title: Retroviral transfer of the nlsLacZ gene into human CD34+ cell populations and into TF-1 cells: future prospects in gene therapy.
abstract: Few data are available concerning behavior of reimplanted human hematopoietic cells after autologous stem cell transplantation. This paper reports the possibility to transfer gene markers coding for beta-galactosidase (beta-Gal) activity by retroviral vectors into a human leukemic growth factor-dependent cell line, TF-1, and into human hematopoietic progenitors isolated from peripheral blood or bone marrow. Using various combinations of retroviral vectors and packaging cell lines, we demonstrated high expression of a bacterial beta-Gal activity induced by the LacZ gene, the nlsLacZ gene, or the Sh-ble/LacZ gene, in human hematopoietic cells. The expression of the nlsLacZ construct was stable until the end of the culture in infected CD34+ cell-enriched cell populations, and a slow decrease of transgene expression was observed in a transduced TF-1 cell population during a 1-year long-term culture. Data obtained with the nlsLacZ gene demonstrate that both retroviral transfer and corresponding gene expression were not found to modify the pattern of cell proliferation and differentiation. These results open interesting prospectives for the use of the nlsLacZ gene to mark and follow the fate of progenitor cells isolated from patients with cancers prior to reimplantation.
authors.affiliation: Institut Paoli-Calmettes and INSERM U119, Marseille, France.
authors.forename: C
authors.initials: C
authors.lastname: Bagnis
authors.forename: G
authors.initials: G
authors.lastname: Gravis
authors.forename: A M
authors.initials: AM
authors.lastname: Imbert
authors.forename: D
authors.initials: D
authors.lastname: Herrera
authors.forename: T
authors.initials: T
authors.lastname: Allario
authors.forename: R
authors.initials: R
authors.lastname: Galindo
authors.forename: M
authors.initials: M
authors.lastname: Lopez
authors.forename: C
authors.initials: C
authors.lastname: Pavon
authors.forename: C
authors.initials: C
authors.lastname: Sempere
authors.forename: P
authors.initials: P
authors.lastname: Mannoni
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000032:analysis; D018952:Antigens, CD34; D002460:Cell Line; D005544:Forecasting; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D016298:Hematopoietic Cell Growth Factors / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000378:metabolism; D006801:Humans; D007938:Leukemia / Q000473:pathology; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D012190:Retroviridae / Q000235:genetics*; D013997:Time Factors; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000187:drug effects; D001616:beta-Galactosidase / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D015703:Antigens, CD; D018952:Antigens, CD34; D016298:Hematopoietic Cell Growth Factors; D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase