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Secretion of cytokines (interleukins-1 alpha, -3, and -6 and
granulocyte-macrophage colony-stimulating factor) by normal human bone
marrow megakaryocytes
C Wickenhauser, J Lorenzen, J Thiele, A Hillienhof, K Jungheim, B Schmitz, ML Hansmann and R Fischer
Institute of Pathology, University of Cologne, Germany.
The effects of cytokine stimulation [recombinant human interleukin (rhIL)-1
alpha, rhIL-3, rhIL-6, rhIL-11, and rh granulocyte-macrophage
colony-stimulating factor (GM-CSF)] on the secretory activity of normal
human megakaryocytes were studied by means of the reverse hemolytic plaque
assay (RHPA) in enriched cell preparations. This test facilitates an
extremely sensitive determination of cytokine secretion at the single-cell
level, together with the clear-cut identification of each immunostained
(CD61) secretory active megakaryocyte. Moreover, the reverse
transcriptase-polymerase chain reaction (RT-PCR) was used to investigate
the expression of IL-6, IL-6 receptor (IL-6R), IL-9, IL-10, IL-12, and
IL-13 mRNA in highly concentrated megakaryocyte preparations. In comparison
with the spontaneous secretion rate, stimulation with rhIL-1 alpha, rhIL-6,
and rhGM-CSF failed to induce a significant increase in the release of
cytokines by CD61+ cells. On the other hand, both rhIL-3 and, in a less
pronounced way, rhIL-11 exerted a marked effect on IL-6 secretion.
Additionally, after stimulation with rhIL-3, a significant enhancement of
the secretion of IL-3 and GM-CSF, but not of IL-1 alpha, could be observed.
Using the RT-PCR, a significant induction of IL-6 expression could be
appreciated in the enriched megakaryocyte population (60% to 80%)
stimulated with rhIL-3. The results of this study provide persuasive
evidence that a number of cytokines are synthesized and secreted by human
megakaryocytes and not only by hematopoietic stroma cells. These data
suggest the existence of autocrine and paracrine mechanisms that may
influence maturation and differentiation of megakaryocytes as well as act
on various stroma cells to sustain an appropriate hematopoietic
micro-environment.
Volume 85,
Issue 3,
pp. 685-691,
02/01/1995
Copyright © 1995 by The American Society of Hematology
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