Eimeria ninakohlyakimovae induces NADPH oxidase-dependent monocyte extracellular trap formation and upregulates IL-12 and TNF-α, IL-6 and CCL2 gene transcription

Vet Parasitol. 2016 Aug 30:227:143-50. doi: 10.1016/j.vetpar.2016.07.028. Epub 2016 Jul 25.

Abstract

Extracellular trap (ET) formation has been demonstrated as novel effector mechanism against diverse pathogens in polymorphonuclear neutrophils (PMN), eosinophils, mast cells, macrophages and recently also in monocytes. In the current study, we show that E. ninakohlyakimovae triggers the deliverance of monocyte-derived ETs in vitro. Fluorescence illustrations as well as scanning electron microscopy (SEM) analyses showed that monocyte-derived ET formation was rapidly induced upon exposure to viable sporozoites, sporocysts and oocysts of E. ninakohlyakimovae. Classical features of monocyte-released ETs were confirmed by the co-localization of extracellular DNA adorned with myeloperoxidase (MPO) and histones (H3) in parasite-entrapping structures. The treatment of caprine monocyte ET structures with NADPH oxidase inhibitor diphenylene iodondium (DPI) significantly reduced ETosis confirming the essential role of reactive oxygen species (ROS) in monocyte mediated ETs formation. Additionally, co-culture of monocytes with viable sporozoites and soluble oocyst antigen (SOA) induced distinct levels of cytokine and chemokine gene transcription. Thus, the transcription of genes encoding for IL-12 and TNF-α was significantly upregulated after sporozoite encounter. In contrast IL-6 and CCL2 gene transcripts were rather weakly induced by parasites. Conversely, SOA only induced the up-regulation of IL-6 and CCL2 gene transcription, and failed to enhance transcripts of IL-12 and TNF-α in vitro. We here report on monocyte-triggered ETs as novel effector mechanism against E. ninakohlyakimovae. Our results strongly suggest that monocyte-mediated innate immune reactions might play an important role in early host immune reactions against E. ninakohlyakimovae in goats.

Keywords: Chemokines; Cytokines; Eimeria ninakohlyakimovae; Extracellular traps; Innate immune reactions; Monocyte.

MeSH terms

  • Animals
  • Cells, Cultured
  • Chemokine CCL2 / genetics
  • Chemokine CCL2 / metabolism
  • Cytokines / genetics
  • Cytokines / metabolism*
  • Eimeria / classification*
  • Gene Expression Regulation / immunology*
  • Goat Diseases / parasitology
  • Goats
  • Interleukin-12 / genetics
  • Interleukin-12 / metabolism
  • Interleukin-6 / genetics
  • Interleukin-6 / metabolism
  • Monocytes / physiology*
  • Monocytes / ultrastructure
  • NADPH Oxidases / genetics
  • NADPH Oxidases / metabolism*
  • Tumor Necrosis Factor-alpha / genetics
  • Tumor Necrosis Factor-alpha / metabolism
  • Up-Regulation

Substances

  • Chemokine CCL2
  • Cytokines
  • Interleukin-6
  • Tumor Necrosis Factor-alpha
  • Interleukin-12
  • NADPH Oxidases