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Departments of
*
Immunobiology,
Research Administration, and
Analytical Chemistry and Formulation, Immunex Corporation, Seattle, WA 98101; and
§
The Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia
Dendritic cells (DC) are potent APCs that can be characterized in
the murine spleen as CD11bhighCD11chigh or
CD11blowCD11chigh. Daily injection of mice of
Flt3 ligand (FL) into mice transiently expands both subsets of DC in
vivo, but the effect of administration of GM-CSF on the expansion of DC
in vivo is not well defined. To gain further insight into the role of
GM-CSF in DC development and function in vivo, we treated mice with
polyethylene glycol-modified GM-CSF (pGM-CSF) which has an increased
half-life in vivo. Administration of pGM-CSF to mice for 5 days led to
a 5- to 10-fold expansion of CD11bhighCD11chigh
but not CD11blowCD11chigh DC. DC from
pGM-CSF-treated mice captured and processed Ag more efficiently than DC
from FL-treated mice. Although both FL- and pGM-CSF-generated
CD11bhighCD11chigh DC were
CD8
-, a greater proportion of these DC from
pGM-CSF-treated mice were 33D1+ than from FL-treated mice.
CD11blowCD11chigh DC from FL-treated mice
expressed high levels of intracellular MHC class II. DC from both
pGM-CSF- and FL-treated mice expressed high levels of surface class II,
low levels of the costimulatory molecules CD40, CD80, and CD86 and were
equally efficient at stimulating allogeneic and Ag-specific T cell
proliferation in vitro. The data demonstrate that treatment with
pGM-CSF in vivo preferentially expands
CD11bhighCD11chigh DC that share phenotypic and
functional characteristics with FL-generated
CD11bhighCD11chigh DC but can be distinguished
from FL-generated DC on the basis of Ag capture and surface expression
of 33D1.
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