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In vitro differentiation of CD34+ hematopoietic progenitor cells toward
distinct dendritic cell subsets of the birbeck granule and MIIC- positive
Langerhans cell and the interdigitating dendritic cell type
B Herbst, G Kohler, A Mackensen, H Veelken, P Kulmburg, FM Rosenthal, HE Schaefer, R Mertelsmann, P Fisch and A Lindemann
Department of Medicine I (Hematology/Oncology), University Medical Center
Freiburg, Germany.
We have demonstrated recently that Birbeck granule-positive Langerhans
cells (LC) can be derived from CD34+ peripheral blood progenitor cells in
the presence of a seven-cytokine cocktail (CC7-7). Here, we show that the
sequential use of early-acting hematopoietic growth factors, stem cell
factor, interleukin (IL)-3, and IL-6, followed on day 8 by differentiation
in the two-factor combination IL-4 plus granulocytemacrophage
colony-stimulating factor (GM-CSF) (CC4GM) is more efficient and allows the
cells to be arrested in the LC stage for more than 1 week while continuous
maturation occurs in CC7-7. Maturation of LC to interdigitating dendritic
cells (DC) could specifically be induced within 60 hours by addition of
tumor necrosis factor-alpha (20 ng/mL) or lipopolysaccharide (100 ng/mL).
Using LC that had been enriched to greater than 90% CD1a+ cells by an
immunoaffinity column, we were able to define clear-cut differences between
LC and DC that corroborate data of the respective cells derived from
epithelial borders (LC) or from lymph nodes (LN) and spleen (DC). Thus,
molecules and functions involved in antigen (AG) uptake and processing were
highly expressed in LC, while those involved in AG presentation were at
maximum in DC. LC were CD1a+2 DR+2, CD23+, CD36+, CD80-, CD86-, and CD25-,
while DC were CD1a+/- DR+3, CD23-, CD36-, CD80+, CD86+2, and CD25+, CD40
and CD32 were moderately expressed and nearly unchanged on maturation, in
contrast to monocyte-derived DC. Macropinocytosis of fluorescein
isothiocyanate-dextran was dominant in LC, as were multilamellar major
histocompatibility complex (MHC) class II compartments (MIICs), which were
detected by electron microscopy. The functional dichotomy of these cell
types was finally supported by testing the AG-presenting cell function for
tetanus toxoid to primed autologous T-cell lines, which was optimal when
cells were loaded with AG as LC and subsequently induced to become DC.
Volume 88,
Issue 7,
pp. 2541-2548,
10/01/1996
Copyright © 1996 by The American Society of Hematology
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