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Identification of upstream signals regulating interleukin-6 gene expression
during in vitro treatment of human B cells with pokeweed mitogen
D Chauhan, S Kharbanda, H Uchiyama, M Urashima, R Fragoso, J Sen, DW Kufe and KC Anderson
Division of Hematologic Malignancies, Dana-Farber Cancer Institute, Boston,
MA 02115.
The present studies have characterized the regulation of interleukin-6
(IL-6) gene expression during pokeweed mitogen (PWM)-driven human B- cell
differentiation. PWM induced an early and transient increase in the
expression of immediate-early response genes of the jun/fos leucine zipper
family (c-jun, jun B, c-fos, and fos-B). The induction of c-jun mRNA by PWM
was concentration dependent. Nuclear run-on assays showed that PWM
treatment is associated with an increased rate of c-jun gene transcription.
The induction of c-jun mRNA precedes the induction of IL- 6 gene expression
and IL-6 secretion by the B cells. c-Jun antisense, but not sense,
oligodeoxynucleotide (ODN) significantly decreases PWM- related B-cell (1)
proliferation; (2) IL-6 mRNA induction; (3) IL-6 secretion; and (4) nuclear
extract binding to AP-1 in electrophoretic mobility shift assay. In
contrast, c-Fos anti-sense ODN did not effect either IL-6 mRNA induction or
IL-6 secretion triggered in B cells by PWM. The results further show
activation of c-Raf-1 kinase in PWM- treated B cells. Raf-1 acts upstream
to mitogen-activated protein (MAP) kinase; therefore, studies were
performed to assay for MAP kinase activation in these cells. The results
show an increase in phosphorylation of myelin basic protein (MBP) and c-Jun
"Y" peptide in PWM-treated B cells. Taken together, these findings suggest
that PWM is able to initiate an intracytoplasmic signaling cascade in
normal human splenic B cells, which, at least in part, involves
serine/threonine protein kinases. These results show transient induction of
immediate- early response genes in B cells and support a potential role for
the c- jun gene product in regulation of IL-6 transcription and secretion.
Volume 84,
Issue 7,
pp. 2243-2252,
10/01/1994
Copyright © 1994 by The American Society of Hematology
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