Isolation protocol of mouse monocyte-derived dendritic cells and their subsequent in vitro activation with tumor immune complexes

Nadine Santana-Magal, Diana Rasoulouniriana, Corey Saperia, Amit Gutwillig, Peleg Rider, Edgar G. Engleman, Yaron Carmi

Research output: Contribution to journalArticlepeer-review

Abstract

Dendritic cells (DC) are heterogeneous cell populations that differ in their cell membrane markers, migration patterns and distribution, and in their antigen presentation and T cell activation capacities. Since most vaccinations of experimental tumor models require millions of DC, they are widely isolated from the bone marrow or spleen. However, these DC significantly differ from blood and tumor DC in their responses to immune complexes (IC), and presumably to other Syk-coupled lectin receptors. Importantly, given the sensitivity of DC to danger-associated molecules, the presence of endotoxins or antibodies that crosslink activation receptors in one of the isolating steps could result in the priming of DC and thus affect the parameters, or at least the dosage, required to activate them. Therefore, here we describe a detailed protocol for isolating MoDC from blood and tumors while avoiding their premature activation. In addition, a protocol is provided for MoDC activation with tumor IC, and their subsequent analyses.

Original languageEnglish
Article numbere57188
JournalJournal of Visualized Experiments
Volume2018
Issue number135
DOIs
StatePublished - 31 May 2018

Keywords

  • Cancer research
  • Dendritic cells
  • Fcγ receptors
  • Immune complexes
  • Issue 135
  • MAP kinases
  • Monocyte-derived DC
  • Tumor-associated DC
  • Tyrosine phosphatase

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