Heparin oligosaccharides bind L- and P-selectin and inhibit acute inflammation

Abstract

Initial attachment of leukocytes to the vessel wall at sites of inflammation is supported by a family of carbohydrate-binding adhesion molecules called the selectins. Selectin ligands include sialyl-Lewis x (sLex, Neu5Ac alpha 2–3Gal beta 1–4[Fuc alpha 1–3]GlcNAc--) and related structures. We report here that defined heparin oligosaccharides interact with the selectins. Heparin chains containing four or more monosaccharide residues inhibited the function of L- and P-selectin, but not E-selectin, in vitro. In a competition enzyme-linked immunosorbent assay measuring inhibition of solution-phase selectin-Ig fusion proteins (selectin-Ig) binding to immobilized bovine serum albumin-sLex neoglycoprotein, a heparin-derived tetrasaccharide mixture inhibited 50% of L- and P-selectin-Ig binding (IC50) at 200 +/- 40 mumol/L and 850 +/- 110 mumol/L, respectively. A single hexasulfated tetrasaccharide (delta UA2S alpha 1–4GlcNS6S alpha 1–4IdoA2S alpha 1- 4GlcNS6S) was particularly active against L- and P-selectin-Ig (IC50 = 46 +/- 5 mumol/L and 341 +/- 24 mumol/L). By comparison, the tetrasaccharide sLex was not inhibitory at concentrations up to 1 mmol/L. In cell adhesion assays, heparin tetrasaccharides reduced binding of neutrophils to COS cells expressing P-selectin but not to COS cells expressing E-selectin. They also blocked colon cancer cell adhesion to L- and P-selectin but not E-selectin. In a model of acute inflammation, intravenously administered heparin tetrasaccharides diminished influx of neutrophils into the peritoneal cavities of thioglycollate-treated mice. We conclude that heparin oligosaccharides, including non-anticoagulant tetrasaccharides, are effective L- and P- selectin inhibitors in vitro and have anti-inflammatory activity in vivo.

Intravascular granulocyte aggregates caused by the selectin‐binding carbohydrate fucoidin in pig kidneys

“…The importance of L-selectinmediated adhesion events in the ex vivo kidney reperfusion model may have been masked, in part, by L-selectin shedding secondary to leucocyte manipulation occurring during blood withdrawal and roller pumping. [39][40][41] Heparin, which has been shown to interfere with L-selectin ligand recognition, 42 showed only minor inhibition of L-selectin-dependent adhesion in vitro at concentrations used during ex vivo perfusion. However, although plasma heparin level analyses were below 20 U/mL, the possibility that the heparin used in our model could have modified the effect of fucoidin cannot be completely ruled out.…”

Regulation of interleukin‐8 binding and function by heparin and α2‐macroglobulin

“…These include strategic effects on several of the processes involved in inflammatory cell migration and accumulation in tissues. Inhibition of heparanase activity by low doses of heparin profoundly influenced lymphocyte trafficking [9] and heparin blocks the early adhesion-molecule dependent steps in the extravasation of leucocytes [35]. Further to the inhibitory effects of heparin on cytokine production [36,37], it is clear that heparin influences the activity of a number of cytokines and growth factors [38][39][40][41].…”

Oxidative stress induced by bloodless limb surgery on humans

“…In addition, heparin has been shown to attenuate the damage of the endothelial wall by neutrophils [35]. Several studies indicate that heparin inhibits the rolling, adherence and subsequent trafficking of leucocytes into tissues [36][37][38]. The drug has also been found to block the respiratory burst of neutrophils as well as the enzyme release from azurophilic granules [39,40].…”