title: Engraftment of NOD/SCID mice with human CD34(+) cells transduced by concentrated oncoretroviral vector particles pseudotyped with the feline endogenous retrovirus (RD114) envelope protein.
abstract: Oncoretrovirus vectors pseudotyped with the feline endogenous retrovirus (RD114) envelope protein produced by the FLYRD18 packaging cell line have previously been shown to transduce human hematopoietic progenitor cells with a greater efficiency than similar amphotropic envelope-pseudotyped vectors. In this report, we describe the production and efficient concentration of RD114-pseudotyped murine leukemia virus (MLV)-based vectors. Following a single round of centrifugation, vector supernatants were concentrated approximately 200-fold with a 50 to 70% yield. Concentrated vector stocks transduced prestimulated human CD34(+) (hCD34(+)) cells with approximately 69% efficiency (n = 7, standard deviation = 4.4%) using a single addition of vector at a low multiplicity of infection (MOI = 5). Introduction of transduced hCD34(+) cells into irradiated NOD/SCID recipients resulted in multilineage engraftment with long-term transgene expression. These data demonstrate that RD114-pseudotyped MLV-based vectors can be efficiently concentrated to high titers and that hCD34(+) cells transduced with concentrated vector stocks retain in vivo repopulating potential. These results highlight the potential of RD114-pseudotyped oncoretrovirus vectors for future clinical implementation in hematopoietic stem cell gene transfer.
authors.affiliation: Division of Infectious Diseases, Department of Internal Medicine, University of Texas Southwestern Medical Center at Dallas, 75390-9113, USA.
authors.forename: J
authors.initials: J
authors.lastname: Gatlin
authors.forename: M W
authors.initials: MW
authors.lastname: Melkus
authors.forename: A
authors.initials: A
authors.lastname: Padgett
authors.forename: P F
authors.initials: PF
authors.lastname: Kelly
authors.forename: J V
authors.initials: JV
authors.lastname: Garcia
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000276:immunology; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000276:immunology; D006801:Humans; D016089:Immunodeficiency Virus, Feline / Q000737:chemistry*; D009052:Leukemia Virus, Murine / Q000235:genetics*; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D014759:Viral Envelope Proteins / Q000235:genetics*
chemical_list: D018952:Antigens, CD34; D014759:Viral Envelope Proteins