*2.5. Generation of Human Monocyte-Derived Dendritic Cells (moDCs)*

PBMCs were isolated from buffy coats by density gradient centrifugation using histopaque-1077 (Sigma, St. Louis, MO, USA). The buffy coats from donors were kindly provided by the SALK (Salzburger Landesklinikum). The monocytes were then separated from the PBMCs using the adherence method. The adherent monocytes were then cultured in dendritic cell medium containing RPMI (Sigma), 10% heat-inactivated FCS (Biowest, Nuaillé, France), 1% 2 mM L-glutamine (Sigma), 1% Pen–Strep (Sigma), 50 mM 2-mercaptoethanol (Sigma) supplemented with 50 ng/mL granulocyte macrophage colonystimulating factor (GM-CSF) (Life Technologies, Carlsbad, CA, USA) and interleukin 4 (IL-4) (Life Technologies, Carlsbad, CA, USA) for 6 days. The cells were refed with 100 ng/mL of GM-CSF and IL-4 on day 3. The protocol for generation of moDCs is described in detail by Posselt et al. [29].
