Efficient infection of primitive hematopoietic stem cells by modified adenovirus

Abstract

Almost all studies of adenoviral vector-mediated gene transfer have made use of the adenovirus type 5 (Ad5). Unfortunately, Ad5 has been ineffective at infecting hematopoietic progenitor cells (HPC). Chimeric Ad5/F35 vectors that have been engineered to substitute the shorter-shafted fiber protein from Ad35 can efficiently infect committed hematopoietic cells and we now show highly effective gene transfer to primitive progenitor subsets. An Ad5GFP and Ad5/F35GFP vector was added to CD34 + and CD34 − lineage − (lin − ) HPC. Only 5-20% of CD34 + and CD34 − lin − cells expressed GFP after Ad5 exposure. In contrast, with the Ad5/F35 vector, 30-70% of the CD34 + , 50-70% of the CD34 − lin − and up to 60% of the CD38 − HPC expressed GFP and there was little evident cellular toxicity. Because of these improved results, we

Osteoinduction by ex vivo adenovirus-mediated BMP2 delivery is independent of cell type

“…One candidate target cell for Ad-BMP2 could be nucleated cells of the peripheral blood. We 24 and others 25 have demonstrated the suitability of Ad5F35 vectors, unlike Ad5 vectors, to transduce efficiently human hematopoietic cells.…”

In vivo transduction of primitive mobilized hematopoietic stem cells after intravenous injection of integrating adenovirus vectors

“…Both others and we have shown that Ad vectors containing the Ad35 fiber or fiber knob (Ad5/35) efficiently transduce human and non-human primate HSPCs in vitro. [7][8][9][10] Integration site analysis Genomic DNA was isolated from GFP 1 hCD46tg colonies using the DNeasy Blood and Tissue Kit (Qiagen, Valencia, CA). These CFU colonies were obtained from Lin 2 /GFP 1 BM cells of in vivo transduced hCD46tg animals.…”

Tightly regulated gene expression in human hematopoietic stem cells after transduction with helper-dependent Ad5/35 vectors

“…Previous studies showed that Ad5/35 vectors efficiently infect human CD34 + cells, particularly subsets with potential stem cell activity. For example, at an MOI of 10pfu/cell, Ad5/35 transduced the majority of CD34 + /c-Kit + [4], CD34 + /CD38 -, CD34 -/Lin -, and Hoechst 33342 dye-negative side population (SP) cells [6]. Furthermore, Ad5/35 vectors allowed for efficient gene transfer into non-dividing CD34 + cells [7], which is important as most of the repopulating HSCs are quiescent.…”