2.50
Hdl Handle:
http://hdl.handle.net/10541/86125
Title:
Heparin and heparan sulfate bind interleukin-10 and modulate its activity.
Authors:
Salek-Ardakani, Shahram; Arrand, John R; Shaw, David M; Mackett, Mike
Abstract:
Glycosaminoglycans (GAG) are a group of negatively charged molecules that have been shown to bind and directly regulate the bioactivity of growth factors and cytokines such as basic fibroblast growth factor, transforming growth factor-beta, IL-7, and interferon-gamma. The ability of GAG to interact with human IL-10 (hIL-10) and the effect of these interactions on its biologic activity were analyzed. It was demonstrated by affinity chromatography that hIL-10 binds strongly to heparin-agarose at physiological pH. Biosensor-based binding kinetic analysis indicated an equilibrium dissociation constant, K(d), of 54 nmol/L for this interaction. Human IL-10 stimulated CD16 and CD64 expression on the monocyte/macrophage population within peripheral blood mononuclear cells, with optimal concentrations between 1 and 10 ng/mL. Soluble heparin, heparan sulfate, chondroitin sulfate, and dermatan sulfate were shown to inhibit the hIL-10-induced expression of CD16 and CD64 in a concentration-dependent manner. Heparin and heparan sulfate were most effective with IC(50) values of 100 to 500 microg/mL. Considerably higher concentrations of dermatan sulfate and chondroitin 4-sulfate were required with an IC(50) of 2,000 to 5,000 microg/mL, whereas chondroitin 6-sulfate was essentially inactive. The antagonistic effect of heparin on hIL-10 activity was shown to be dependent on N-sulfation, inasmuch as de-N-sulfated heparin had little or no inhibitory effect on the IL-10- induced expression of CD16, whereas the effect of de-O-sulfated heparin was comparable to that of unmodified heparin. Furthermore, the inhibition of cell-bound proteoglycan sulfation reduced the hIL-10-mediated expression of CD16 molecules on monocytes/macrophages. Taken together, these findings support the hypothesis that soluble and cell-surface GAG and, in particular, their sulfate groups are important in binding and modulation of hIL-10 activity. (Blood. 2000;96:1879-1888)
Affiliation:
CRC Molecular Biology Group, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Withington, Manchester, UK. ssalek@picr.man.ac.uk
Citation:
Heparin and heparan sulfate bind interleukin-10 and modulate its activity. 2000, 96 (5):1879-88 Blood
Journal:
Blood
Issue Date:
1-Sep-2000
URI:
http://hdl.handle.net/10541/86125
PubMed ID:
10961890
Type:
Article
Language:
en
ISSN:
0006-4971
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorSalek-Ardakani, Shahramen
dc.contributor.authorArrand, John Ren
dc.contributor.authorShaw, David Men
dc.contributor.authorMackett, Mikeen
dc.date.accessioned2009-11-13T13:15:19Z-
dc.date.available2009-11-13T13:15:19Z-
dc.date.issued2000-09-01-
dc.identifier.citationHeparin and heparan sulfate bind interleukin-10 and modulate its activity. 2000, 96 (5):1879-88 Blooden
dc.identifier.issn0006-4971-
dc.identifier.pmid10961890-
dc.identifier.urihttp://hdl.handle.net/10541/86125-
dc.description.abstractGlycosaminoglycans (GAG) are a group of negatively charged molecules that have been shown to bind and directly regulate the bioactivity of growth factors and cytokines such as basic fibroblast growth factor, transforming growth factor-beta, IL-7, and interferon-gamma. The ability of GAG to interact with human IL-10 (hIL-10) and the effect of these interactions on its biologic activity were analyzed. It was demonstrated by affinity chromatography that hIL-10 binds strongly to heparin-agarose at physiological pH. Biosensor-based binding kinetic analysis indicated an equilibrium dissociation constant, K(d), of 54 nmol/L for this interaction. Human IL-10 stimulated CD16 and CD64 expression on the monocyte/macrophage population within peripheral blood mononuclear cells, with optimal concentrations between 1 and 10 ng/mL. Soluble heparin, heparan sulfate, chondroitin sulfate, and dermatan sulfate were shown to inhibit the hIL-10-induced expression of CD16 and CD64 in a concentration-dependent manner. Heparin and heparan sulfate were most effective with IC(50) values of 100 to 500 microg/mL. Considerably higher concentrations of dermatan sulfate and chondroitin 4-sulfate were required with an IC(50) of 2,000 to 5,000 microg/mL, whereas chondroitin 6-sulfate was essentially inactive. The antagonistic effect of heparin on hIL-10 activity was shown to be dependent on N-sulfation, inasmuch as de-N-sulfated heparin had little or no inhibitory effect on the IL-10- induced expression of CD16, whereas the effect of de-O-sulfated heparin was comparable to that of unmodified heparin. Furthermore, the inhibition of cell-bound proteoglycan sulfation reduced the hIL-10-mediated expression of CD16 molecules on monocytes/macrophages. Taken together, these findings support the hypothesis that soluble and cell-surface GAG and, in particular, their sulfate groups are important in binding and modulation of hIL-10 activity. (Blood. 2000;96:1879-1888)en
dc.language.isoenen
dc.subject.meshAmino Acid Sequence-
dc.subject.meshAntigens, CD-
dc.subject.meshBinding, Competitive-
dc.subject.meshCells, Cultured-
dc.subject.meshChlorates-
dc.subject.meshChromatography, Affinity-
dc.subject.meshDose-Response Relationship, Drug-
dc.subject.meshFlow Cytometry-
dc.subject.meshGlycosaminoglycans-
dc.subject.meshHeparin-
dc.subject.meshHeparitin Sulfate-
dc.subject.meshHumans-
dc.subject.meshInterleukin-10-
dc.subject.meshKinetics-
dc.subject.meshLeukocytes, Mononuclear-
dc.subject.meshMacrophages-
dc.subject.meshMolecular Sequence Data-
dc.subject.meshMonocytes-
dc.subject.meshReceptors, IgG-
dc.subject.meshRecombinant Proteins-
dc.subject.meshSepharose-
dc.titleHeparin and heparan sulfate bind interleukin-10 and modulate its activity.en
dc.typeArticleen
dc.contributor.departmentCRC Molecular Biology Group, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Withington, Manchester, UK. ssalek@picr.man.ac.uken
dc.identifier.journalBlooden

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