Rapid induction of β-galactosidase transgene expression after adenovirus infection of FR901228-treated cells.

Frozen PBMCs and CD34⁺ PBSCs from different donors were thawed and incubated in medium overnight. FR901228 was added to the treated cells, and all cells were incubated for 24 hours. (A) RT-PCR analysis of CAR RNA in FR901228-treated PBMCs. PBMCs were treated without (−) or with the indicated concentration of FR901228. Samples were prepared as described in the legend to Figure 1. (B) Quantitation of β-galactosidase– positive cells after adenovirus infection. Untreated cells or cells treated with 0.1 ng/mL FR901228 were infected with the indicated MOI of ADCMVβgal for 1 hour in the absence of serum. Serum was added to the medium. Cells were incubated for 24 hours and were analyzed for β-galactosidase activity as described in the legend to Figure 2. The viability of the PBMCs, as determined by trypan blue exclusion, was at least 94%. The viability of CD34⁺PBSCs, as determined by propidium iodide exclusion, was 95%. In addition, 92% of these cells showed expression of the CD34 epitope at the conclusion of the experiment.