Cell-surface heparan sulfate proteoglycan-mediated regulation of human neutrophil migration by the serpin antithrombin III.
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.
Dunzendorfer
S
S
Department of Internal Medicine, Innsbruck University Hospital, Innsbruck, Austria, and Aventis Behring GmbH Research, Marburg, Germany.
Kaneider
N
N
Rabensteiner
A
A
Meierhofer
C
C
Reinisch
C
C
Römisch
J
J
Wiedermann
C J
CJ
D000911:Antibodies, Monoclonal / Q000276:immunology; D000990:Antithrombin III / Q000187:drug effects / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology*; D000991:Antithrombins / Q000494:pharmacology*; D001665:Binding Sites; D002478:Cells, Cultured / Q000187:drug effects; D002634:Chemotaxis, Leukocyte / Q000187:drug effects*; D019812:Heparan Sulfate Proteoglycans / Q000502:physiology*; D019763:Heparin Lyase / Q000494:pharmacology; D026541:Heymann Nephritis Antigenic Complex; D006629:Hirudins / Q000494:pharmacology; D006801:Humans; D007163:Immunosorbent Techniques; D016209:Interleukin-8 / Q000378:metabolism / Q000494:pharmacology; D017493:Leukocyte Common Antigens / Q000378:metabolism; D019272:Leukocyte Elastase / Q000494:pharmacology; D008562:Membrane Glycoproteins / Q000378:metabolism / Q000494:pharmacology; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000037:antagonists & inhibitors / Q000494:pharmacology; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects*; D009844:Oligosaccharides / Q000037:antagonists & inhibitors / Q000502:physiology; D010447:Peptide Hydrolases / Q000494:pharmacology; D011509:Proteoglycans / Q000378:metabolism; D023062:Receptors, Interleukin-8A / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism; D023063:Receptors, Interleukin-8B / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism; D000080506:Sialyl Lewis X Antigen; D015398:Signal Transduction; D013329:Structure-Activity Relationship; D053671:Syndecan-4
D000911:Antibodies, Monoclonal; D000991:Antithrombins; D019812:Heparan Sulfate Proteoglycans; D026541:Heymann Nephritis Antigenic Complex; D006629:Hirudins; D016209:Interleukin-8; D008562:Membrane Glycoproteins; D009844:Oligosaccharides; D011509:Proteoglycans; D023062:Receptors, Interleukin-8A; D023063:Receptors, Interleukin-8B; C508304:SDC4 protein, human; D000080506:Sialyl Lewis X Antigen; D053671:Syndecan-4; C046193:antithrombin III-protease complex; D009240:N-Formylmethionine Leucyl-Phenylalanine; D000990:Antithrombin III; D017493:Leukocyte Common Antigens; D010447:Peptide Hydrolases; D019272:Leukocyte Elastase; D019763:Heparin Lyase