title: Efficient infection of a human T-cell line and of human primary peripheral blood leukocytes with a pseudotyped retrovirus vector.
abstract: Peripheral blood lymphocytes (PBLs) are an important target for gene transfer studies aimed at human gene therapy. However, no reproducibly efficient methods are currently available to transfer foreign, potentially therapeutic genes into these cells. While vectors derived from murine retroviruses have been the most widely used system, their low infection efficiency in lymphocytes has required prolonged in vitro culturing and selection after infection to obtain useful numbers of genetically modified cells. We previously reported that retroviral vectors pseudotyped with vesicular stomatitis G glycoprotein (VSV-G) envelope can infect a wide variety of cell types and can be concentrated to titers of greater than 10(9) infectious units/ml. In this present study, we examined the ability of amphotropic and pseudotyped vectors expressing a murine cell surface protein, B7-1, to infect the human T-cell line Jurkat or human blood lymphocytes. Limiting dilution analysis of transduced Jurkat cells demonstrated that the pseudotyped vector is significantly more efficient in infecting T cells than an amphotropic vector used at the same multiplicity of infection (moi). To identify the transduction efficiency on PBLs, we examined the levels of cell surface expression of the B7-1 surface marker 48 to 72 hr after infection. The transduction efficiency of PBLs with the pseudotyped vector increased linearly with increasing moi to a maximum of approximately 16-32% at an moi of 40. This relatively high efficiency of infection of a T-cell line and of blood lymphocytes with VSV-G pseudotyped virus demonstrates that such modified pseudotyped retrovirus vectors may be useful reagents for studies of gene therapy for a variety of genetic or neoplastic disorders.
authors.affiliation: Department of Pediatrics, University of California School of Medicine at San Diego, La Jolla 92093-0634, USA.
authors.forename: S
authors.initials: S
authors.lastname: Sharma
authors.forename: M
authors.initials: M
authors.lastname: Cantwell
authors.forename: T J
authors.initials: TJ
authors.lastname: Kipps
authors.forename: T
authors.initials: T
authors.lastname: Friedmann
mesh_terms: D000818:Animals; D018122:B7-1 Antigen / Q000096:biosynthesis*; D015496:CD4-Positive T-Lymphocytes / Q000502:physiology; D002460:Cell Line; D002478:Cells, Cultured; D005434:Flow Cytometry; D015870:Gene Expression; D005819:Genetic Markers; D005822:Genetic Vectors; D006801:Humans; D007376:Interleukin-2 / Q000494:pharmacology; D008214:Lymphocytes / Q000187:drug effects / Q000502:physiology*; D051379:Mice; D011994:Recombinant Proteins / Q000494:pharmacology; D012190:Retroviridae; D013601:T-Lymphocytes / Q000502:physiology*; D014162:Transfection / Q000379:methods*; D014407:Tumor Cells, Cultured
chemical_list: D018122:B7-1 Antigen; D005819:Genetic Markers; D007376:Interleukin-2; D011994:Recombinant Proteins