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Heparin and heparan sulfate bind interleukin-10 and modulate its activity
S Salek-Ardakani1, J R Arrand, D Shaw
1CRC Molecular Biology Group, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Withington, Manchester, UK. ssalek@picr.man.ac.uk
Blood
|August 29, 2000
Summary
Glycosaminoglycans (GAGs) bind human interleukin-10 (hIL-10), modulating its activity. Sulfation of GAGs, particularly N-sulfation, is crucial for this interaction and the inhibition of hIL-10-induced immune cell marker expression.
Area of Science:
- Immunology
- Biochemistry
- Molecular Biology
Background:
- Glycosaminoglycans (GAGs) are known to regulate growth factors and cytokines.
- The interaction between GAGs and human interleukin-10 (hIL-10) and its functional consequences were not well understood.
Purpose of the Study:
- To investigate the binding of GAGs to hIL-10.
- To determine the effect of GAGs on the biological activity of hIL-10.
- To elucidate the structural requirements of GAGs for hIL-10 interaction.
Main Methods:
- Affinity chromatography to assess hIL-10 binding to heparin-agarose.
- Biosensor-based kinetic analysis to determine binding constants.
- Flow cytometry to measure CD16 and CD64 expression on monocytes/macrophages.
- Inhibition assays using various soluble GAGs and modified heparins.
Main Results:
- hIL-10 strongly binds to heparin-agarose with a K(d) of 54 nmol/L.
- hIL-10 stimulates CD16 and CD64 expression on monocytes/macrophages.
- Soluble heparin, heparan sulfate, dermatan sulfate, and chondroitin sulfate inhibit hIL-10-induced marker expression in a dose-dependent manner.
- Heparin and heparan sulfate were the most potent inhibitors, with N-sulfation being critical for heparin's inhibitory activity.
Conclusions:
- Soluble and cell-surface GAGs bind and modulate hIL-10 activity.
- The sulfate groups, particularly N-sulfates, on GAGs are essential for binding and inhibiting hIL-10.
- GAGs play a significant role in regulating the immune response mediated by hIL-10.