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Interleukin-6 and its receptor are expressed by human megakaryocytes: in
vitro effects on proliferation and endoreplication
S Navarro, N Debili, JP Le Couedic, B Klein, J Breton-Gorius, J Doly and W Vainchenker
INSERM U91, Hopital Henri Mondor, Creteil, France.
Interleukin-6 (IL-6) is a pleiotropic cytokine that plays an important role
in the megakaryocytic differentiation. Recently, we have observed that IL-6
is synthesized by several human cell lines with megakaryocytic features. In
this study, we have investigated whether a similar phenomenon occurs during
normal megakaryocytic differentiation. Human megakaryocytes (MK) were
obtained by culturing normal marrow in liquid culture with aplastic plasma
(AP). First, an IL-6 secretion in bone marrow culture enriched in MK as
well as in purified MK populations was demonstrated by a biologic assay.
Second, IL-6 mRNA was detected in a purified population of MK by the
polymerase chain reaction and dot blot analysis. IL-6 mRNA and protein were
undetectable in platelets. Third, in situ hybridization procedure
demonstrated the presence of IL-6 mRNA in individual immature MK. Fourth,
IL-6 protein was detected in MK at the unicellular level by an
immunoalkaline phosphatase technique using a monoclonal antibody against
IL-6. Furthermore, the presence of IL-6 receptor (IL-6-R) on MK was
demonstrated by in situ hybridization using an IL-6-R probe and in situ
autoradiography after binding with [125I]-labeled recombinant IL-6. The
IL-6 endogenously produced in liquid cultures containing normal human
plasma or AP was subsequently neutralized. This resulted in a 50% decrease
of the MK growth with a minor shift in the ploidy distribution toward lower
values. In semisolid cultures the addition of anti-IL-6 antibodies led to a
42% decrease in colony number in cultures stimulated by IL-3 but not in
other conditions of culture. These results suggest that normal human
megakaryocytopoiesis might be regulated in part by an IL-6 autocrine loop.
Volume 77,
Issue 3,
pp. 461-471,
02/01/1991
Copyright © 1991 by The American Society of Hematology
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