Figure 2.

Ad-FBAP-GFP transduction of primary human monocyte-derived DC can be enhanced by targeting to alternative receptors. Primary human monocyte-derived DC were used on day 7 following isolation of PBMC. Cells were incubated with unmodified rAd vectors (Ad-FBAP-GFP), or protein/peptide-NA-virus complexes of interest, at a multiplicity of infection of 2000 viral genome units (vg) per cell, and then GFP expression was analyzed 24-28 hours thereafter, using flow cytometry. (A) Analysis of the effect of targeting virus to DC-SIGN: GFP expression data are shown for virus that was conjugated to a DC-SIGN specific monoclonal antibody (dark bars) versus virus that was targeted using an irrelevant antibody of the same isotype (open bars). (B) Analysis of the effect of targeting virus to α_vβ₃ integrin. GFP expression data are shown for cultures which were transduced with virus that had been left unmodified (Ad-FBAP-GFP) or conjugated to the following peptides: (i) a low-affinity and relatively non-selective α_vβ₃ integrin-binding (FnFn10), (ii) a high- affinity and highly selective α_vβ₃ integrin-binding peptide (3JCLI4). (C) Analysis of the effect of targeting virus to ChemR23: GFP expression data are shown for virus that was conjugated to (i) a mutated derivative of a ChemR23-binding peptide, which contains an alanine substitution in a key residue (chemerin FAAS), or (ii) the wild-type version of this same peptide, which binds to ChemR23 with nanomolar affinity (chemerin FAFS). (A, B,C) Data represent mean values from 3 experiments; bars denote the standard error of the mean. Statistically significant differences are indicated as p values above the compared groups (t test).