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Blood, Vol. 93 No. 1 (January 1), 1999:
pp. 341-349
Cytokine-Specific Activation of Distinct Mitogen-Activated
Protein Kinase Subtype Cascades in Human Neutrophils
Stimulated by Granulocyte Colony-Stimulating Factor,
Granulocyte-Macrophage Colony-Stimulating Factor, and Tumor Necrosis
Factor-
Kenichi Suzuki,
Masayuki Hino,
Fumihiko Hato,
Noriyuki Tatsumi, and
Seiichi Kitagawa
From the Departments of Physiology and Clinical Hematology, Osaka
City University Medical School, Osaka, Japan.
To clarify the differences of the signaling pathways used by
granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), and tumor necrosis factor- (TNF), we investigated activation of mitogen-activated protein kinase
(MAPK) subtype cascades in human neutrophils stimulated by these
cytokines. G-CSF exclusively tyrosine-phosphorylated extracellular signal-regulated kinase (ERK). GM-CSF
tyrosine-phosphorylated ERK strongly and p38 MAPK weakly, whereas TNF
tyrosine-phosphorylated p38 MAPK strongly and ERK weakly. Consistent
with these findings, MEK, an upstream kinase of ERK, was phosphorylated
by G-CSF, GM-CSF, and TNF, whereas MKK3/MKK6, an upstream kinase of p38
MAPK, was phosphorylated by GM-CSF and TNF, but not by G-CSF. The
potency of these cytokines to phosphorylate ERK and MEK was GM-CSF > G-CSF > TNF, whereas that to phosphorylate p38 MAPK and MKK3/MKK6 was TNF > GM-CSF. C-Jun amino-terminal kinase (JNK) was not
tyrosine-phosphorylated by any cytokine despite the existence of JNK
proteins in human neutrophils, whereas it was tyrosine-phosphorylated
by TNF in undifferentiated and all-trans retinoic acid-differentiated
HL-60 cells. Increased phosphorylation of ERK or p38 MAPK was detected within 1 to 5 minutes after stimulation with each cytokine and was
dependent on the concentrations of cytokines used. MEK inhibitor (PD98059) reduced tyrosine phosphorylation of ERK, but not p38 MAPK,
induced by G-CSF, GM-CSF, or TNF. GM-CSF- or TNF-induced superoxide
(O2 ) release was inhibited by p38 MAPK
inhibitor (SB203580) in a dose-dependent manner, suggesting the
possible involvement of p38 MAPK in GM-CSF- or TNF-induced
O2 release. The results indicate that G-CSF,
GM-CSF, and TNF activate the overlapping but distinct MAPK subtype
cascades in human neutrophils and suggest that the differential
activation of ERK and p38 MAPK cascades may explain the differences of
the effects of these cytokines on human neutrophil functions.
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