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1
*
Department of Hematology and Oncology, Clinical Sciences for Pathological Organs, Graduate School of Medicine, and
College of Medical Technology, Kyoto University, Sakyo-ku, Kyoto;
National Institute of Infectious Diseases, Tokyo; and
§
Department of Pharmacology, Cancer Research Institute, Kanazawa University, Kanazawa, Japan
TNF-
is implicated in the initiation of cytokine cascades in
various inflammatory settings. To assess the interactions of multiple
cytokines at the level of inflammatory effector cells, we examined the
effects of TNF- on the expression of two IL-8Rs (CXCR1 and CXCR2) on
polymorphonuclear leukocytes (PMNs). TNF- decreased the surface
expression of CXCR2 in a dose- and time-dependent manner. In contrast,
CXCR1 expression was not affected by TNF-. The release of CXCR2 into
the supernatant of TNF--treated PMNs was detected by immunoblotting
and immuno-slot-blot analyses, suggesting that the down-regulation of
CXCR2 was caused mainly by shedding from the cell surface. The CXCR2
down-regulation was inhibited by PMSF and aprotinin, supporting the
hypothesis that the shedding was mediated by serine protease(s). The
intracellular Ca2+ mobilization and chemotaxis in
response to IL-8 were suppressed by the pretreatment of PMNs with
TNF-, indicating that the decrease in CXCR2 was reflected in the
decreased functional responses to IL-8. In contrast, the
O2- release, which is mediated by CXCR1, was not
suppressed by TNF-. The treatment of whole blood with TNF- also
caused a significant reduction in CXCR2 and markedly suppressed
intracellular Ca2+ mobilization and chemotaxis in response
to IL-8, while enhancing the O2- release. These
findings suggest that TNF- down-regulates CXCR2 expression on PMNs
and modulates IL-8-induced biologic responses, leading to the
intravascular retention of PMNs with an enhanced production of reactive
oxygen metabolites.
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