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Blood, Vol. 92 No. 10 (November 15), 1998:
pp. 3537-3545
RAPID COMMUNICATION
Gene Delivery to Human B-Precursor Acute Lymphoblastic Leukemia Cells
Leo Mascarenhas,
Renata Stripecke,
Scott S. Case,
Dakun Xu,
Kenneth I. Weinberg, and
Donald B. Kohn
From the 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.
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.
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