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Selection of retrovirally transduced hematopoietic cells using CD24 as a
marker of gene transfer
R Pawliuk, R Kay, P Lansdorp and RK Humphries
Terry Fox Laboratory, British Columbia Cancer Agency, Vancouver, Canada.
We have investigated the use of a cell surface antigen as a dominant
selectable marker to facilitate the detection and selection of retrovirally
infected target cells. The small coding region of the human cell surface
antigen CD24 (approximately 240 bp) was introduced into a
myeloproliferative sarcoma virus (MPSV)-based retroviral vector, which was
then used to infect day 4 5-fluorouracil (5-FU)-treated murine bone marrow
cells. Within 48 hours of termination of the infection procedure
CD24-expressing cells were selected by fluorescent- activated cell sorting
(FACS) with an antibody directed against the CD24 antigen. Functional
analysis of these cells showed that they included not only in vitro
clonogenic progenitors and day 12 colony- forming unit-spleen but also
cells capable of competitive long-term hematopoietic repopulation.
Double-antibody labeling studies performed on recipients of retrovirally
transduced marrow cells showed that some granulocytes, macrophages,
erythrocytes, and, to a lesser extent, B and T lymphocytes still expressed
the transduced CD24 gene at high levels 4 months later. No gross
abnormalities in hematopoiesis were detected in mice repopulated with
CD24-expressing cells. Our results show that the use of the CD24 cell
surface antigen as a retrovirally encoded marker enables the rapid,
efficient, and nontoxic selection in vitro of infected primary cells,
facilitates tracking and phenotyping of their progeny, and should provide a
unique tool to identify elements that regulate the expression of transduced
genes in the most primitive hematopoietic cells.
Volume 84,
Issue 9,
pp. 2868-2877,
11/01/1994
Copyright © 1994 by The American Society of Hematology
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