Homeostasis inside Single Activated Phagolysosomes: Quantitative and Selective Measurements of Submillisecond Dynamics of Reactive Oxygen and Nitrogen Species Production with a Nanoelectrochemical Sensor

J Am Chem Soc. 2022 Jun 8;144(22):9723-9733. doi: 10.1021/jacs.2c01857. Epub 2022 May 26.

Abstract

Reactive oxygen and nitrogen species (ROS/RNS) are generated by macrophages inside their phagolysosomes. This production is essential for phagocytosis of damaged cells and pathogens, i.e., protecting the organism and maintaining immune homeostasis. The ability to quantitatively and individually monitor the four primary ROS/RNS (ONOO-, H2O2, NO, and NO2-) with submillisecond resolution is clearly warranted to elucidate the still unclear mechanisms of their rapid generation and to track their concentration variations over time inside phagolysosomes, in particular, to document the origin of ROS/RNS homeostasis during phagocytosis. A novel nanowire electrode has been specifically developed for this purpose. It consisted of wrapping a SiC nanowire with a mat of 3 nm platinum nanoparticles whose high electrocatalytic performances allow the characterization and individual measurements of each of the four primary ROS/RNS. This allowed, for the first time, a quantitative, selective, and statistically robust determination of the individual amounts of ROS/RNS present in single dormant phagolysosomes. Additionally, the submillisecond resolution of the nanosensor allowed confirmation and measurement of the rapid ability of phagolysosomes to differentially mobilize their enzyme pools of NADPH oxidases and inducible nitric oxide synthases to finely regulate their homeostasis. This reveals an essential key to immune responses and immunotherapies and rationalizes its biomolecular origin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Homeostasis
  • Hydrogen Peroxide
  • Metal Nanoparticles*
  • Nitrogen
  • Oxygen*
  • Phagosomes
  • Platinum
  • Reactive Nitrogen Species / chemistry
  • Reactive Oxygen Species / chemistry

Substances

  • Reactive Nitrogen Species
  • Reactive Oxygen Species
  • Platinum
  • Hydrogen Peroxide
  • Nitrogen
  • Oxygen