In Vitro Analysis of Nanoparticle Effects on the Zymosan Uptake by Phagocytic Cells

Methods Mol Biol. 2018;1682:125-133. doi: 10.1007/978-1-4939-7352-1_11.

Abstract

This chapter provides a protocol for analysis of nanoparticle effects on the function of phagocytic cells. The protocol relies on luminol chemiluminescence to detect zymosan uptake. Zymosan is an yeast particle which is typically eliminated by phagocytic cells via the complement receptor pathway. The luminol, co-internalized with zymosan, is processed inside the phagosome to generate a chemiluminescent signal. If a test nanoparticle affects the phagocytic function of the cell, the amount of phagocytosed zymosan and, proportionally, the level of generated chemiluminescent signal change. Comparing the zymosan uptake of untreated cells with that of cells exposed to a nanoparticle provides information about the nanoparticle's effects on the normal phagocytic function. This method has been described previously and is presented herein with several changes. The revised method includes details about nanoparticle concentration selection, updated experimental procedure, and examples of the method performance.

Keywords: Immunosuppression; Nanoparticle; Phagocytosis; Zymosan A.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • HL-60 Cells
  • Humans
  • Luminescent Measurements / methods*
  • Luminol / analysis
  • Phagocytes / cytology*
  • Phagocytes / immunology
  • Phagocytosis*
  • Zymosan / analysis*
  • Zymosan / immunology

Substances

  • Luminol
  • Zymosan