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Blood, 15 February 2001, Vol. 97, No. 4, pp. 1079-1085
PHAGOCYTES
Cell-surface heparan sulfate proteoglycan-mediated regulation of
human neutrophil migration by the serpin antithrombin III
Stefan Dunzendorfer,
Nicole Kaneider,
Andrea Rabensteiner,
Christian Meierhofer,
Christina Reinisch,
Jürgen Römisch, and
Christian J. Wiedermann
From the Department of Internal Medicine, Innsbruck
University Hospital, Innsbruck, Austria, and Aventis Behring GmbH
Research, Marburg, Germany.
The serpin antithrombin III (AT III) is reported to have
hemostasis-regulating and anti-inflammatory properties. To determine its ability to influence thrombin-independent leukocyte responses, the
direct effects of the AT III concentrate Kybernin P and a monoclonal
antibody-purified AT III on neutrophil migration were studied.
Chemotactic activity of human neutrophils isolated from the blood of
healthy donors was determined in modified Boyden microchemotaxis
chambers, and binding studies were performed according to standard
experimental protocols. Preincubation in vitro of neutrophils with
Kybernin P or immune-adsorbed AT III significantly deactivated
migration toward fMet-Leu-Phe, or interleukin-8 (IL-8), in a
concentration-dependent manner. In the absence of additional attractants, neutrophils exhibited a migratory response toward gradients of AT III preparations. True chemotaxis was confirmed in
checkerboard assays. Analyses revealed that the AT III heparin-binding site interacts with neutrophil membrane-associated heparan sulfate proteoglycan receptors. Mechanisms of intracellular signaling differed;
the deactivation of IL-8-induced chemotaxis resulted from
tyrphostin-sensitive interactions of AT III-signaling with the IL-8
signal transduction pathway, whereas AT III-induced chemotaxis involved protein kinase C and phosphodiesterases. Signaling
similarities between AT III and the proteoglycan syndecan-4 may suggest
the binding of AT III to this novel type of membrane receptor. Under physiological conditions, AT III may prevent neutrophils from premature
activation. Moreover, the systemic administration of AT III
concentrate could have beneficial effects in combating systemic inflammation.
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