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Laboratoire dImmunologie Cellulaire, Unité Mixte de Recherche 7627, Centre National de la Recherche Scientifique, Hôpital Pitié-Salpêtrière, Paris, France;
Unité Mixte de Recherche, Centre National de la Recherche Scientifique 144, Institut Curie, Paris, France; and
Laboratoire dImmunologie Cellulaire de lÉcole Pratique des Hautes Études, Hôpital Pitié-Salpêtrière, Paris, France
Whether thymic dendritic cells (DC) are phenotypically and
functionally distinct from the monocyte lineage DC is an important
question. Human thymic progenitors differentiate into T, NK, and DC.
The latter induce clonal deletion of autoreactive thymocytes and
therefore might be different from their monocyte-derived counterparts.
The cytokines needed for the differentiation of DC from thymic
progenitors were also questioned, particularly the need for GM-CSF. We
show that various cytokine combinations with or without GM-CSF
generated DC from CD34+CD1a- but not from
CD34+CD1a+ thymocytes. CD34+ thymic
cells generated far fewer DC than their counterparts from the cord
blood. The requirement for IL-7 was strict whereas GM-CSF was
dispensable but nonetheless improved the yield of DC. CD14+
monocytic intermediates were not detected in these cultures unless
macrophage-CSF (M-CSF) was added. Cultures in M-CSF generated
CD14-CD1a+ DC precursors but also
CD14+CD1a- cells. When sorted and recultured
in GM-CSF, CD14+ cells down-regulated CD14 and up-regulated
CD1a. TNF-
accelerated the differentiation of progenitors into DC
and augmented MHC class II transport to the membrane, resulting in
improved capacity to induce MLR. The trafficking of MHC class II
molecules was studied by metabolic labeling and immunoprecipitation.
MHC class II molecules were transported to the membrane in association
with invariant chain isoforms in CD14+ (monocyte)-derived
and in CD1a+ thymic-derived DC but not in monocytes. Thus,
thymic progenitors can differentiate into DC along a preferential
CD1a+ pathway but have conserved a CD14+
maturation capacity under M-CSF. Finally, CD1a+-derived
thymic DC and monocyte-derived DC share very close Ag-processing
machinery.
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