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Human macrophage colony-stimulating factor is expressed at and shed from
the cell surface
DP Tuck, DP Cerretti, A Hand, A Guha, S Sorba and N Dainiak
Department of Medicine, Royal Victoria Hospital, McGill University,
Montreal, Canada.
Surface membrane-associated growth factors are being recognized as
important for developmental processes, including cell assembly,
differentiation, and growth. To investigate the role of membrane-bound
macrophage colony-stimulating factor (M-CSF) in myelopoiesis, and whether
this factor is released from the cell surface in association with shed
membrane-derived vesicles, COS-1 cells were transfected with cDNAs for
M-CSF-tau (containing the transmembrane domain) or a soluble mutant form of
the molecule lacking the transmembrane domain ([s]M-CSF- alpha). COS-1
cells transfected with either cDNA released activity into the spent culture
medium. Conditioned medium was separated by centrifugation into
supernatants and pellets were found to contain plasma membrane-derived
vesicles by transmission electron microscopy. When medium fractions were
assayed in marrow cultures, activity was localized to shed plasma
membrane-derived vesicles in medium conditioned by cells transfected with
cDNA for M-CSF-tau and in the vesicle-free supernatants of medium
conditioned by cells transfected with cDNA for [s]M-CSF-alpha. In addition,
nuclear, mitochondrial, and plasma membrane subfractions of stably
transfected cells were prepared and assayed for activity.
Concentration-dependent stimulation of macrophage colony formation was
observed with purified plasma membranes (but not nuclear or cytosolic
proteins) from cells transfected with cDNA for M-CSF-tau. By contrast,
membranes from untransfected cells and cells transfected with cDNA for
[s]M-CSF-alpha or control DNA expressed no activity. Together, the data
indicate that human M-CSF is expressed at the cell surface and exfoliated
in association with surface membrane- derived vesicles.
Volume 84,
Issue 7,
pp. 2182-2188,
10/01/1994
Copyright © 1994 by The American Society of Hematology
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