Methods to study lipid alterations in neutrophils and the subsequent formation of neutrophil extracellular traps

Forskningsoutput: TidskriftsbidragArtikel i vetenskaplig tidskrift

Abstract

Lipid analysis performed by high performance thin layer chromatography (HPTLC) is a relatively simple, cost-effective method of analyzing a broad range of lipids. The function of lipids (e.g., in host-pathogen interactions or host entry) has been reported to play a crucial role in cellular processes. Here, we show a method to determine lipid composition, with a focus on the cholesterol level of primary blood-derived neutrophils, by HPTLC in comparison to high performance liquid chromatography (HPLC). The aim was to investigate the role of lipid/cholesterol alterations in the formation of neutrophil extracellular traps (NETs). NET release is known as a host defense mechanism to prevent pathogens from spreading within the host. Therefore, blood-derived human neutrophils were treated with methyl-β-cyclodextrin (MβCD) to induce lipid alterations in the cells. Using HPTLC and HPLC, we have shown that MβCD treatment of the cells leads to lipid alterations associated with a significant reduction in the cholesterol content of the cell. At the same time, MβCD treatment of the neutrophils led to the formation of NETs, as shown by immunofluorescence microscopy. In summary, here we present a detailed method to study lipid alterations in neutrophils and the formation of NETs.

Detaljer

Författare
  • Graham Brogden
  • Ariane Neumann
  • Diab M. Husein
  • Friederike Reuner
  • Hassan Y. Naim
  • Maren Von Köckritz-Blickwede
Enheter & grupper
Externa organisationer
  • University of Veterinary Medicine Hannover
Forskningsområden

Ämnesklassifikation (UKÄ) – OBLIGATORISK

  • Cell- och molekylärbiologi

Nyckelord

Originalspråkengelska
Artikelnummere54667
TidskriftJournal of Visualized Experiments
Volym2017
Utgåva nummer121
StatusPublished - 2017 mar 29
PublikationskategoriForskning
Peer review utfördJa