Lactodifucotetraose, a human milk oligosaccharide, attenuates platelet function and inflammatory cytokine release


Human milk strongly quenches inflammatory processes in vitro, and breastfed infants have lower incidence of inflammatory diseases than those fed artificially. Platelets from neonates, in contrast to those from adults, are less responsive to platelet agonists such as collagen, thrombin, ADP, and epinephrine. Breastfed infants absorb oligosaccharides intact from the human milk in their gut to the circulation. This study was to determine whether these oligosaccharides can attenuate platelet function and platelet secretion of pro-inflammatory proteins, and to identify the active component. The natural mixture of oligosaccharides from human milk and pure individual human milk oligosaccharides were tested for their ability to modulate responses of platelets isolated from human blood following exposure to thrombin, ADP, and collagen. Human milk and the natural mixture of human milk oligosaccharides inhibited platelet release of inflammatory proteins. Of the purified human milk oligosaccharides tested, only lactodifucotetraose (LDFT) significantly inhibited thrombin induced release of the pro-inflammatory proteins RANTES and sCD40L. LDFT also inhibited platelet adhesion to a collagen-coated surface, as well as platelet aggregation induced by ADP or collagen. These data indicate that LDFT may help modulate hemostasis by suppressing platelet-induced inflammatory processes in breastfed infants. This activity suggests further study of LDFT for its potential as a therapeutic agent in infants and adults.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4


  1. 1.

    Nanthakumar NN, Meng D, Newburg DS (2013) Glucocorticoids and microbiota regulate ontogeny of intestinal fucosyltransferase 2 requisite for gut homeostasis. Glycobiology 23(10):1131–1141. doi:10.1093/glycob/cwt050

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  2. 2.

    Meng D, Newburg DS, Young C, Baker A, Tonkonogy SL, Sartor RB, Walker WA, Nanthakumar NN (2007) Bacterial symbionts induce a FUT2-dependent fucosylated niche on colonic epithelium via ERK and JNK signaling. Am J Physiol Gastrointest Liver Physiol 293(4):G780–G787. doi:10.1152/ajpgi.00010.2007

    CAS  Article  PubMed  Google Scholar 

  3. 3.

    He Y, Liu S, Leone S, Newburg DS (2014) Human colostrum oligosaccharides modulate major immunologic pathways of immature human intestine. Mucosal Immunol 7(6):1326–1339. doi:10.1038/mi.2014.20

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  4. 4.

    He Y, Liu S, Kling DE, Leone S, Lawlor NT, Huang Y, Feinberg SB, Hill DR, Newburg DS (2014) The human milk oligosaccharide 2′-fucosyllactose modulates CD14 expression in human enterocytes, thereby attenuating LPS-induced inflammation. Gut. doi:10.1136/gutjnl-2014-307544

    Google Scholar 

  5. 5.

    Goehring KC, Kennedy AD, Prieto PA, Buck RH (2014) Direct evidence for the presence of human milk oligosaccharides in the circulation of breastfed infants. PLoS One 9(7):1–11. doi:10.1371/journal.pone.0101692

    Article  Google Scholar 

  6. 6.

    Chaturvedi P, Warren CD, Altaye M, Morrow AL, Ruiz-Palacios GM, Pickering LK, Newburg DS(2001) Fucosylated human milk oligosaccharides vary among individuals over the course of lactation. Glycobiology 365-72. doi: 10.1093/glycob/11.5.365

  7. 7.

    Chaturvedi P, Warren CD, Buescher CR, Pickering LK, Newburg DS (2001) Survival of human milk oligosaccharides in the intestine of infants. Adv Exp Med Biol 501:315–323

    CAS  Article  PubMed  Google Scholar 

  8. 8.

    Ruhaak R, Stroble C, Underwood MA, Lebrilla CD (2014) Detection of milk oligosaccharides in plasma of infants. Anal Bioanal Chem 406(24):5775–5784. doi:10.1007/s00216-014-8025-z

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  9. 9.

    Bode L, Kunz C, Muhly-Reinholz M, Mayer K, Seeger W, Rudloff S (2004) Inhibition of monocyte, lymphocyte, and neutrophil adhesion to endothelial cells by human milk oligosaccharides. Thromb Haemost 92(6):1402–1410. doi:10.1267/THRO04061402

    CAS  PubMed  Google Scholar 

  10. 10.

    Zarbock A, Polanowska-Grabowska RK, Ley K (2007) Platelet-neutrophil-interactions: linking hemostasis and inflammation. Blood Rev 21(2):99–111. doi:10.1016/j.blre.2006.06.001

    CAS  Article  PubMed  Google Scholar 

  11. 11.

