0.03), while clindamycin had no influence on NET release (p > 0.05), as confirmed by quantitative measurement and fluorescent microscopy.Regarding phagocyte function, both antibiotics increased bacterial uptake (43.3% and 61.6% median increase for amoxicillin and clindamycin, respectively).We concluded that the ability of antibiotics to modulate NET release depends on the antibiotic used and is not associated with their ability to influence phagocytosis.

View Article: PubMed Central - PubMed

Affiliation: Student's Scientific Group at the Department of Laboratory Diagnostics and Clinical Immunology of Developmental Age, Medical University of Warsaw, Poland.

ABSTRACT
Neutrophil extracellular traps (NETs) are threads of nuclear DNA complexed with antimicrobial proteins released by neutrophils to extracellular matrix to bind, immobilise, and kill different pathogens. NET formation is triggered by different physiological and non-physiological stimulants. It is also suggested that antibiotics could be non-physiological compounds that influence NET release. The aim of the study was to investigate the effect of clindamycin and amoxicillin on NET release and the phagocyte function of neutrophils. Neutrophils isolated from healthy donors by density centrifugation method were incubated with amoxicillin or clindamycin for two hours, and then NET release was stimulated with phorbol 12-myristate 13-acetate (PMA). After three hours of incubation with PMA NETs were quantified as amount of extracellular DNA by fluorometry and visualised by immunofluorescent microscopy. The percent of phagocyting cells was measured by flow cytometry. We showed that amoxicillin induces NET formation (increase of extracellular DNA fluorescence, p = 0.03), while clindamycin had no influence on NET release (p > 0.05), as confirmed by quantitative measurement and fluorescent microscopy. Regarding phagocyte function, both antibiotics increased bacterial uptake (43.3% and 61.6% median increase for amoxicillin and clindamycin, respectively). We concluded that the ability of antibiotics to modulate NET release depends on the antibiotic used and is not associated with their ability to influence phagocytosis.

No MeSH data available.


Related in: MedlinePlus

The amount of extracellular DNA released from antibiotic- pretreated neutrophils without or after stimulation with 100 nM PMA. Amoxicillin pretreated cells showed increased release of NETs, while clindamycin did not affect NET release. Samples treated only with antibiotics served as negative control for NET formation
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4829816&req=5

Figure 0002: The amount of extracellular DNA released from antibiotic- pretreated neutrophils without or after stimulation with 100 nM PMA. Amoxicillin pretreated cells showed increased release of NETs, while clindamycin did not affect NET release. Samples treated only with antibiotics served as negative control for NET formation

Mentions: The quantity of released DNA was analysed using a fluorescence reader. The results show that clindamycin does not affect PMA-induced NET formation (78,628 ±17,290 rfu for PMA-stimulated sample vs. 78,628 ±10,249 rfu for amoxicillin-pretreated, PMA-stimulated sample p = 0.2), while amoxicillin increases NETS release (59,972 ±9755 rfu for PMA-stimulated sample and 74,306 ±9519 rfu for amoxicillin-pretreated, PMA-stimulated sample, p = 0.03) (Fig. 2). Neutrophil extracellular traps visualisation using immunofluorescent microscopy confirmed these observations (Fig. 3).


The effect of clindamycin and amoxicillin on neutrophil extracellular trap (NET) release.

Bystrzycka W, Moskalik A, Sieczkowska S, Manda-Handzlik A, Demkow U, Ciepiela O - Cent Eur J Immunol (2016)

The amount of extracellular DNA released from antibiotic- pretreated neutrophils without or after stimulation with 100 nM PMA. Amoxicillin pretreated cells showed increased release of NETs, while clindamycin did not affect NET release. Samples treated only with antibiotics served as negative control for NET formation
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4829816&req=5

Figure 0002: The amount of extracellular DNA released from antibiotic- pretreated neutrophils without or after stimulation with 100 nM PMA. Amoxicillin pretreated cells showed increased release of NETs, while clindamycin did not affect NET release. Samples treated only with antibiotics served as negative control for NET formation
Mentions: The quantity of released DNA was analysed using a fluorescence reader. The results show that clindamycin does not affect PMA-induced NET formation (78,628 ±17,290 rfu for PMA-stimulated sample vs. 78,628 ±10,249 rfu for amoxicillin-pretreated, PMA-stimulated sample p = 0.2), while amoxicillin increases NETS release (59,972 ±9755 rfu for PMA-stimulated sample and 74,306 ±9519 rfu for amoxicillin-pretreated, PMA-stimulated sample, p = 0.03) (Fig. 2). Neutrophil extracellular traps visualisation using immunofluorescent microscopy confirmed these observations (Fig. 3).

Bottom Line: We showed that amoxicillin induces NET formation (increase of extracellular DNA fluorescence, p = 0.03), while clindamycin had no influence on NET release (p > 0.05), as confirmed by quantitative measurement and fluorescent microscopy.Regarding phagocyte function, both antibiotics increased bacterial uptake (43.3% and 61.6% median increase for amoxicillin and clindamycin, respectively).We concluded that the ability of antibiotics to modulate NET release depends on the antibiotic used and is not associated with their ability to influence phagocytosis.

View Article: PubMed Central - PubMed

Affiliation: Student's Scientific Group at the Department of Laboratory Diagnostics and Clinical Immunology of Developmental Age, Medical University of Warsaw, Poland.

ABSTRACT
Neutrophil extracellular traps (NETs) are threads of nuclear DNA complexed with antimicrobial proteins released by neutrophils to extracellular matrix to bind, immobilise, and kill different pathogens. NET formation is triggered by different physiological and non-physiological stimulants. It is also suggested that antibiotics could be non-physiological compounds that influence NET release. The aim of the study was to investigate the effect of clindamycin and amoxicillin on NET release and the phagocyte function of neutrophils. Neutrophils isolated from healthy donors by density centrifugation method were incubated with amoxicillin or clindamycin for two hours, and then NET release was stimulated with phorbol 12-myristate 13-acetate (PMA). After three hours of incubation with PMA NETs were quantified as amount of extracellular DNA by fluorometry and visualised by immunofluorescent microscopy. The percent of phagocyting cells was measured by flow cytometry. We showed that amoxicillin induces NET formation (increase of extracellular DNA fluorescence, p = 0.03), while clindamycin had no influence on NET release (p > 0.05), as confirmed by quantitative measurement and fluorescent microscopy. Regarding phagocyte function, both antibiotics increased bacterial uptake (43.3% and 61.6% median increase for amoxicillin and clindamycin, respectively). We concluded that the ability of antibiotics to modulate NET release depends on the antibiotic used and is not associated with their ability to influence phagocytosis.

No MeSH data available.


Related in: MedlinePlus