autologous blood and marrow transplantation - Blog Science ...

624 Chapter 12: Gene Therapy
RESULTS
Transduction of primary AML cells
Leukocytes were isolated from peripheral blood of untreated AML patients and
infected using either a conventional adenovirus vector encoding the GFP or the
same vector complexed with a polyamidoamine dendramer compound (VPC, see
Methods). Infections were done in both the presence and absence of exogenous
cytokines, and cells were assayed for gene transfer efficiency 48-72 hours later.
Shown in Fig. 1 is an example of one infection comparing the two methods of gene
transfer. A shows the leukemic blast population (denoted by the box), and B, C, and
D show uninfected, adenovirus only, or VPC infection, respectively. Table 1
summarizes our initial findings with seven patient samples. In the presence of
exogenous growth factors, the VPC method yielded an average 79% gene transfer
efficiency (± 13%), and the adenovirus alone yielded an average of 50% gene
transfer (± 19%). In the absence of growth factors, the same gene transfer vehicles
yielded efficiencies of 71% (± 11%) and 44% (± 10%), respectively.
in
E —
32
o
in
FSC-Height
Uninfected
i 0%
ft
I M1
I
10 10 10
SFP
1000
10 10
Transduction of Primary
Leukemic Blast Cells
Figure 1. Flow cytometric analysis of leukemic cells transduced with adenovirus or VPC.
The upper left panel shows the forward vs. side angle light scatter of a leukemic leukocyte
population. The box designated R3 indicates the blast population. Cells within the R3 box are
shown for uninfected, adenovirus infected, or VPC infected samples (lower panels, left to
right). Flow cytometric analysis was performed 24 hours postinfection.