Tumor Necrosis Factor Alpha Enhances the Adenoviral Transduction of CD34 + Hematopoietic Progenitor Cells

Abstract

The purpose of this study was to improve the transduction efficiency of adenoviral vectors (Ad) in human CD34 + hematopoietic progenitor cells. CD34 + cells from cord blood or mobilized peripheral blood were incubated with tumor necrosis factor-alpha (TNF-α). After removal of free TNF-α, the cells were infected with an Ad encoding green fluorescent protein (GFP). One day later, viable cells were counted and analyzed for GFP and CD34 by flow cytometry. To visualize vectoral trafficking, CD34 + cells were incubated with fluorophore-conjugated Ad. Plating efficiencies of hematopoietic progenitors before and after transduction were evaluated by methylcellulose assays. Pretreatment with TNF-α increased the transduction efficiency more than twofold (39.2% versus 15.5%) in a dose-dependent manner and strongly improved the survival of GFP-positive CD34 + cells. Time course experiments showed that TNF-α incubation times as short as 10 minutes were still effective. Neutralizing antibodies to TNF receptor II and RGD peptides diminished the TNF-α-dependent increase in transduction efficiency. No TNF-α-dependent increase in adenoviral receptors (coxsackie-adenovirus receptor, α v β 3 -integrin) occurred. Analysis of viral binding demonstrated a significantly higher incidence of local concentrations of Ad along the cell surface (caps) in virus-positive cells of the TNF-α-treated group. Plating efficiency, especially the formation of granulocyte-macrophage colony forming units, was enhanced by TNF-α pretreatment. We conclude that brief incubation with TNF-α before addition of the Ad significantly increased the Ad transduction efficiency in CD34 + cells, and improved post-transduction survival of progenitors of the granulocyte-macrophage lineage. This finding correlates with increased Ad capping at the cell surface and suggests an alteration of Ad trafficking.