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Transduction of primitive human hematopoietic cells with recombinant
adenovirus vectors
SJ Neering, SF Hardy, D Minamoto, SK Spratt and CT Jordan
Somatix Therapy Corp, Alameda, CA 94501, USA.
We have examined the ability of recombinant adenoviral vectors to transduce
human hematopoietic cells. Our findings indicate that adenovirus readily
infects a large proportion of CD34+ cells. Using adenovirus vectors that
transduce either a lacZ or an alkaline phosphatase reporter gene, we
observed up to 45% of total CD34+ cells infected. Upon more detailed
analysis, we observed comparable levels of transduction for CD34+/CD38-
cells and for CD34+ cells in G(zero) phase of the cell cycle. Importantly,
exposure to adenovirus resulted in negligible levels of toxicity as assayed
by propidium iodide staining and colony-forming ability. Using adenovirus
vectors, we also describe a model system for regulated gene expression in
early hematopoietic tissues. CD34+ cells were simultaneously infected with
two viruses, one carrying a TetR/VP16 transactivator (tTA) and the second
carrying a tTA- dependent lacZ reporter gene. Using this approach, beta-gal
expression was only observed upon coinfection with the transactivator
vector. In addition, as shown previously (Gossen and Bujard, Proc Natl Acad
Sci USA 89:5547, 1992), tetracycline was able to inhibit tTA mediated
induction, thereby providing an effective means to regulate expression of
the reporter gene. We conclude that recombinant adenovirus is an effective
vehicle for transiently expressing genes in primitive human hematopoietic
cells.
Volume 88,
Issue 4,
pp. 1147-1155,
08/15/1996
Copyright © 1996 by The American Society of Hematology
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