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Published online before print July 7, 2004

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(Journal of Leukocyte Biology. 2004;76:820-826.)
© 2004 by Society for Leukocyte Biology

Human milk oligosaccharides reduce platelet-neutrophil complex formation leading to a decrease in neutrophil ß 2 integrin expression

Lars Bode^*,1, Silvia Rudloff^*, Clemens Kunz^*, Stephan Strobel and Nigel Klein

^* Institute of Nutritional Science, Justus-Liebig-University Giessen, Germany; and
Immunobiology Unit and
Infectious Diseases and Microbiology Unit, Institute of Child Health, University College London, United Kingdom

¹Correspondence at current address: The Burnham Institute, Glycobiology and Carbohydrate Chemistry Program, 10901 N. Torrey Pines Rd., La Jolla, CA 92037. E-mail: Lbode{at}burnham.org

Human milk is thought by many authorities to be preferable to formula as a source of nutrients for infants. Some of the benefits may stem from its high concentration of unbound oligosaccharides (5-10 g/L). These sugars have structural similarities to selectin ligands, known to mediate important cell–cell interactions in the immune system. Platelet-neutrophil complexes (PNC) exist in healthy individuals but have been implicated in disease states. Formation of these complexes requires selectins and as such, could be influenced by human milk oligosaccharides (HMO). Here, we investigate this possibility by examining the effect of HMO on the formation of PNC and activation of associated neutrophils. We collected blood from 10 healthy volunteers, activated platelets with adenosine 5'-diphosphate, and added HMO, oligosaccharide standards, or phosphate-buffered saline as a control. We determined the influence of HMO on PNC formation and adjacent neutrophil activation with fluorescein-activated cell sorter analysis after labeling with antibodies for the platelet marker CD42a and the neutrophil activation marker CD11b. Within physiologically achievable concentrations (6.25-125 µg/mL), an acidic HMO fraction reduced PNC formation up to 20%, which was similar to the effect seen with high concentrations of sialyl-Lewis x. Associated neutrophils showed a dose-dependent decrease in ß 2 integrin expression, up to 30%, at high but physiological concentrations. The neutral HMO fraction had no effect. These results support the hypothesis that acidic HMO serve as anti-inflammatory components of human milk and thus, contribute to the lower incidence of inflammatory diseases such as necrotizing enterocolitis in breast-fed versus formula-fed infants.

Key Words: breast-feeding • inflammation • platelets • neutrophils • selectins • integrins • necrotizing enterocolitis

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