Detecting Migration and Infiltration of Neutrophils in Mice

J Vis Exp. 2020 Feb 6:(156). doi: 10.3791/60543.

Abstract

Neutrophils are a major member of the innate immune system and play pivotal roles in host defense against pathogens and pathologic inflammatory reactions. Neutrophils can be recruited to inflammation sites via the guidance of cytokines and chemokines. Overwhelming infiltration of neutrophils can lead to indiscriminate tissue damage, such as in rheumatoid arthritis (RA). Neutrophils isolated from peritoneal exudate respond to a defined chemoattractant, N-formyl-Met-Leu-Phe (fMLP), in vitro in Transwell or Zigmond chamber assays. The air pouch experiment can be used to evaluate the chemotaxis of neutrophils towards lipopolysaccharide (LPS) in vivo. The adjuvant-induced arthritis (AA) mouse model is frequently used in RA research, and immunohistochemical staining of joint sections with anti-myeloperoxidase (MPO) or anti-neutrophil elastase (NE) antibodies is a well-established method to measure neutrophil infiltration. These methods can be used to discover promising therapies targeting neutrophil migration.

Publication types

  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Animals
  • Arthritis, Experimental / immunology
  • Arthritis, Experimental / pathology
  • Cell Movement* / drug effects
  • Cell Separation
  • Disease Models, Animal
  • Joints / pathology
  • Mice, Inbred C57BL
  • N-Formylmethionine Leucyl-Phenylalanine / analogs & derivatives
  • N-Formylmethionine Leucyl-Phenylalanine / pharmacology
  • Neutrophil Infiltration / drug effects
  • Neutrophil Infiltration / immunology
  • Neutrophils / cytology*
  • Neutrophils / drug effects
  • Neutrophils / immunology

Substances

  • N-Formylmethionine Leucyl-Phenylalanine
  • methionyl-leucyl-phenylalanine