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Article

Methylprednisolone Induces Extracellular Trap Formation and Enhances Bactericidal Effect of Canine Neutrophils

1
Department of Small Animal Medicine and Surgery, University of Veterinary Medicine Hannover, Foundation, 30559 Hannover, Germany
2
Department of Biochemistry, University of Veterinary Medicine Hannover, Foundation, 30559 Hannover, Germany
3
Research Center for Emerging Infections and Zoonoses, University of Veterinary Medicine Hannover, Foundation, 30559 Hannover, Germany
4
Centre for Infection Medicine, Institute of Microbiology and Epizootics, Freie Universität Berlin, 14163 Berlin, Germany
*
Authors to whom correspondence should be addressed.
Academic Editor: Milladur Rahman
Int. J. Mol. Sci. 2021, 22(14), 7734; https://doi.org/10.3390/ijms22147734
Received: 28 April 2021 / Revised: 11 July 2021 / Accepted: 13 July 2021 / Published: 20 July 2021
(This article belongs to the Special Issue Molecular Research on Neutrophil Extracellular Trap (NET))
Methylprednisolone is a glucocorticoid and can negatively influence immune defense mechanisms. During bacterial infections in the dog, neutrophils infiltrate infected tissue and mediate antimicrobial effects with different mechanisms such as phagocytosis and neutrophil extracellular trap (NET) formation. Here, we investigated the influence of methylprednisolone on canine NET formation and neutrophil killing efficiency of Gram positive and Gram negative bacteria. Therefore, canine blood derived neutrophils were treated with different concentrations of methylprednisolone over time. The survival factor of Staphylococcus pseudintermedius, Streptococcus canis or Escherichia coli was determined in presence of stimulated neutrophils. Additionally, free DNA and nucleosomes as NET marker were analyzed in supernatants and neutrophils were assessed for NET formation by immunofluorescence microscopy. Methylprednisolone concentrations of 62.5 and 625 µg/mL enhanced the neutrophil killing of Gram positive bacteria, whereas no significant influence was detected for the Gram negative Escherichia coli. Interestingly, higher amounts of free DNA were detected under methylprednisolone stimulation in a concentration dependency and in the presence of Streptococcus canis and Escherichia coli. The nucleosome release by neutrophils is induced by bacterial infection and differs depending on the concentration of methylprednisolone. Furthermore, immunofluorescence microscopy analysis identified methylprednisolone at a concentration of 62.5 µg/mL as a NET inducer. In summary, methylprednisolone enhances NET-formation and time-dependent and concentration-dependent the bactericidal effect of canine neutrophils on Gram positive bacteria. View Full-Text
Keywords: NETs; methylprednisolone; free DNA; dog; Gram negative bacteria; Gram positive bacteria NETs; methylprednisolone; free DNA; dog; Gram negative bacteria; Gram positive bacteria
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MDPI and ACS Style

Steffensen, N.; Imker, R.; Lassnig, S.; Fulde, M.; Rieder, J.C.; de Buhr, N. Methylprednisolone Induces Extracellular Trap Formation and Enhances Bactericidal Effect of Canine Neutrophils. Int. J. Mol. Sci. 2021, 22, 7734. https://doi.org/10.3390/ijms22147734

AMA Style

Steffensen N, Imker R, Lassnig S, Fulde M, Rieder JC, de Buhr N. Methylprednisolone Induces Extracellular Trap Formation and Enhances Bactericidal Effect of Canine Neutrophils. International Journal of Molecular Sciences. 2021; 22(14):7734. https://doi.org/10.3390/ijms22147734

Chicago/Turabian Style

Steffensen, Nicole, Rabea Imker, Simon Lassnig, Marcus Fulde, Johanna C. Rieder, and Nicole de Buhr. 2021. "Methylprednisolone Induces Extracellular Trap Formation and Enhances Bactericidal Effect of Canine Neutrophils" International Journal of Molecular Sciences 22, no. 14: 7734. https://doi.org/10.3390/ijms22147734

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