    Wagner DD, Burger PC (2003) Platelets in inflammation and thrombosis. Arterioscler Thromb Vasc Biol 23(12):2131–2137. doi:10.1161/01.ATV.0000095974.95122.EC

    CAS  Article  PubMed  Google Scholar 

  12. 12.

    Bode L, Rudloff S, Kunz C, Strobel S, Klein N (2004) Human milk oligosaccharides reduce platelet-neutrophil complex formation leading to a decrease in neutrophil beta 2 integrin expression. J Leukoc Biol 76(4):820–826. doi:10.1189/jlb.0304198

    CAS  Article  PubMed  Google Scholar 

  13. 13.

    Coppinger JA, Cagney G, Toomey S, Kislinger T, Belton O, McRedmond JP, Cahill DJ, Emili A, Fitzgerald DJ, Maguire PB (2004) Characterization of the proteins released from activated platelets leads to localization of novel platelet proteins in human atherosclerotic lesions. Blood 103(6):2096–2104. doi:10.1182/blood-2003-08-2804

    CAS  Article  PubMed  Google Scholar 

  14. 14.

    Danese S, Fiocchi C (2003) Atherosclerosis and inflammatory bowel disease: sharing a common pathogenic pathway? Circulation 107(7):e5211

    Article  Google Scholar 

  15. 15.

    Aukrust P, Ueland T, Muller F, Andreassen AK, Nordoy I, Aas H, Kjekshus J, Simonsen S, Froland SS, Gullestad L (1998) Elevated circulating levels of C-C chemokines in patients with congestive heart failure. Circulation 97(12):1136–1143

    CAS  Article  PubMed  Google Scholar 

  16. 16.

    Danese S, de la Motte C, Sturm A, Vogel JD, West GA, Strong SA, Katz JA, Fiocchi C (2003) Platelets trigger a CD40-dependent inflammatory response in the microvasculature of inflammatory bowel disease patients. Gastroenterology 124(5):1249–1264

    CAS  Article  PubMed  Google Scholar 

  17. 17.

    Newburg DS, Pickering LK, McCluer RH, Cleary TG (1990) Fucosylated oligosaccharides of human milk protect suckling mice from heat-stabile enterotoxin of Escherichia coli. J Infect Dis 162:1075–1080

    CAS  Article  PubMed  Google Scholar 

  18. 18.

    Chakrabarti S, Varghese S, Vitseva O, Tanriverdi K, Freedman JE (2005) CD40 ligand influences platelet release of reactive oxygen intermediates. Arterioscler Thromb Vasc Biol 25(11):2428–2434. doi:10.1161/01.ATV.0000184765.59207.f3

    CAS  Article  PubMed  Google Scholar 

  19. 19.

    Chakrabarti S, Vitseva O, Iyu D, Varghese S, Freedman JE (2005) The effect of dipyridamole on vascular cell-derived reactive oxygen species. J Pharmacol Exp Ther 315(2):494–500. doi:10.1124/jpet.105.089987

    CAS  Article  PubMed  Google Scholar 

  20. 20.

    Vanichakarn P, Blair P, Wu C, Freedman JE, Chakrabarti S (2008) Neutrophil CD40 enhances platelet-mediated inflammation. Thromb Res 122(3):346–358. doi:10.1016/j.thromres.2007.12.019

    CAS  Article  PubMed  Google Scholar 

  21. 21.

    Yu ZT, Chen C, Newburg DS (2013) Utilization of major fucosylated and sialylated human milk oligosaccharides by isolated human gut microbes. Glycobiology 23(11):1205–1206. doi:10.1093/glycob/cws196

    Article  Google Scholar 

  22. 22.

    Newburg DS (2013) Glycobiology of human milk. Biochemistry (Mosc) 78(7):771–785. doi:10.1134/S0006297913070092

    CAS  Article  Google Scholar 

  23. 23.

    Giglia TM, Massicotte MP, Tweddell JS, Barst RJ, Bauman M, Erickson CC, Feltes TF, Foster E, Hinoki K, Ichord RN, Kreutzer J, McCrindle BW, Newburger JW, Tabbutt S, Todd JL, Webb CL, American Heart Association Congenital Heart Defects Committee of the Council on Cardiovascular Disease in the Young CoC, Stroke Nursing CoE, Prevention, Stroke C (2013) Prevention and treatment of thrombosis in pediatric and congenital heart disease: a scientific statement from the American Heart Association. Circulation 128(24):2622–2703. doi:10.1161/01.cir.0000436140.77832.7a

    Article  PubMed  Google Scholar 

  24. 24.

    Morrow AL, Ruiz-Palacios GM, Altaye M, Jiang X, Guerrero ML, Meinzen-Derr JK, Farkas T, Chaturvedi P, Pickering LK, Newburg DS (2004) Human milk oligosaccharides are associated with protection against diarrhea in breast-fed infants. J Pediatr 145(3):297–303. doi:10.1016/j.jpeds.2004.04.054

    CAS  Article  PubMed  Google Scholar 

  25. 25.

