RESUMEN
The increasing occurrence of antibiotic-resistant Staphylococcus (S.) aureus tremendously limits the antibiotic-based treatment options; therefore, an open discussion of alternative treatment strategies is urgently needed. The use of naturally derived materials might become a more promising concept, not only as directly acting antimicrobials, but also for stimulation of the immune system. Costa Rican plant extracts were screened for their ability to enhance the antimicrobial activity of human blood-derived cells against S. aureus infections. We identified three plant extracts which significantly reduced the growth of S. aureus in the presence of human blood without directly acting as antibacterials: Byrsonima crassifolia acetone bark extract, Mandevilla veraguasensis acetone vine extract and Verbesina oerstediana acetone bark extract (VEOEBA). The effect of VEOEBA was studied in more detail, and revealed that VEOEBA increases the antimicrobial activity of neutrophils by enhancing the formation of neutrophil extracellular traps.
RESUMEN
BACKGROUND: The human leukemia cell line HL-60 is considered an alternative cell culture model to study neutrophil differentiation and migration. The aim of this study was to characterize the suitability of HL-60 cells differentiated to neutrophil-like cells (nHL-60) as substitute for blood-derived human neutrophils to investigate the interaction of neutrophils with Staphylococcus aureus. METHODS: For this purpose, antimicrobial activity, bacterial uptake, production of reactive oxygen species and the release of neutrophil extracellular traps (NETs) by nHL-60 cells were analyzed and compared to primary blood-derived neutrophils using Staphylococcus aureus as important human and animal pathogen. RESULTS: Overall, the antimicrobial activities of nHL-60 cells were distinctly lower compared to blood-derived neutrophils. Furthermore, production of reactive oxygen species as well as NET formation was clearly impaired in nHL-60 cells. CONCLUSION: This study indicates that HL-60 cells are of limited usage as an alternative model to study antimicrobial functions of neutrophils against Staphylococcus aureus.
Asunto(s)
Antiinfecciosos/metabolismo , Neutrófilos/inmunología , Staphylococcus aureus/fisiología , Células Cultivadas , Trampas Extracelulares/efectos de los fármacos , Trampas Extracelulares/metabolismo , Fluorescencia , Células HL-60 , Humanos , Neutrófilos/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Staphylococcus aureus/efectos de los fármacos , Acetato de Tetradecanoilforbol/farmacologíaRESUMEN
Guarea kunthiana is used in folk remedies for the treatment of several diseases including microbial infections. The mechanism behind this phenomenon still needs to be elucidated. Here, we investigated the effect of G. kunthiana bark extract on antimicrobial functions of human and bovine neutrophils as the first line of defense against infections. For this aim, neutrophils were isolated from either human or bovine blood and treated with G. kunthiana bark extract. The antimicrobial activity of the neutrophils against Staphylococcus (S.) aureus and Escherichia (E.) coli was tested in a bacterial survival assay and a fluorescence-based phagocytosis assay. Furthermore, the formation of neutrophil extracellular traps (NETs) was visualized by immunofluorescence microscopy. We show that neutrophils treated with G. kunthiana extract distinctly increased phagocytosis of S. aureus or E. coli. Interestingly, we demonstrate that G. kunthiana bark extract induces the formation of NETs in both cell types. This effect was abolished when treating the cells with diphenyleniodonium chloride (DPI) pointing to a direct implication of the NADPH oxidase-dependent formation of reactive oxygen species in this process. In summary, our data strongly suggest that G. kunthiana bark extract boosts the antimicrobial activities of neutrophils as the first line of defense against invading pathogens.