Isolation and Adoptive Transfer of High Salt Treated Antigen-presenting Dendritic Cells

J Vis Exp. 2019 Mar 5:(145). doi: 10.3791/59124.

Abstract

Excess dietary salt intake contributes to inflammation and plays a vital role in the development of hypertension. We previously found that antigen-presenting dendritic cells (DCs) can sense elevated extracellular sodium leading to the activation of the NADPH oxidase and formation of isolevuglandin (IsoLG)-protein adducts. These IsoLG-protein adducts react with self-proteins and promote an autoimmune-like state and hypertension. We have developed and optimized state-of-the-art methods to study DC function in hypertension. Here, we provide a detailed protocol for isolation, in vitro treatment with elevated sodium, and adoptive transfer of murine splenic CD11c+ cells into recipient mice to study their role in hypertension.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Adoptive Transfer*
  • Animals
  • Antigen-Presenting Cells / cytology*
  • Antigen-Presenting Cells / drug effects
  • Blood Pressure / drug effects
  • CD11c Antigen / metabolism
  • Dendritic Cells / cytology*
  • Dendritic Cells / drug effects
  • Hypertension
  • Lipids / pharmacology
  • Magnetic Phenomena
  • Male
  • Mice, Inbred C57BL
  • Sodium Chloride / pharmacology*
  • Spleen / cytology
  • Systole / drug effects

Substances

  • CD11c Antigen
  • Lipids
  • isolevuglandin
  • Sodium Chloride