Fig. 1
Adoptive transfer of Ag and adjuvant-treated natural mouse cDC1s promotes CD8+ T cell immunity. a Quantitative PCR analyses of mRNA levels of Ifnb1 (IFNβ protein), Il12b (IL12-p40 protein), CD40 and CCR7 relative to Actb (β-actin protein) of spleen cDC1s freshly isolated from B16-FLT3L tumor-bearing mice and treated with 20 μg/ml poly I:C (InvivoGen) for 1 h ex vivo, n = 4 spleen cDC1 preparations. *P < 0.05 by Ratio paired Student’s t test. Ct: Cycle threshold, −ΔCt = −(Ct [gene of interest] - Ct [Actb internal control gene]). b Schematic representation of experimental setup for data shown in c-f. Spleen cDC1s from B16-FLT3L tumor-bearing mice were cultured with 20 μg/ml poly I:C and/or 20 μg/ml soluble OVA protein for 1 h, washed and 2 × 105 cDC1s injected intravenously into CD45.2+ recipient mice that had been adoptively transferred with 1-2 × 105 CellTrace-Violet (CV)-labelled CD45.1+ OT-I CD8+ T cells one-day prior. c Representative flow cytometric analysis of OT-I T cell proliferation via CV-dilution in the spleen at day 5, gated on CD45.1+ CD8+ OT-I cells. d-f Flow cytometric quantification of d total OT-I cell number, e OT-I frequency in CD8+ cells and f number of IFNγ-producing OT-I cells after re-stimulation with OVA257–264 peptide in the spleen at day 5 post-cDC1 injection. One representative of 2 independent experiments (n = 3–4 mice/group/experiment) is shown. *P < 0.05, **P < 0.01 by Student’s t test