Figure 7
Figure 7. Uptake of CMV-infected human foreskin fibroblasts (HFF) and cross-presentation of CMV-specific peptides. Phagocytosis of CMV strain AD169 infected (CMV+) or uninfected (CMV−) HFFs (HLA-A2 negative) and subsequent cross-presentation of CMV-derived peptides on HLA-A2 molecules was analyzed in IFNγ-ELIspot assays. (A) The generated CTL line raised against a HLA-A2–binding CMV pp65-derived peptide specifically recognized autologous MDCs loaded with the corresponding peptide but not an irrelevant HIV-derived peptide. (B) Infected or uninfected HFFs were cocultured with immature MDCs in the presence or absence of zymosan or the hDectin-1b ECD, respectively (representative experiment). (C) Uptake of infected HFFs by MDCs that were matured using different TLR ligands or were left in an immature state (GM/IL-4) (representative experiment). (D) Addition of the hDectin-1–specific (1→3)-β-D-glucan curdlan to the coculture of MDCs and infected HFFs led to a reduction of IFNγ production by the CMV-specific CTL line, while addition of an irrelevant protein (DHFR; 10.0 μg/mL) or the buffer control did not (representative experiment). (E) The diagram represents mean (± SD) of 3 independent experiments using different donors. *P < .05 (paired t test).

Uptake of CMV-infected human foreskin fibroblasts (HFF) and cross-presentation of CMV-specific peptides. Phagocytosis of CMV strain AD169 infected (CMV+) or uninfected (CMV) HFFs (HLA-A2 negative) and subsequent cross-presentation of CMV-derived peptides on HLA-A2 molecules was analyzed in IFNγ-ELIspot assays. (A) The generated CTL line raised against a HLA-A2–binding CMV pp65-derived peptide specifically recognized autologous MDCs loaded with the corresponding peptide but not an irrelevant HIV-derived peptide. (B) Infected or uninfected HFFs were cocultured with immature MDCs in the presence or absence of zymosan or the hDectin-1b ECD, respectively (representative experiment). (C) Uptake of infected HFFs by MDCs that were matured using different TLR ligands or were left in an immature state (GM/IL-4) (representative experiment). (D) Addition of the hDectin-1–specific (1→3)-β-D-glucan curdlan to the coculture of MDCs and infected HFFs led to a reduction of IFNγ production by the CMV-specific CTL line, while addition of an irrelevant protein (DHFR; 10.0 μg/mL) or the buffer control did not (representative experiment). (E) The diagram represents mean (± SD) of 3 independent experiments using different donors. *P < .05 (paired t test).

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