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Growth factors increase amphotropic retrovirus binding to human CD34+ bone
marrow progenitor cells
GM Crooks and DB Kohn
Division of Research Immunology and Bone Marrow Transplantation, Childrens
Hospital Los Angeles, CA 90027.
Gene transfer into human cells using murine amphotropic retroviral vectors
is the basic technique used in most current gene therapy studies. The
identity of the cell surface receptor for the amphotropic envelope remains
unknown and thus its importance in gene transfer is poorly understood. We
have measured specific retrovirus binding to cells to study amphotropic
virus receptor regulation in human CD34+ bone marrow (BM) progenitors and
primitive CD34+CD38- human hematopoietic cells. The rat monoclonal antibody
83A25 recognizes an epitope common to the envelope glycoprotein of all
classes of Moloney murine leukemia virus. Indirect fluorescent labeling of
83A25 allows flow cytometric analysis of specific virus-cell interactions
and is an indirect measure of specific receptors. Using this assay,
amphotropic virus binding to fresh CD34+ cells was minimal. However, when
CD34+ cells were cultured with or without growth factors for 4 days,
specific binding of amphotropic retrovirus was readily shown. Inclusion of
interleukin-3 (IL-3), IL-6, and Steel factor in cultures increased the
fluorescence associated with amphotropic virus binding by twofold to
four-fold (mean fold increase 2.7 +/- 0.84). Virus binding to CD34+CD38-
cells was shown only in those cells culture in IL-3, IL-6, and Steel
factor. These results suggest that certain cytokines may cause an increase
in the number and/or affinity of amphotropic receptors on primitive human
hematopoietic cells. Upregulation of viral receptor expression may be one
of the mechanisms by which cytokines enhance gene transfer into primitive
BM cells.
Volume 82,
Issue 11,
pp. 3290-3297,
12/01/1993
Copyright © 1993 by The American Society of Hematology
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