Critical factors influencing stable transduction of human CD34(+) cells with HIV-1-derived lentiviral vectors.

:Lentiviral vectors have been proposed as a more efficient alternative to Moloney murine leukemia virus-based retroviral vectors for transduction of human hematopoietic progenitors and stem cells. These studies were designed to evaluate the conditions that influence transduction frequency of CD34(+) progenitors, with the goal of optimizing efficiency of stable gene transfer with lentiviral vectors. CD34(+) human cord blood cells and 293 cells were transduced with a human immunodeficiency virus (HIV)-1 derived lentiviral vector pseudotyped with vesicular stomatitis virus glycoprotein and carrying an internal human cytomegalovirus promoter driving enhanced green fluorescent protein (eGFP) expression. Using fluorescence-activated cell sorting analysis of eGFP, we observed pseudotransduction beginning at the time of vector addition and lasting up to 24 h in CD34(+) cells and up to 72 h in 293 cells. Integrase-defective lentiviral vector caused transient eGFP expression for up to 10 days in CD34(+) cells and for up to 14 days in 293 cells. Protamine sulfate conferred no increase in transduction efficiency of CD34(+) cells on fibronectin-coated plates. Transduction frequency was related directly to vector concentration and not to multiplicity of infection across the ranges tested. First- and second-generation lentiviral vectors transduced CD34(+) cells equally, demonstrating a lack of dependence on HIV-1 accessory proteins. These findings will be useful for the optimal utilization of this new class of vectors for transduction of human hematopoietic stem cells.

Haas DL ，Case SS ，Crooks GM ，Kohn DB ... -  《MOLECULAR THERAPY》

Hybrid HIV/MSCV LTR enhances transgene expression of lentiviral vectors in human CD34(+) hematopoietic cells.

Choi JK ，Hoang N ，Vilardi AM ，Conrad P ，Emerson SG ，Gewirtz AM ... -  《STEM CELLS》

High levels of transgene expression following transduction of long-term NOD/SCID-repopulating human cells with a modified lentiviral vector.

Gao Z ，Golob J ，Tanavde VM ，Civin CI ，Hawley RG ，Cheng L ... -  《STEM CELLS》

Transduction of human CD34+ CD38- bone marrow and cord blood-derived SCID-repopulating cells with third-generation lentiviral vectors.

Guenechea G ，Gan OI ，Inamitsu T ，Dorrell C ，Pereira DS ，Kelly M ，Naldini L ，Dick JE ... -  《MOLECULAR THERAPY》

Expression from second-generation feline immunodeficiency virus vectors is impaired in human hematopoietic cells.

Price MA ，Case SS ，Carbonaro DA ，Yu XJ ，Petersen D ，Sabo KM ，Curran MA ，Engel BC ，Margarian H ，Abkowitz JL ，Nolan GP ，Kohn DB ，Crooks GM ... -  《MOLECULAR THERAPY》