Fig. 1
Defective cross-presentation by DC in cancer. a–d Proliferation of CD8+ T cells measured at 72 h by CFSE dilution. DCs were obtained from dLNs of naïve or TB mice 3 days after immunization with HPV DNA (a–c) or OVA-LV vaccines (d). CD8+ T cells were isolated from mice immunized with HPV E6/E7 vaccine (a–c) or from OT1 transgenic mice (d). a Antigen-specific CD8+ T cells; b allogeneic T cells; c antigen-specific CD8+ T cell proliferation after stimulation with DCs isolated from dLNs of naïve or TB mice and then loaded ex vivo with HPV-derived peptides. d proliferation of OT1 CD8+ T cells (left panel) and allogenic T cells (right panel) after stimulation with DCs isolated from LN of mice immunized with OVA-LV. Each group included three mice. e proliferation of OT1 CD8+ T cells after stimulation with DC generated with GM-CSF, treated with TES and loaded with OVA protein (left panel) or short peptides (SIINFKEL) (right panel). Typical example of 7 experiments is shown. f, h Expression of pMHC and MHC class I (H2Kb) by DC generated with GM-CSF (f) or FLT3 ligand (h) treated with TES for 48 h and then loaded with OVA-derived long or short peptides. Typical example of seven experiments is shown. g, i Proliferation of OT1 CD8+ T cells after stimulation with DC generated with GM-CSF (g) or FLT3 ligand (i) (left panels: DCs loaded with long peptide, right panels: DCs loaded with short peptide). Typical example of seven experiments is shown. j Proliferation of OT1 CD8+ T cells after stimulation with DC treated with indicated supernatants for 48 h and loaded with OVA-derived long peptides. k, l DCs generated in vitro from BM progenitors with GM-CSF and FLT3L were treated with TES for 48 h. k Typical phenotype of cells. l Proliferation of antigen-specific CD8+ T cells after stimulation with sorted CD103+DCs. Cumulative results of three experiments are shown. e–l Proliferation was measured by 3[H]-thymidine uptake in triplicates. In all experiments mean and SD are shown *p < 0.05, **p < 0.01 in unpaired two-tailed Student's t test between compared groups
