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Delineation of the dendritic cell lineage by generating large numbers of
Birbeck granule-positive Langerhans cells from human peripheral blood
progenitor cells in vitro
A Mackensen, B Herbst, G Kohler, G Wolff-Vorbeck, FM Rosenthal, H Veelken, P Kulmburg, HE Schaefer, R Mertelsmann and A Lindemann
Department of Medicine I (Hematology/Oncology), University Medical Center
Freiburg, Germany.
It is well established by in vivo and in vitro studies that dendritic cells
(DCs) originate from hematopoietic progenitor cells. However, the presumed
intermediate of Birbeck granule (BG)+ Langerhans cells (LCs) has not been
detected in cultures derived from bone marrow or peripheral blood
progenitor cells (PBPCs), thus contrasting with the data obtained with cord
blood. We show here that large numbers of BG+ LCs can be generated from
human CD34+ PBPCs in vitro, when granulocyte- macrophage colony-stimulating
factor and interleukin-4, potent promotors of LC/DC differentiation, are
combined with a cocktail of early acting hematopoietic growth factors. LCs
were found to emerge from CD33+CD11b+CD14- progenitor cells that they share
with the monocytic lineage. During culture, these cells exhibited a
sequence of dramatic morphologic changes, starting with a major increase in
granularity followed by an increase in size herein exceeding that of all
peripheral blood cells. At the same time, CD1a and major histocompatibility
complex class II expression were upregulated and virtually all CD1a++ cells
were BG+ by electron microscopy. With prolonged culture, CD1a was
downregulated on a major population of cells, paralleled by a loss of BG
and an increase of CD4, CD25, and CD80 expression that may correspond to
the maturation of epidermal LC in vitro. However, these cells were
consistently CD5- and did not exhibit changes in the CD45-isoform
expression during culture. The availability of large numbers of these
highly purified BG+ LCs and mature DCs allows for specific analysis of
these subpopulations and provides a source of potent antigen-presenting
cells from individual patients for vaccination protocols against infectious
or tumor- associated antigens.
Volume 86,
Issue 7,
pp. 2699-2707,
10/01/1995
Copyright © 1995 by The American Society of Hematology
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