Figure 5

Functional analyses of the ex vivo expanded DCPs. (A) Analysis of antigen capture function of the ex vivo derived DCPs. Antigen capture was demonstrated using dextran-FITC or OVA-FITC particle internalization followed by flow cytometry analysis. Examples of FITC-positive control PBMC-derived DCs are shown at top; antigen capture was detected at 37°C but not at 4°C. (B) Analysis of antigen-specific T cell stimulation function. DCs were transduced with LVs encoding a control truncated NGFR (tNGFR) protein or BMLF protein of EBV, and incubated with autologous T cells (from the same HLA-A*0201 donor) for 10-12 days. The BMLF-specific A*0201 TCR bearing T cells were detected using a PE-conjugated MHC-peptide pentamer by flow cytometry (left panel). Antigen-specific effector function was analyzed based on intracellular expression of IFN-γ as described in Materials and Methods.