Transduction efficay of an Ad vector at various MOIs and surface marker expression of adenovirus-infected DCs. (A) Recombinant AdGFP was used to transduce day 7 immature DCs, which were then cultured for 48 h in the presence of GM-CSF and IL-4. On day 9, flow cytometric analysis of GFP expression by the Ad-GFP-DCs was carried out. Typically, > 74% of cells were GFP + at a MOI of 200. (B) IL-2 and/or AFP expression by gene-modified DCs is shown. PCR products of β-actin, IL-2 and AFP were visualized by electrophoresis in a 2% agarose gel containing 0.5 μg/ml ethidium bromide. Lane 1 Marker, lane 2 IL-2-DC, lane 3 IL-2/AFP-DC, lane 4 AFP-DC, lane 5 GFP-DC, lane 6 DC group. Ad, adenovirus; MOI, multiplicity of infection; DCs, dendritic cells; GFP, green fluorescent protein; GM-CSF, granulocyte macrophage-colony stimulating factor; IL-2, inter-leukin-2; AFP, α-fetoprotein; PCR, polymerase chain reaction.
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