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 an d the transduction efficiency was studied at a multiplicity of infecti on of 13 pfu per cell. Approximately 40-50% of normal monocytes were t ransduced, whereas purified normal resting T cells and B cells were re sistant 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 lymphocy tic leukemia cells required appropriate preactivation of targeted cell s. 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 approximate ly 50-75% of B-CLL cells (B-CLL, n = 6). Expression of beta-galactosid ase 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 t umor vacines for the treatment of hematopoietic malignancies such as C ML and B-CLL.