Syndecan-4 mediates antithrombin-induced chemotaxis of human peripheral blood lymphocytes and monocytes

Antithrombin inhibits chemokine-induced migration of neutrophils by activating heparan sulfate proteoglycan-dependent signaling. Whether antithrombin affects migration of other types of leukocytes is not known. We investigated the effects of antithrombin on spontaneous and chemokine-triggered migration of lymphocytes and monocytes from human peripheral blood in modified Boyden chamber micropore filter assays. Lymphocyte and monocyte populations from human peripheral blood were purified using magnetic antibody cell sorting. The signaling mechanisms required for antithrombin-dependent migration were studied using signaling enzyme blockers. Expression of heparan sulfate proteoglycan core protein was studied by RT-PCR and flow cytometry. The antithrombins used were Kybernin®P from human plasma and a monoclonal-antibody-purified preparation from this plasma. Pretreatment of lymphocytes and monocytes with antithrombin inhibited chemotaxis toward optimal concentrations of interleukin-8 or Rantes (regulated upon activation normal T-cell expressed and activated) at concentrations of antithrombin as low as 10 nU/ml. In the absence of the chemokines, direct exposure of cells to gradients of antithrombin stimulated migration. Effects of antithrombin were abolished by pretreating cells with heparinase-1, chondroitinase, sodium chlorate and anti-syndecan-4 antibodies. Expression of syndecan-4 mRNA and protein in monocytes and lymphocytes was demonstrated in RT-PCR and anti-syndecan-4 immunoreactivity assays, respectively. In the presence of pentasaccharide, antithrombin lost its effect on cells. Data indicate that antithrombin directly inhibits chemokine-stimulated migration of monocytes and lymphocytes via the effects of its heparin-binding site on cell surface syndecan-4 by activation of protein kinase C and Rho signaling.

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The core protein of the matrix-associated heparan sulfate proteoglycan binds to fibronectin.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(19)38948-3 ◽

1990 ◽

Vol 265 (15) ◽

pp. 8716-8724

Author(s):

A Heremans ◽

B De Cock ◽

J J Cassiman ◽

H Van den Berghe ◽

G David

Keyword(s):

Heparan Sulfate ◽

Core Protein ◽

Heparan Sulfate Proteoglycan ◽

Sulfate Proteoglycan ◽

The Core ◽

The Matrix

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Products of the unc-52 gene in Caenorhabditis elegans are homologous to the core protein of the mammalian basement membrane heparan sulfate proteoglycan.

Genes & Development ◽

10.1101/gad.7.8.1471 ◽

1993 ◽

Vol 7 (8) ◽

pp. 1471-1484 ◽

Cited By ~ 159

Author(s):

T M Rogalski ◽

B D Williams ◽

G P Mullen ◽

D G Moerman

Keyword(s):

Caenorhabditis Elegans ◽

Basement Membrane ◽

Heparan Sulfate ◽

Core Protein ◽

Heparan Sulfate Proteoglycan ◽

Sulfate Proteoglycan ◽

The Core

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Characterization of a novel heparan sulfate proteoglycan found in the extracellular matrix of liver sinusoids and basement membranes.

The Journal of Cell Biology ◽

10.1083/jcb.113.5.1231 ◽

1991 ◽

Vol 113 (5) ◽

pp. 1231-1241 ◽

Cited By ~ 33

Author(s):

C J Soroka ◽

M G Farquhar

Keyword(s):

Extracellular Matrix ◽

Basement Membrane ◽

Heparan Sulfate ◽

Rat Liver ◽

Core Protein ◽

Heparan Sulfate Proteoglycan ◽

Basement Membranes ◽

Sulfate Proteoglycan ◽

Sinusoidal Cell ◽

Space Of Disse

A novel heparan sulfate proteoglycan (HSPG) present in the extracellular matrix of rat liver has been partially characterized. Proteoglycans were purified from a high salt extract of total microsomes from rat liver and found to consist predominantly (approximately 90%) of HSPG. A polyclonal antiserum raised against this fraction specifically recognized HSPG by immunoprecipitation and immunoblotting. The intact, fully glycosylated HSPG migrated as a broad smear (150-300 kD) by SDS-PAGE, but after deglycosylation with trifluoromethanesulfonic acid only a single approximately 40-kD band was seen. By immunocytochemistry this HSPG was localized in the perisinusoidal space of Disse associated with irregular clumps of basement membrane-like extracellular matrix material, some of which was closely associated with the hepatocyte sinusoidal cell surface. It was also localized in biosynthetic compartments (rough ER and Golgi cisternae) of hepatocytes, suggesting that this HSPG is synthesized and deposited in the space of Disse by the hepatocyte. The anti-liver HSPG IgG also stained basement membranes of hepatic blood vessels and bile ducts as well as those of kidney and several other organs (heart, pancreas, and intestine). An antibody that recognizes the basement membrane HSPG found in the rat glomerular basement membrane did not precipitate the 150-300-kD rat liver HSPG. We conclude that the liver sinusoidal space of Disse contains a novel population of HSPG that differs in its overall size, its distribution and in the size of its core protein from other HSPG (i.e., membrane-intercalated HSPG) previously described in rat liver. It also differs in its core protein size from HSPG purified from other extracellular matrix sources. This population of HSPG appears to be a member of the basement membrane HSPG family.

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Matrix-associated heparan sulfate proteoglycan: core protein-specific monoclonal antibodies decorate the pericellular matrix of connective tissue cells and the stromal side of basement membranes.

The Journal of Cell Biology ◽

10.1083/jcb.109.6.3199 ◽

1989 ◽

Vol 109 (6) ◽

pp. 3199-3211 ◽

Cited By ~ 42

Author(s):

A Heremans ◽

B van der Schueren ◽

B de Cock ◽

M Paulsson ◽

J J Cassiman ◽

...

Keyword(s):

Extracellular Matrix ◽

Monoclonal Antibodies ◽

Epithelial Cell ◽

Heparan Sulfate ◽

Core Protein ◽

Structural Features ◽

Heparan Sulfate Proteoglycan ◽

Basement Membranes ◽

Lung Fibroblasts ◽

Sulfate Proteoglycan

Cultured human lung fibroblasts produce a large, nonhydrophobic heparan sulfate proteoglycan that accumulates in the extracellular matrix of the monolayer (Heremans, A., J. J. Cassiman, H. Van den Berghe, and G. David. 1988. J. Biol. Chem. 263: 4731-4739). A panel of four monoclonal antibodies, specific for four distinct epitopes on the 400-kD core protein of this extracellular matrix heparan sulfate proteoglycan, detects similar proteoglycans in human epithelial cell cultures. Immunohistochemistry of human tissues with the monoclonal antibodies reveals that these proteoglycans are concentrated at cell-matrix interfaces. Immunogold labeling of ultracryosections of human skin indicates that the proteoglycan epitopes are nonhomogeneously distributed over the width of the basement membrane. Immunochemical investigations and amino acid sequence analysis indicate that the proteoglycan from the fibroblast matrix shares several structural features with the large, low density heparan sulfate proteoglycan isolated from the Engelbreth-Holm-Swarm sarcoma. Thus, both epithelial cell sheets and individual mesenchymal cells accumulate a large heparan sulfate proteoglycan(s) at the interface with the interstitial matrix, where the proteoglycan may adopt a specific topological orientation with respect to this matrix.

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