Bystrzycka Weronika, Moskalik Aneta, Sieczkowska Sandra, Manda-Handzlik Aneta, Demkow Urszula, Ciepiela Olga
Student's Scientific Group at the Department of Laboratory Diagnostics and Clinical Immunology of Developmental Age, Medical University of Warsaw, Poland.
Department of Laboratory Diagnostics and Clinical Immunology of Developmental Age, Medical University of Warsaw, Poland; Postgraduate School of Molecular Medicine, Medical University of Warsaw, Poland.
Cent Eur J Immunol. 2016;41(1):1-5. doi: 10.5114/ceji.2016.58811. Epub 2016 Mar 24.
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.
中性粒细胞胞外陷阱（NETs）是由中性粒细胞释放到细胞外基质中的与抗菌蛋白复合的核DNA细丝，用于结合、固定和杀死不同病原体。NET的形成由不同的生理和非生理刺激物触发。也有人认为抗生素可能是影响NET释放的非生理化合物。本研究的目的是调查克林霉素和阿莫西林对NET释放及中性粒细胞吞噬功能的影响。通过密度离心法从健康供体中分离出的中性粒细胞与阿莫西林或克林霉素孵育两小时，然后用佛波酯12-肉豆蔻酸酯13-乙酸酯（PMA）刺激NET释放。与PMA孵育三小时后，通过荧光法将NETs定量为细胞外DNA的量，并通过免疫荧光显微镜观察。通过流式细胞术测量吞噬细胞的百分比。我们发现，定量测量和荧光显微镜检查证实，阿莫西林可诱导NET形成（细胞外DNA荧光增加，p = 0.03），而克林霉素对NET释放没有影响（p > 0.05）。关于吞噬细胞功能，两种抗生素均增加了细菌摄取（阿莫西林和克林霉素的中位数增加分别为43.3%和61.6%）。我们得出结论，抗生素调节NET释放的能力取决于所用抗生素，且与它们影响吞噬作用的能力无关。
