Adenovirus-mediated expresssion of the murine ecotropic receptor facilitates transduction of human hematopoietic cells with an ecotropic retroviral vector.

:One factor limiting the ability to modify human repopulating hematopoietic cells genetically with retroviral vectors is the relatively low expression of the cognate viral receptor. We have tested sequential transduction of human hematopoietic cells with an adenoviral vector encoding the ecotropic retroviral receptor followed by transduction with an ecotropic retroviral vector. Adenoviral transduction of K562 erythroleukemia cells was highly efficiently with >95% of cells expressing the ecotropic receptor at a multiplicity of infection (MOI) of 103with a correspondingly high transduction with a retroviral vector. Ecotropic receptor expression in CD34+ cells following transduction with adenoviral vectors was increased by at least two-fold (from 20 to 48%) by replacing the RSV promoter with the CMV E1a promoter, resulting in a parallel increase in retroviral transduction efficiency. Replacing the head portion of the fiber protein in conventional adenoviral vectors (serotype 5) with the corresponding portion from an adenoviral 3 serotype resulted in ecotropic receptor expression in 60% of CD34+ cells at an MOI of 104 and a retroviral transduction of 60% of hematopoietic clonogenic progenitors. The sequential transduction strategy also resulted in efficient transduction of the primitive CD34+CD38- subset suggesting that it may hold promise for genetic modification of human hematopoietic stem cells.

Nathwani AC ，Persons DA ，Stevenson SC ，Frare P ，McClelland A ，Nienhuis AW ，Vanin EF ... -  《GENE THERAPY》

Efficient gene transfer into human cord blood CD34+ cells and the CD34+CD38- subset using highly purified recombinant adeno-associated viral vector preparations that are free of helper virus and wild-type AAV.

Nathwani AC ，Hanawa H ，Vandergriff J ，Kelly P ，Vanin EF ，Nienhuis AW ... -  《GENE THERAPY》

Adenovirus facilitated infection of human cells with ecotropic retrovirus.

Scott-Taylor TH ，Gallardo HF ，Gänsbacher B ，Sadelain M ... -  《GENE THERAPY》

Optimization of adenovirus serotype 35 vectors for efficient transduction in human hematopoietic progenitors: comparison of promoter activities.

Sakurai F ，Kawabata K ，Yamaguchi T ，Hayakawa T ，Mizuguchi H ... -  《GENE THERAPY》

Adenoviral vector design for high-level transgene expression in primitive human hematopoietic progenitors.

Fan X ，Brun A ，Karlsson S 《GENE THERAPY》