Gene delivery to human B-precursor acute lymphoblastic leukemia cells

Blood. 1998 Nov 15;92(10):3537-45.

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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cation Exchange Resins / pharmacology
  • Cell Separation
  • Coculture Techniques
  • DNA, Recombinant / genetics
  • Drug Carriers / pharmacology
  • Electroporation
  • Flow Cytometry
  • Gene Transfer Techniques*
  • Genes, Reporter
  • Genetic Therapy
  • Genetic Vectors / administration & dosage*
  • Genetic Vectors / genetics
  • Green Fluorescent Proteins
  • HIV-1 / genetics
  • Humans
  • Leukemia Virus, Gibbon Ape / genetics
  • Lipids / pharmacology
  • Liposomes
  • Luminescent Proteins / biosynthesis
  • Luminescent Proteins / genetics
  • Moloney murine leukemia virus / genetics
  • Phosphatidylethanolamines / pharmacology
  • Precursor B-Cell Lymphoblastic Leukemia-Lymphoma / pathology*
  • Precursor B-Cell Lymphoblastic Leukemia-Lymphoma / therapy
  • Protamines
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / genetics
  • Transfection
  • Tumor Cells, Cultured
  • Vesicular stomatitis Indiana virus / genetics

Substances

  • Cation Exchange Resins
  • DNA, Recombinant
  • Drug Carriers
  • Lipids
  • Lipofectamine
  • Liposomes
  • Luminescent Proteins
  • Phosphatidylethanolamines
  • Protamines
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins
  • 1,2-dielaidoylphosphatidylethanolamine