Inhibition of monocyte, lymphocyte, and neutrophil adhesion to endothelial cells by human milk oligosaccharides

Summary

Excessive leukocyte infiltration causes severe tissue damage in a variety of inflammatory diseases. The initial step in leukocyte extravasation is mediated by selectins and oligosaccharides on their glycoconjugate ligands. Human milk is a rich source of lactose-derived oligosaccharides that are partly absorbed in the intestine and excreted with the urine. As these components contain binding determinants for the selectins we investigated whether human milk oligosaccharides are able to affect leukocyte rolling and adhesion to endothelial cells under dynamic conditions. Therefore, monocytes, lymphocytes, or neutrophils isolated from human peripheral blood were passed over TNF-α-activated HUVEC under shear stress. The influence of different oligosaccharide fractions was determined by video-micros-copy and compared with the effects of various individual oligosaccharides. Within a physiological range (12.5-125 µg/ml) the acidic fraction significantly inhibited leukocyte rolling and adhesion (up to 24.0% and 52.8%, respectively) in a concentrationdependent manner. These effects were even more pronounced than those achieved by soluble sialyl-Lewis x, a physiological binding determinant for selectins. Several active components within the oligosaccharide fraction of human milk were identified, e.g. 3’-sialyl-lactose and 3’-sialyl-3-fucosyl-lactose. These results indicate that specific oligosaccharides in human milk may serve as anti-inflammatory components and might therefore contribute to the lower incidence of inflammatory diseases in human milk-fed infants.

