Skip to Content
Merck
  • Cellular Differentiation of Human Monocytes Is Regulated by Time-Dependent Interleukin-4 Signaling and the Transcriptional Regulator NCOR2.

Cellular Differentiation of Human Monocytes Is Regulated by Time-Dependent Interleukin-4 Signaling and the Transcriptional Regulator NCOR2.

Immunity (2017-12-21)
Jil Sander, Susanne V Schmidt, Branko Cirovic, Naomi McGovern, Olympia Papantonopoulou, Anna-Lena Hardt, Anna C Aschenbrenner, Christoph Kreer, Thomas Quast, Alexander M Xu, Lisa M Schmidleithner, Heidi Theis, Lan Do Thi Huong, Hermi Rizal Bin Sumatoh, Mario A R Lauterbach, Jonas Schulte-Schrepping, Patrick Günther, Jia Xue, Kevin Baßler, Thomas Ulas, Kathrin Klee, Natalie Katzmarski, Stefanie Herresthal, Wolfgang Krebs, Bianca Martin, Eicke Latz, Kristian Händler, Michael Kraut, Waldemar Kolanus, Marc Beyer, Christine S Falk, Bettina Wiegmann, Sven Burgdorf, Nicholas A Melosh, Evan W Newell, Florent Ginhoux, Andreas Schlitzer, Joachim L Schultze
ABSTRACT

Human in vitro generated monocyte-derived dendritic cells (moDCs) and macrophages are used clinically, e.g., to induce immunity against cancer. However, their physiological counterparts, ontogeny, transcriptional regulation, and heterogeneity remains largely unknown, hampering their clinical use. High-dimensional techniques were used to elucidate transcriptional, phenotypic, and functional differences between human in vivo and in vitro generated mononuclear phagocytes to facilitate their full potential in the clinic. We demonstrate that monocytes differentiated by macrophage colony-stimulating factor (M-CSF) or granulocyte macrophage colony-stimulating factor (GM-CSF) resembled in vivo inflammatory macrophages, while moDCs resembled in vivo inflammatory DCs. Moreover, differentiated monocytes presented with profound transcriptomic, phenotypic, and functional differences. Monocytes integrated GM-CSF and IL-4 stimulation combinatorically and temporally, resulting in a mode- and time-dependent differentiation relying on NCOR2. Finally, moDCs are phenotypically heterogeneous and therefore necessitate the use of high-dimensional phenotyping to open new possibilities for better clinical tailoring of these cellular therapies.

MATERIALS
Product Number
Brand
Product Description

Millipore
Lactobacillus bulgaricus Agar (Base), NutriSelect® Plus, suitable for microbiology
Sigma-Aldrich
Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone, ≥98% (TLC), powder
SAFC
L-Glutamine Solution 200 mM, 29.23 mg/mL in saline, solution, suitable for cell culture
Sigma-Aldrich
Anti-Tubulin Antibody, clone YL1/2, clone YL1/2, Chemicon®, from rat
Sigma-Aldrich
Oligomycin A, ≥99% (HPLC)
Sigma-Aldrich
Anti-Actin Antibody, clone C4, ascites fluid, clone C4, Chemicon®
Sigma-Aldrich
MISSION® esiRNA, targeting human NCOR2