Because of a printing error,
Figures 1, 2, 3, 4, and 5 of "Targeted adeno-associated virus vector transduction of nonpermissive cells mediated by a bispecific F(ab ´γ) 2 antibody," by Jeffrey S. Bartlett, Jurgen Kleinschmidt, Richard C. Boucher, and R. Jude Samulski, were printed incorrectly. The correct figures and captions now appear. Figure 1: Quantification of β-galactosidase expression in cell lines transduced with recombinant AAVβgal vector.
AAVβgal was incubated with cells for 1 h at 37°C. The indicated samples then received adenovirus (dl309) for 1 h. Activities of mock infections that received no vector were subtracted for each cell line. Results represent the average of duplicate samples.
Figure 2: AAV binding to different cells.
The indicated cells were incubated with [
35S]methionine-[ 35S]cysteine-labeled virus for 1 h at 4°C and washed, and cell-associated activity was determined in a scintillation counter. Open columns (□) represent total binding. Closed columns (▪) indicate binding in the presence of a 30-fold excess of nonlabeled virus. Experiments were performed in triplicate and bars indicate the standard error of the mean. Figure 3: Fluorescence-activated cell sorting (FACS) analysis of α IIbβ 3 integrin expression.
Cells were treated with an excess amount of AP-2 Mab or control mouse IgG, washed and stained with phycoerythrin-conjugated polyclonal donkey antimouse F(ab´)
2. FACS profiles of control IgG-stained cells (– – –), and AP-2–stained cells (——) are shown. Figure 4: Targeted transduction of cells mediated by bispecific antibody.
AAVβgal, or the AAVβgal-bispecific Ab complex was incubated with the various cells for 1 h at 37°C. Vector–bispecific antibody complexes were preformed for 1 h prior to addition to cells. When indicated, cells were pretreated for 1 h at 37°C with 25 ng/ml AP-2, or AK2 antibody. Following incubation with vector, the cells were washed and grown for 24 h at 37°C, and β-galactosidase activity was then determined as described in the Experimental Protocol. Activities of mock infections that received no vector are shown for each cell line. Results represent the average of triplicate samples from three to five experiments, and bars indicate the standard error of the mean.
Figure 5: Binding of radiolabeled AAV to human megakaryocytic leukemia cell lines in the presence of bispecific F(ab ´γ) 2.
Radiolabeled AAV was preincubated with bispecific A20AP2 F(ab ´γ)
2 then added to DAMI, MO7e, or HeLa cells in the presence (□) or absence (▪) of competing AP-2 antibody. Cell-associated radioactivity, corrected for specific binding by subtracting values obtained in the presence of a 50-fold excess of unlabeled virus, was determined in a scintillation counter. Experiments were performed in triplicate, and bars indicate the standard error of the mean.