In Vivo Ligand Specificity of E-selectin Binding to Multivalent Sialyl Lewisx N-linked Oligosaccharides

Abstract

The in vivo specificity for E-selectin binding to a panel of N-linked oligosaccharides containing a clustered array of one to four sialyl Lewis x (SLe x ; NeuAc␣2-3Gal[Fuc␣1-3]␤1-4GlcNAc) determinants was studied in mice. Following intraperitoneal dosing with lipopolysaccharide, radioiodinated tyrosinamide N-linked oligosaccharides were dosed i.v. and analyzed for their pharmacokinetics and biodistribution. Specific targeting was determined from the degree of SLe x oligosaccharide targeting relative to a sialyl oligosaccharide control. Oligosaccharides targeted the kidney with the greatest selectivity after a 4-h induction period following lipopolysaccharide dosing. Unique pharmacokinetic profiles were identified for SLe x biantennary and triantennary oligosaccharides but not for monovalent and tetraantennary SLe x oligosaccharides or sialyl oligosaccharide controls. Biodistribution studies established that both SLe x biantennary and triantennary oligosaccharides distributed to the kidney with 2-3-fold selectivity over sialyl oligosaccharide controls, whereas monovalent and tetraantennary SLe x oligosaccharides failed to mediate specific kidney targeting. Simultaneous dosing of SLe x biantennary or triantennary oligosaccharide with a mouse anti-E-selectin monoclonal antibody blocked kidney targeting, whereas co-administration with anti-P-selectin monoclonal antibody did not significantly block kidney targeting. The results suggest that SLe x biantennary and triantennary are N-linked oligosaccharide ligands for E-selectin and implicate E-selectin as a bivalent receptor in the murine kidney endothelium.

Serum Protein N-Glycosylation Changes with Rheumatoid Arthritis Disease Activity during and after Pregnancy

“…These structures may be extended by additional monosaccharides such as a bisecting N -acetylglucosamine, as well as galactoses, N -acetylneuraminic acids (sialic acids), and fucoses in a variety of different positions and linkages. This leads to a large N -glycan diversity, and may also lead to the formation of specific epitopes such as sialyl-Lewis X, which can be recognized by E-selectin ( 12 , 16 ).…”

E‐selectin targeting to visualize tumorsin vivo

“…The high affinity of liposomes to E-selectin might account for the difference in retention time of fluorescence between the SLX and anti-E-selectin antibody. Monovalent SLX to E-selectin is very weak (K d , mM) relative to the affinity of antibody to E-selectin (K d , nM), which results in efficient uptake into tumor tissue (23). Thus longer retention times of liposomes in tumors could be expected when liposomes are conjugated to anti-E-selectin antibody.…”

Design, synthesis, physical and chemical characterisation, and biological interactions of lectin-targeted latex nanoparticles bearing Gd–DTPA chelates: an exploration of magnetic resonance molecular imaging (MRMI)

“…On our nanoparticles, the targeting groups appear to be spaced almost regularly 15 nm apart. The density of N-glycans exposed at a cell surface may be 15,000/lm 2 (Chigaev et al 2001;D'Ambrosio et al 2002;Lecca et al 2004), or approaching 100,000/ lm 2 , taking the closest approach between two N-glycan antennae to be approximately 0.2 nm (Lee 1992;Thomas et al 1999). The approximately 4,000 targeting groups/ micrometre squared on the nanoparticle would find up to 100,000 ligands/micrometre squared on the cell surface.…”

“…Some of our SEM images may show nanoparticles rolling on cell surfaces, and this behaviour is to be expected when endothelium-targeted nanoparticles carried in the bloodstream make initial contact with the endothelium of the vessel wall. Leucocyte rolling on endothelial cells has been extensively researched, and its mechanics has been described quantitatively (Alon et al 1997;Thomas et al 1999). The comparison with nanoparticle rolling cannot be exact because of the size difference, the nanoparticles being approximately 100· linearly smaller than a leucocyte and the nanoparticles being passive whereas the leucocyte possesses an extremely active and responsive machinery.…”