Blends of Human Milk Oligosaccharides Confer Intestinal Epithelial Barrier Protection In Vitro

Breastfeeding is integral in the proper maturation of the intestinal barrier and protection against inflammatory diseases. When human milk (HM) is not available, supplementation with HM bioactives like Human Milk Oligosaccharides (HMOs) may help in providing breastfeeding barrier-protective benefits. An increasing HMO variety is becoming industrially available, enabling approaching the HMO complexity in HM. We aimed at assessing the impact of blends of available HMOs on epithelial barrier function in vitro. The capacity of individual [2′-Fucosyllactose (2′FL), Difucosyllactose, Lacto-N-tetraose, Lacto-N-neotetraose, 3′-Siallylactose and 6′-Siallylactose] or varying combinations of 3, 5 and 6 HMOs to modulate fluorescein-isothiocyanate (FITC)-labelled Dextran 4 KDa (FD4) translocation and/or transepithelial resistance (TEER) was characterized in Caco-2: HT29- methotrexate (MTX) cell line monolayers before and after an inflammatory challenge with TNF-α and IFN-γ. The six HMO blend (HMO6) dose-dependently limited the cytokine-induced FD4 translocation and TEER decrease and increased TEER values before challenge. Similarly, 3 and 5 HMO blends conferred a significant protection against the challenge, with 2′FL, one of the most abundant but most variable oligosaccharides in HM, being a key contributor. Overall, our results suggest differential ability of specific HMOs in modulating the intestinal barrier and support the potential of supplementation with combinations of available HMOs to promote gut health and protect against intestinal inflammatory disorders.

[1]  Mitchell R. Ladd,et al.  The human milk oligosaccharides 2’-Fucosyllactose and 6’-Sialyllactose protect against the development of necrotizing enterocolitis by inhibiting Toll-Like Receptor 4 signaling. , 2020, Pediatric Research.

[2]  M. Costeira,et al.  Impact of maternal characteristics on human milk oligosaccharide composition over the first 4 months of lactation in a cohort of healthy European mothers , 2019, Scientific Reports.

[3]  P. de Vos,et al.  Modulation of Intestinal Epithelial Glycocalyx Development by Human Milk Oligosaccharides and Non‐Digestible Carbohydrates , 2019, Molecular nutrition & food research.

[4]  H. Smidt,et al.  Correlating Infant Fecal Microbiota Composition and Human Milk Oligosaccharide Consumption by Microbiota of 1‐Month‐Old Breastfed Infants , 2019, Molecular nutrition & food research.

[5]  Bo Li,et al.  Human Milk Oligosaccharides Increase Mucin Expression in Experimental Necrotizing Enterocolitis , 2018, Molecular nutrition & food research.

[6]  J. Garssen,et al.  Diversity of Human Milk Oligosaccharides and Effects on Early Life Immune Development , 2018, Front. Pediatr..

[7]  L. Bode,et al.  Immunological Effects of Human Milk Oligosaccharides , 2018, Front. Pediatr..

[8]  L. Magder,et al.  Intestinal Barrier Maturation in Very Low Birthweight Infants: Relationship to Feeding and Antibiotic Exposure , 2017, The Journal of pediatrics.

[9]  James A. Foster,et al.  What’s normal? Oligosaccharide concentrations and profiles in milk produced by healthy women vary geographically12 , 2017, The American journal of clinical nutrition.

[10]  Hannah D. Holscher,et al.  Human Milk Oligosaccharides Influence Intestinal Epithelial Cell Maturation In Vitro , 2017, Journal of pediatric gastroenterology and nutrition.

[11]  N. Sprenger,et al.  FUT2-dependent breast milk oligosaccharides and allergy at 2 and 5 years of age in infants with high hereditary allergy risk , 2017, European Journal of Nutrition.

[12]  R. Haque,et al.  Fecal Markers of Environmental Enteropathy and Subsequent Growth in Bangladeshi Children. , 2016, The American journal of tropical medicine and hygiene.

[13]  H. Fukui.  Increased Intestinal Permeability and Decreased Barrier Function: Does It Really Influence the Risk of Inflammation? , 2016, Inflammatory Intestinal Diseases.

[14]  C. Lebrilla,et al.  Identification of Oligosaccharides in Feces of Breast-fed Infants and Their Correlation with the Gut Microbial Community * , 2016, Molecular & Cellular Proteomics.

[15]  A. Lowe,et al.  Breastfeeding and asthma and allergies: a systematic review and meta‐analysis , 2015, Acta paediatrica.

[16]  Nathan Lawlor,et al.  The human milk oligosaccharide 2′-fucosyllactose modulates CD14 expression in human enterocytes, thereby attenuating LPS-induced inflammation , 2014, Gut.

[17]  P. Denning,et al.  Human breast milk and the gastrointestinal innate immune system. , 2014, Clinics in perinatology.

[18]  Katherine Carroll,et al.  An exclusively human milk diet reduces necrotizing enterocolitis. , 2014, Breastfeeding medicine : the official journal of the Academy of Breastfeeding Medicine.

[19]  Hannah D. Holscher,et al.  Human milk oligosaccharides influence maturation of human intestinal Caco-2Bbe and HT-29 cell lines. , 2014, The Journal of nutrition.

[20]  Amy Raub,et al.  Breastfeeding policy: a globally comparative analysis. , 2013, Bulletin of the World Health Organization.

[21]  Lars Bode,et al.  Human milk oligosaccharides: every baby needs a sugar mama. , 2012, Glycobiology.

[22]  J. Sonnenburg,et al.  Human milk oligosaccharide consumption by intestinal microbiota. , 2012, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[23]  O. Gabrielli,et al.  Preterm Milk Oligosaccharides During the First Month of Lactation , 2011, Pediatrics.

[24]  J. Schrezenmeir,et al.  Prebiotic oligosaccharides reduce proinflammatory cytokines in intestinal Caco-2 cells via activation of PPARγ and peptidoglycan recognition protein 3. , 2011, The Journal of nutrition.

[25]  G. Williamson,et al.  Transport and Metabolism of Ferulic Acid through the Colonic Epithelium , 2008, Drug Metabolism and Disposition.

[26]  Rudolf Grimm,et al.  A strategy for annotating the human milk glycome. , 2006, Journal of agricultural and food chemistry.

[27]  S. Reif,et al.  Breastfeeding and risk of inflammatory bowel disease: a systematic review with meta-analysis. , 2004, The American journal of clinical nutrition.

[28]  J. Meinzen-Derr,et al.  Innate protection conferred by fucosylated oligosaccharides of human milk against diarrhea in breastfed infants. , 2004, Glycobiology.

[29]  G. Coppa,et al.  Intestinal Permeability. Changes during the First Month: Effect of Natural versus Artificial Feeding , 1995, Journal of pediatric gastroenterology and nutrition.

[30]  I. Jakobssonc.  Intestinal permeability in children of different ages and with different gastrointestinal diseases , 1993 .

[31]  I. Jakobsson.  Intestinal permeability in children of different ages and with different gastrointestinal diseases. , 1993, Pediatric allergy and immunology : official publication of the European Society of Pediatric Allergy and Immunology.