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Ludwig Boltzmann Institute for Cell Biology and Immunobiology of the Skin, Department of Dermatology, University of Munster, Munster, Germany
Bone marrow-derived dendritic cells (BmDC) are potent APC and can
promote antitumor immunity in mice when pulsed with tumor Ag. This
study aimed to define the culture conditions and maturation stages of
BmDC that enable them to optimally function as APC in vivo. BmDC
cultured under various conditions (granulocyte-macrophage CSF (GM-CSF)
or GM-CSF plus IL-4 alone or in combination with Flt3 ligand, TNF-
,
LPS, or CD40 ligand (CD40L)) were analyzed morphologically,
phenotypically, and functionally and were tested for their ability to
promote prophylactic and/or therapeutic antitumor immunity. Each of the
culture conditions generated typical BmDC. Whereas cells cultured in
GM-CSF alone were functionally immature, cells incubated with CD40L or
LPS were mature BmDC, as evident by morphology, capacity to internalize
Ag, migration into regional lymph nodes, IL-12 secretion, and
alloantigen or peptide Ag presentation in vitro. The remaining cultures
exhibited intermediate dendritic cell maturation. The in vivo
Ag-presenting capacity of BmDC was compared with respect to induction
of both protective tumor immunity and immunotherapy of established
tumors, using the poorly immunogenic squamous cell carcinoma, KLN205.
In correspondence to their maturation stage, BmDC cultured in the
presence of CD40L exhibited the most potent immunostimulatory effects.
In general, although not entirely, the capacity of BmDC to induce an
antitumor immune response in vivo correlated to their degree of
maturation. The present data support the clinical use of mature, rather
than immature, tumor Ag-pulsed dendritic cells as cancer vaccines and
identifies CD40L as a potent stimulus to enhance their in vivo
Ag-presenting capacity.
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