title: Transduction of human primitive repopulating hematopoietic cells with lentiviral vectors pseudotyped with various envelope proteins.
abstract: Lentiviral vectors are useful for transducing primitive hematopoietic cells. We examined four envelope proteins for their ability to mediate lentiviral transduction of mobilized human CD34(+) peripheral blood cells. Lentiviral particles encoding green fluorescent protein (GFP) were pseudotyped with the vesicular stomatitis virus envelope glycoprotein (VSV-G), the amphotropic (AMPHO) murine leukemia virus envelope protein, the endogenous feline leukemia viral envelope protein or the feline leukemia virus type C envelope protein. Because the relative amount of genome RNA per ml was similar for each pseudotype, we transduced CD34(+) cells with a fixed volume of each vector preparation. Following an overnight transduction, CD34(+) cells were transplanted into immunodeficient mice which were sacrificed 12 weeks later. The average percentages of engrafted human CD45(+) cells in total bone marrow were comparable to that of the control, mock-transduced group (37-45%). Lenti-particles pseudotyped with the VSV-G envelope protein transduced engrafting cells two- to tenfold better than particles pseudotyped with any of the gamma-retroviral envelope proteins. There was no correlation between receptor mRNA levels for the gamma-retroviral vectors and transduction efficiency of primitive hematopoietic cells. These results support the use of the VSV-G envelope protein for the development of lentiviral producer cell lines for manufacture of clinical-grade vector.
authors.affiliation: Department of Experimental Hematology, St Jude Children's Research Hospital, Memphis, Tennessee 38105, USA.
authors.forename: Yoon-Sang
authors.initials: YS
authors.lastname: Kim
authors.forename: Matthew M
authors.initials: MM
authors.lastname: Wielgosz
authors.forename: Phillip
authors.initials: P
authors.lastname: Hargrove
authors.forename: Steven
authors.initials: S
authors.lastname: Kepes
authors.forename: John
authors.initials: J
authors.lastname: Gray
authors.forename: Derek A
authors.initials: DA
authors.lastname: Persons
authors.forename: Arthur W
authors.initials: AW
authors.lastname: Nienhuis
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D002460:Cell Line; D005822:Genetic Vectors / Q000235:genetics*; D006367:HeLa Cells; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D005256:Leukemia Virus, Feline / Q000378:metabolism; D009052:Leukemia Virus, Murine / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics; D051379:Mice; D014161:Transduction, Genetic / Q000379:methods*; D014759:Viral Envelope Proteins / Q000235:genetics
chemical_list: D018952:Antigens, CD34; C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins