title: Gene delivery to human B-precursor acute lymphoblastic leukemia cells.
abstract: Autologous leukemia cells engineered to express immune-stimulating molecules may be used to elicit antileukemia immune responses. Gene delivery to human B-precursor acute lymphoblastic leukemia (ALL) cells was investigated using the enhanced green fluorescent protein (EGFP) as a reporter gene, measured by flow cytometry. Transfection of the Nalm-6 and Reh B-precursor ALL leukemia cell lines with an expression plasmid was investigated using lipofection, electroporation, and a polycationic compound. Only the liposomal compound Cellfectin showed significant gene transfer (3.9% to 12% for Nalm-6 cells and 3.1% to 5% for Reh cells). Transduction with gibbon-ape leukemia virus pseudotyped Moloney murine leukemia virus (MoMuLV)-based retrovirus vectors was investigated in various settings. Cocultivation of ALL cell lines with packaging cell lines showed the highest transduction efficiency for retroviral gene transfer (40.1% to 87.5% for Nalm-6 cells and 0.3% to 9% for Reh cells), followed by transduction with viral supernatant on the recombinant fibronectin fragment CH-296 (13% to 35.5% for Nalm-6 cells and 0.4% to 6% Reh cells), transduction on human bone marrow stroma monolayers (3.2% to 13.3% for Nalm-6 cells and 0% to 0.2% Reh cells), and in suspension with protamine sulfate (0.7% to 3.1% for Nalm-6 cells and 0% for Reh cells). Transduction of both Nalm-6 and Reh cells with human immunodeficiency virus-type 1 (HIV-1)-based lentiviral vectors pseudotyped with the vesicular stomatitis virus-G envelope produced the best gene transfer efficiency, transducing greater than 90% of both cell lines. Gene delivery into primary human B-precursor ALL cells from patients was then investigated using MoMuLV-based retrovirus vectors and HIV-1-based lentivirus vectors. Both vectors transduced the primary B-precursor ALL cells with high efficiencies. These studies may be applied for investigating gene delivery into primary human B-precursor ALL cells to be used for immunotherapy.
authors.affiliation: Divisions of Research Immunology/Bone Marrow Transplantation and Hematology/ Oncology, Department of Pediatrics, University of Southern California School of Medicine, Childrens Hospital Los Angeles, Los Angeles, CA, USA.
authors.forename: L
authors.initials: L
authors.lastname: Mascarenhas
authors.forename: R
authors.initials: R
authors.lastname: Stripecke
authors.forename: S S
authors.initials: SS
authors.lastname: Case
authors.forename: D
authors.initials: D
authors.lastname: Xu
authors.forename: K I
authors.initials: KI
authors.lastname: Weinberg
authors.forename: D B
authors.initials: DB
authors.lastname: Kohn
mesh_terms: D002411:Cation Exchange Resins / Q000494:pharmacology; D002469:Cell Separation; D018920:Coculture Techniques; D004274:DNA, Recombinant / Q000235:genetics; D004337:Drug Carriers / Q000494:pharmacology; D018274:Electroporation; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics; D049452:Green Fluorescent Proteins; D015497:HIV-1 / Q000235:genetics; D006801:Humans; D017989:Leukemia Virus, Gibbon Ape / Q000235:genetics; D008055:Lipids / Q000494:pharmacology; D008081:Liposomes; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D008979:Moloney murine leukemia virus / Q000235:genetics; D010714:Phosphatidylethanolamines / Q000494:pharmacology; D015452:Precursor B-Cell Lymphoblastic Leukemia-Lymphoma / Q000473:pathology* / Q000628:therapy; D011479:Protamines; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection; D014407:Tumor Cells, Cultured; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics
chemical_list: D002411:Cation Exchange Resins; D004274:DNA, Recombinant; D004337:Drug Carriers; D008055:Lipids; C086724:Lipofectamine; D008081:Liposomes; D008164:Luminescent Proteins; D010714:Phosphatidylethanolamines; D011479:Protamines; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; C020888:1,2-dielaidoylphosphatidylethanolamine