This video demonstrates a procedure for the adoptive transfer of high salt-treated antigen-presenting dendritic cells (DCs) into mice. Murine CD11c+ dendritic cells (DCs) are exposed to a medium containing high amounts of sodium, forming isolevuglandin (IsoLG)-protein adducts in the cells. These adducts serve as neoantigens and are presented on the surface of DCs. The high salt-treated cells are injected into mice through adoptive cell transfer.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Isolation of Spleens from Mice Prepare 1640 RPMI (Roswell Park Memorial institute): 10% fetal bovine serum (FBS), 0.10 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), 1 mM sodium pyruvate, 50 µM β-mercaptoethanol and 1% penicillin/streptomycin. Euthanize 10–12 week-old C57bl/6 ma…
A Method for the High Salt Treatment of Dendritic Cells and their Adoptive Transfer into Mice. J. Vis. Exp. (Pending Publication), e22277, doi: (2024).