Keywords

Selectins - leukocytes - oligosaccharides - human milk - inflammation

• References

• 1 Springer TA. Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell 1994; 76: 301-14.
• 2 Varki A. Selectin ligands. Proc Natl Acad Sci 1994; 91: 7390-7.
• 3 Ley K. Integration of inflammatory signals by rolling neutrophils. Imm Rev 2002; 186: 8-18.
• 4 Rhee JS, Santoso S, Herrmann M. et al. New aspects of integrin-mediated leukocyte adhesion in inflammation: regulation by haemostatic factors and bacterial products. Curr Mol Med 2003; 03: 387-92.
• 5 Carden DL, Granger DN. Pathophysiology of ischaemia-reperfusion injury. J Pathol 2000; 190: 255-66.
• 6 Schon MP, Krahn T, Schon M. et al. Efomycine M, a new specific inhibitor of selectin, impairs leukocyte adhesion and alleviates cutaneous inflammation. Nat Med 2002; 08: 366-72.
• 7 Laroux FS, Grisham MB. Immunological basis of inflammatory bowel disease: role of the microcirculation. Microcirculation 2001; 08: 283-301.
• 8 Weyrich AS, Ma XY, Lefer DJ. et al. In vivo neutralization of P-selectin protects feline heart and endothelium in myocardial ischemia and reperfusion injury. J Clin Invest 1993; 91: 2620-9.
• 9 Ma XL, Weyrich AS, Lefer DJ. et al. Monoclonal antibody to L-selectin attenuates neutrophil accumulation and protects ischemic reperfused cat myocardium. Circulation 1993; 88: 649-58.
• 10 Simanek EE, McGarvey GJ, Jablonowski JA. et al. Selectin-carbohydrate interactions: From natural ligands to designed mimics. Chem Rev 1998; 98: 833-62.
• 11 Theoret JF, Bienvenu JG, Kumar A. et al. PSelectin antagonism with recombinant Pselectin glycoprotein ligand 1 (rPSGL-Ig) inhibits circulating activated platelet binding to neutrophils induced by damaged arterial surfaces. J Pharmacol Exp Ther 2001; 298: 658-64.
• 12 Phillips ML, Nudelman E, Gaeta FCA. et al. ELAM-1 Mediates cell adhesion by recognition of a carbohydrate ligand, Sialyl-Le^x . Science 1990; 250: 1130-2.
• 13 Erbe DV, Watson SR, Presta LG. et al. P-and E-selectin use common sites for carbohydrate ligand recognition and cell adhesion. J Cell Biol 1993; 120: 1227-35.
• 14 Kunz C, Rudloff S, Baier W. et al. Oligosaccharides in human milk: Structural, functional, and metabolic aspects. Annu Rev Nutr 2000; 20: 699-722.
• 15 Egge H. The diversity of oligosaccharides in human milk. In: New perspectives in infant nutrition. Renner Sawatzki. eds: Thieme, Stuttgart; New York: 1992: 12-26.
• 16 Engfer MB, Stahl B, Finke B. et al. Human milk oligosaccharides are resistant to enzymatic hydrolysis in the upper gastrointestinal tract. Am J Clin Nutr 2000; 71: 1589-96.
• 17 Gnoth MJ, Kunz C, Kinne-Saffran E. et al. Human milk oligosaccharides are minimally digested in vitro. J Nutr 2000; 130: 3014-20.
• 18 Gnoth MJ, Rudloff S, Kunz C. et al. Investigations of the in vitro transport of human milk oligosaccahrides by a Caco-2 monolayer using a novel high performance liquid chromatography-mass spectrometry technique. J Biol Chem 2001; 276: 34363-70.
• 19 Rudloff S, Pohlentz G, Diekmann L. et al. Urinary excretion of lactose and oligosaccharides in preterm infants fed human milk or infant formula. Acta Paediatr 1996; 85: 598-603.
• 20 Obermeier S, Rudloff S, Pohlentz G. et al. Secretion of 13C-labelled oligosaccharides into human milk and infant’s urine after an oral (13C)galactose load. Isotopes Environ Health Stud 1999; 35: 119-25.
• 21 Sharon N, Ofek I. Safe as mother’s milk: Carbohydrates as future anti-adhesion drugs for bacterial diseases. Glycoconj J 2000; 17: 659-64.
• 22 Kunz C, Rudloff S, Hintelmann A. et al. HighpH anion-exchange chromatography with pulsed amperometric detection and molar response factors of human milk oligosaccharides. J Chromatogr B 1996; 685: 211-21.
• 23 Gnoth MJ, Kunz C, Rudloff S. Endotoxinreduced milk oligosaccharide fractions suitable for cell biological studies. Eur J Med Res 2000; 05: 468-72.
• 24 Figdor CG, Leemans JM, Bont WS. et al. Theory and practice of centrifugal elutriation (CE). Factors influencing the separation of human blood cells. Cell Biophys 1983; 05: 105-18.
• 25 Jaffe EA, Nachman RL, Becker CG. et al. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest 1973; 52: 2745-56.
• 26 Mayer K, Merfels M, Muhly-Reinholz M. et al. Omega-3 Fatty acids suppress monocyte adhesion to human endothelial cells: role of endothelial PAF generation. Am J Physiol Heart Circ Physiol 2002; 283: H811-8.
• 27 Welply JK, Abbas SZ, Scudder P. et al. Multivalent sialyl-LeX: potent inhibitors of Eselectin-mediated cell adhesion; reagents for staining activated endothelial cells. Glycobiology 1994; 04: 259-65.
• 28 Rudloff S, Stefan C, Pohlentz G. et al. Detection of ligands for selectins in the oligosaccharide fraction of human milk. Eur J Nutr 2002; 41: 85-92.
• 29 Yoshida T, Toyama-Sorimachi N, Miyasaka M. et al. A crucial effect of ligand clustering on the inhibition of binding by L-selectin in intercellular adhesion. Biochem Biophys Res Commun 1994; 204: 969-74.
• 30 Maaheimo H, Renkonen R, Turunen JP. et al. Synthesis of a divalent sialyl Lewis x O-glycan, a potent inhibitor of lymphocyte-endothelium adhesion. Evidence that multivalency enhances the saccharide binding to L-selectin. Eur J Biochem 1995; 234: 616-25.
• 31 Zajchowski LD, Robbins SM. Lipid rafts and little caves. Compartmentalized signaling in membrane microdomains. Eur J Biochem 2002; 269: 737-52.
• 32 Kurzchalia TV, Parton RG. Membrane microdomains and caveolae. Curr Opin Cell Biol 1999; 11: 424-31.
• 33 Borges E, Eytner R, Moll T. et al. The P-selectin glycoprotein ligand-1 is important for recruitment of neutrophils into inflamed mouse peritoneum. Blood 1997; 90: 1934-42.
• 34 Etzioni A, Frydman M, Pollack S. et al. Brief report: recurrent severe infections caused by a novel leukocyte adhesion deficiency. N Engl J Med 1992; 327: 1789-92.
• 35 Musemeche C, Caplan M, Hsueh W. et al. Experimental necrotizing enterocolitis: the role of polymorphonuclear neutrophils. J Pediatr Surg 1991; 26: 1047-9.
• 36 Wright AL, Holberg CJ, Martinez FD. et al. Breast feeding and lower respiratory tract illness in the first year of life. Group Health Medical Associates. BMJ 1989; 299: 946-9.
• 37 Lucas A, Cole TJ. Breast milk and neonatal necrotising enterocolitis. Lancet 1990; 336: 1519-23.
• 38 Kosloske AM. Breast milk decreases the risk of neonatal necrotizing enterocolitis. Adv Nutr Res 2001; 10: 123-37.