Efficient adenovirus-mediated gene transduction of normal and leukemic hematopoietic cells.
We evaluated the efficiency of adenovirus-mediated gene transfer into normal and malignant human hematopoietic cells. An E-1 and E-3 deleted, replication-defective recombinant Ad.RSV beta gal vector was used and the transduction efficiency was studied at a multiplicity of infection of 13 p.f.u. per cell. Approximately 40-50% of normal monocytes were transduced, whereas purified normal resting T cells and B cells were resistant to infection. We showed that 50-80% of primary chronic myeloid leukemia cells (CML, n = 12) were efficiently transduced in contrast to CML, successful transduction of resting primary chronic B lymphocytic leukemia cells required appropriate preactivation of targeted cells. A novel protocol for the efficient transduction of adenovirus into B-CLL cells was presented. We showed that anti-CD40 mAb or CD40 ligand acts in synergy with rhIL-4 to enable the transduction of approximately 50-75% of B-CLL cells (B-CLL, n = 6). Expression of beta-galactosidase in transduced CML cells and B-CLL cells was detected for at least 15 days after transduction. The present studies underline the utility of adenovirus vectors for the construction of cytokine gene-modified tumor vaccines for the treatment of hematopoietic malignancies such as CML and B-CLL.
Huang
M R
MR
Department of Clinical Immunology, University Hospital, Uppsala, Sweden.
Olsson
M
M
Kallin
A
A
Pettersson
U
U
Tötterman
T H
TH
D000256:Adenoviridae / Q000235:genetics*; D019013:CD40 Antigens / Q000235:genetics; D018929:Cell Culture Techniques; D018014:Gene Transfer Techniques; D006801:Humans; D015847:Interleukin-4 / Q000235:genetics; D007763:Lac Operon; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000201:enzymology / Q000235:genetics* / Q000276:immunology; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000201:enzymology / Q000235:genetics*; D009000:Monocytes / Q000821:virology; D011994:Recombinant Proteins / Q000235:genetics; D014407:Tumor Cells, Cultured; D001616:beta-Galactosidase / Q000378:metabolism
D019013:CD40 Antigens; D011994:Recombinant Proteins; D015847:Interleukin-4; D001616:beta-Galactosidase