Gene transfer into hematopoietic progenitor cells from normal and cyclic
hematopoietic dogs using retroviral vectors
MA Eglitis, PW Kantoff, JD Jolly, JB Jones, WF Anderson and CD Lothrop
Laboratory of Molecular Hematology, National Heart, Lung and Blood
Institute, Bethesda, MD.
The Moloney murine leukemia retrovirus-derived vector N2 was used to
transfer the bacterial NeoR gene (conferring resistance to the neomycin
analogue G418) into hematopoietic progenitor cells. Approximately 5% of day
seven CFU-GM were resistant to 2,000 micrograms/ml G418, using a
supernatant infection protocol in the absence of vector-producing cells. A
greater proportion of CFU-GM colonies were recovered relative to uninfected
controls as the stringency of selection was diminished. Enzyme activity was
detected in drug-resistant colonies, confirming that the resistant colonies
obtained after infection with N2 represented cells producing neomycin
phosphotransferase. Activity in the CFU-GM colonies approached 50% of that
of drug-resistant vector- producing cells on a per cell basis. To test the
hypothesis that more rapidly cycling bone marrow cells would be more
susceptible to vector infection, we treated progenitor cells obtained from
cyclic hematopoietic (CH) dogs with the N2 vector. Despite the increased
numbers of hematopoietic progenitor cells obtained from CH dogs, the
proportion of G418-resistant CFU-GM did not increase over that obtained
with N2-infected normal marrow. These results demonstrate that retroviral
vectors can be used to transfer and express exogenous genes in canine
hematopoietic progenitor cells.
Volume 71,
Issue 3,
pp. 717-722,
03/01/1988
Copyright © 1988 by The American Society of Hematology
