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Transduction of Primitive Hematopoietic Cells
With Adenovirus-Polycation Gene Transfer Vehicles:
New Strategies for Management
of Hematologic Disease
Craig T. Jordan, Dianna S. Howard, Shi-Fu Zhao,
John R. Yannelli, Barry Grimes
Markey Cancer Center, University of Kentucky Medical Center, Lexington, KY
ABSTRACT
Gene transfer into primitive hematopoietic cells has been proposed as a means to
achieve a variety of therapeutic results. Unfortunately, many types of hematopoietic
cells have proven to be largely refractory to standard methods of gene transfer.
Consequently, we have investigated the use of new strategies to mediate highefficiency
transduction of human hematopoietic tissue. Previously, we have shown
that under the appropriate conditions, adenoviral vectors can achieve limited gene
transfer into primitive CD34 +
cells (Neering SJ, Hardy SF, Minamoto D, Spratt SK,
Jordan CT: Transduction of primitive human hematopoietic cells with recombinant
adenovirus vectors. Blood 88:1147-1155, 1996). To extend this approach, we have
examined the use of polycationic compounds as a means to enhance the natural gene
delivery capacity of adenoviral vectors. These studies have shown that a vehicle
composed of adenoviral particles, coated with a polyamidoamine dendramer, is a
highly efficient complex for the transduction of primitive human hematopoietic
cells. To develop this method of gene transfer to be useful for immunotherapy
strategies, we tested the virus-polycation complex (VPC) for transduction of
primary myeloid leukemia cells. Using a VPC encoding a green fluorescent protein
(GFP) reporter gene, our data show that 79% (± 13, n=7) of primary leukemic blast
cells can be transduced using the VPC method. Moreover, approximately 70-80%
of the gene transfer activity occurs within the first 2 hours of the infection period,
and gene expression can be detected in as little as 3 hours. Also, cells transduced by
this method continue to express the transgene for at least 6-7 days. Therefore, we
suggest that this technique will be a highly effective strategy for the introductory of
immune regulatory genes (such as CD80 and granulocyte-macrophage colonystimulating
factor [GM-CSF]) into primary tumor cells for the purpose of generating
antileukemia vaccines.
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