    Westerbeek EAM, van dem Berg A, Lafeber HN, Fetter WPF, van Elburg RM (2011) The effect of enteral supplementation of a prebiotic mixture of non-human milk galacto-, fructo-, and acidic oligosaccharides on intestinal permeability in preterm infants. Br J Nutr 105:268–274. doi:10.1017/S0007114510003405

    CAS  Article  PubMed  Google Scholar 

  26. 26.

    Andre P, Nannizzi-Alaimo L, Prasad SK, Phillips DR (2002) Platelet-derived CD40L: the switch-hitting player of cardiovascular disease. Circulation 106(8):896–899

    Article  PubMed  Google Scholar 

  27. 27.

    Inwald DP, McDowall A, Peters MJ, Callard RE, Klein NJ (2003) CD40 is constitutively expressed on platelets and provides a novel mechanism for platelet activation. Circ Res 92(9):1041–1048. doi:10.1161/01.RES.0000070111.98158.6C

    CAS  Article  PubMed  Google Scholar 

  28. 28.

    Chakrabarti S, Blair P, Freedman JE (2007) CD40-40L signaling in vascular inflammation. J Biol Chem 282(25):18307–18317. doi:10.1074/jbc.M700211200

    CAS  Article  PubMed  Google Scholar 

  29. 29.

    Andre P, Prasad KS, Denis CV, He M, Papalia JM, Hynes RO, Phillips DR, Wagner DD (2002) CD40L stabilizes arterial thrombi by a beta3 integrin–dependent mechanism. Nat Med 8(3):247–252. doi:10.1038/nm0302-247

    CAS  Article  PubMed  Google Scholar 

  30. 30.

    Danese S, de la Motte C, Reyes BM, Sans M, Levine AD, Fiocchi C (2004) Cutting edge: T cells trigger CD40-dependent platelet activation and granular RANTES release: a novel pathway for immune response amplification. J Immunol 172(4):2011–2015

    CAS  Article  PubMed  Google Scholar 

  31. 31.

    Kaplan ZS, Jackson SP (2011) The role of platelets in atherothrombosis. Hematology Am Soc Hematol Educ Program 2011:51–61. doi:10.1182/asheducation-2011.1.51

    PubMed  Google Scholar 

  32. 32.

    Woulfe D, Yang J, Brass L (2001) ADP and platelets: the end of the beginning. J Clin Invest 107(12):1503–1505. doi:10.1172/JCI13361

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  33. 33.

    Pugh N, Simpson AM, Smethurst PA, de Groot PG, Raynal N, Farndale RW (2010) Synergism between platelet collagen receptors defined using receptor-specific collagen-mimetic peptide substrata in flowing blood. Blood 115(24):5069–5079. doi:10.1182/blood-2010-01-260778

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  34. 34.

    Coughlin SR (2000) Thrombin signalling and protease-activated receptors. Nature 407(6801):258–264. doi:10.1038/35025229

    CAS  Article  PubMed  Google Scholar 

  35. 35.

    Rizvi M, Pathak D, Freedman JE, Chakrabarti S (2008) CD40-CD40 ligand interactions in oxidative stress, inflammation and vascular disease. Trends Mol Med 14(12):530–538. doi:10.1016/j.molmed.2008.09.006

    CAS  Article  PubMed  Google Scholar 

  36. 36.

    Ignjatovic V, Kenet G, Monagle P, Perinatal, Paediatric Haemostasis Subcommittee of the S, Standardization Committee of the International Society on T, Haemostasis (2012) Developmental hemostasis: recommendations for laboratories reporting pediatric samples. J Thromb Haemost 10(2):298–300

    CAS  Article  PubMed  Google Scholar 

Download references


Supported in part by the National Institute of Allergy and Infectious Diseases AI075563, Eunice Kennedy Shriver National Institute of Child Health and Human Development HD013021 and HD059140. The authors appreciate the assistance of Dr. N. Nanda Nanthakumar in the design of the figures.

Author information



Corresponding author

Correspondence to David S. Newburg.

Ethics declarations

Conflict of interest

DSN owns stock in Glycosyn, LLC, which makes human milk oligosaccharides. This competition of interests is being managed by Boston College. The other authors have no competing interests to disclose.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Newburg, D.S., Tanritanir, A.C. & Chakrabarti, S. Lactodifucotetraose, a human milk oligosaccharide, attenuates platelet function and inflammatory cytokine release. J Thromb Thrombolysis 42, 46–55 (2016).

Download citation


  • Human milk oligosaccharides
  • Lactodifucotetraose
  • Platelets
  • Inflammation
  • Aggregation