Tumor necrosis factor alpha enhances the adenoviral transduction of CD34+ hematopoietic progenitor cells Journal Article
| Authors: | Moldenhauer, A.; Shieh, J.H.; Pruss, A.; Salama, A.; Moore, M. A. S. |
| Article Title: | 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, αvβ3-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. |
| Keywords: | controlled study; nonhuman; flow cytometry; cell viability; cell survival; cd34 antigen; granulocyte macrophage colony stimulating factor; tumor necrosis factor receptor 2; green fluorescent protein; cell lineage; genetic transduction; genetic vectors; transduction, genetic; fetal blood; tumor necrosis factor alpha; tumor necrosis factor-alpha; hematopoietic stem cells; umbilical cord blood; hematopoietic stem cell; adenovirus vector; oligopeptides; antigens, cd34; arginylglycylaspartic acid; adenoviridae; receptors, tumor necrosis factor; colony forming unit; cell surface; adenovirus; transduction; neutralizing antibody; vitronectin receptor; capping; tnf-α; humans; human; article; methylcellulose; cd34+ cells; coxsackie virus; coxsackievirus |
| Journal Title: | Stem Cells |
| Volume: | 22 |
| Issue: | 3 |
| ISSN: | 1066-5099 |
| Publisher: | AlphaMed Press |
| Date Published: | 2004-05-01 |
| Start Page: | 283 |
| End Page: | 291 |
| Language: | English |
| PROVIDER: | scopus |
| PUBMED: | 15153606 |
| DOI: | 10.1634/stemcells.22-3-283 |
| DOI/URL: | |
| Notes: | Stem Cells -- Cited By (since 1996):9 -- Export Date: 16 June 2014 -- CODEN: STCEE C2 - 15153606 -- Source: Scopus |
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