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Generation of human dendritic cells/Langerhans cells from circulating CD34+
hematopoietic progenitor cells
D Strunk, K Rappersberger, C Egger, H Strobl, E Kromer, A Elbe, D Maurer and G Stingl
Division of Immunology, Allergy and Infectious Diseases, Vienna
International Research Cooperation Center (VIRCC), University of Vienna
Medical School, Austria.
Human Langerhans cells (LC) are CD1a+ dendritic cells (DC) that function as
potent antigen-presenting cells for primary and secondary immune responses.
Limitations in DC/LC numbers, imposed by difficult and tedious isolation
procedures, have so far precluded their use as immunogens in the generation
and/or augmentation of host responses against various pathogens. Therefore,
we have developed a procedure for the generation of human DC/LC from CD34+
hematopoietic progenitor cells (HPC) isolated (mean: 0.7 x 10(6)/ buffy
coat and 2.6 x 10(6)/leukapheresis product) and purified ( > 95%) from
the peripheral blood of healthy adults. In vitro stimulation of these cells
with granulocyte-macrophage colony-stimulating factor (GM-CSF) and tumor
necrosis factor (TNF)-alpha led to their vigorous proliferation and
differentiation resulting in the emergence of CD45+/CD68+/CD3-/CD19- /CD56-
leukocytes some of which (mean: 12%) express CD1a and exhibit anti-CD4 and
anti-major histocompatibility complex (MHC) class II reactivity. These
CD1a- leukocytes include (1) LC as evidenced by the presence of Birbeck
granules (BG), (2) CD14+ monocytes, and (3) Birbeck granule-negative cells
with a dendritic morphology. Addition of interleukin (IL)-4 to the cytokine
cocktail interfered with the development of monocytes and led to a
reduction in the overall yield but, on the other hand, resulted in an
increased percentage of CD1a+ cells (mean: 24%) among all cells generated.
In vitro generated CD1a+, but not CD1a- HPC-derived cells are potent
stimulators of the primary mixed leukocyte reaction and, as such, promising
candidates for vaccination purposes.
Volume 87,
Issue 4,
pp. 1292-1302,
02/15/1996
Copyright © 1996 by The American Society of Hematology
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