Nucleotides (NT) and human milk oligosaccharides (HMO) individually affect epithelial cell growth, but their combined effects had not been studied. Herein, the impact of NT and HMO on cell proliferation, apoptosis, ne...Nucleotides (NT) and human milk oligosaccharides (HMO) individually affect epithelial cell growth, but their combined effects had not been studied. Herein, the impact of NT and HMO on cell proliferation, apoptosis, necrosis and cell cycle in the fetal epithelial cell line (FHs-74 Int) was determined. Cells were incubated with media containing 2.5% FBS and no epidermal growth factor (Control);fucosyllactose (FL) mix [85% 2’FL/15% 3’FL], sialyllactose (SL) mix [40% 6’SL/10% 3’SL/50% sialic acid (SA)] or LNnT at 125, 250, 500 or 1000 μg/mL with and without 250 μg/mL NT (43% CMP, 18.5% UMP, 16.4% AMP, and 22.0% GMP) for 24 or 72 h. NT alone significantly increased proliferation, but did not affect cell cycle or apoptosis/necrosis. All HMO treatments at 1000 μg/mL significantly decreased proliferation and some were also inhibitory at 250 or 500 μg/mL. When NT and HMO were simultaneously added, NT ameliorated the anti-proliferative effect of HMO. FL significantly increased cells in S phase and SL and LNnT treatments significantly increased cells in G2/M and S phases, which concomitantly decreased cells in G0/G1. HMO with NT significantly decreased the percent of cells in the G2/M phase compared to HMO alone. Higher HMO doses significantly increased the percentage of apoptotic and necrotic cells compared to control. In conclusion, HMO reduced cell proliferation and this effect is partially ameliorated by NT. It appears that HMO initially induced apoptosis/necrosis, which was later evidenced by G2/M cell cycle arrest and decreased proliferation.展开更多
Human milk oligosaccharides (HMO) contribute to innate immunity by enhancing growth of beneficial bacteria, epithelial cell maturation and mucosal barrier integrity. They have immunomodulatory effects and can block pa...Human milk oligosaccharides (HMO) contribute to innate immunity by enhancing growth of beneficial bacteria, epithelial cell maturation and mucosal barrier integrity. They have immunomodulatory effects and can block pathogen binding to host cell surface glycans or receptors. We investigated the effects of 2’-fucosyllactose (2’FL), 6’-sialyllactose (6’SL), 3’-sialyllactose (3’SL) and lacto-N-neoTetraose (LNnT) on human respiratory epithelial cell lines or peripheral blood mononuclear cells (PBMCs) following respiratory viral infectionin vitro. Expression of cytokines and viral load were monitored in infected cells. These biomarkers of innate immunity were selected since viral load and cytokine levels (IP-10, MIP-1α, IL-6, IL-8, TNF-α) have been correlated with disease severity in respiratory syncytial virus (RSV) and influenza (IAV) virus infectionin vivo. 2’FL significantly decreased RSV viral load and cytokines associated with disease severity (IL-6, IL-8, MIP-1α) and inflammation (TNF-α, MCP-1) in airway epithelial cells. LNnT and 6’SL significantly decreased IAV viral load in airway epithelial cells. 6’SL dose-dependently down-regulated IP-10 and TNF-α in RSV infected PBMCs. HMO at or below levels found in breast milk enhance innate immunity to respiratory viruses in vitro and may interact directly with cells to modulate biomarkers of innate immunity.展开更多
BACKGROUND Human milk oligosaccharides(HMOs)are bioactive components of breast milk with diverse health benefits,including shaping the gut microbiota,modulating the immune system,and protecting against infections.HMOs...BACKGROUND Human milk oligosaccharides(HMOs)are bioactive components of breast milk with diverse health benefits,including shaping the gut microbiota,modulating the immune system,and protecting against infections.HMOs exhibit dynamic secretion patterns during lactation,influenced by maternal genetics and environ-mental factors.Their direct and indirect antimicrobial properties have garnered significant research interest.However,a comprehensive understanding of the secretion dynamics of HMOs and their correlation with antimicrobial efficacy remains underexplored.AIM To synthesize current evidence on the secretion dynamics of HMOs during lactation and evaluate their antimicrobial roles against bacterial,viral,and protozoal pathogens.METHODS A systematic search of PubMed,Scopus,Web of Science,and Cochrane Library focused on studies investigating natural and synthetic HMOs,their secretion dynamics,and antimicrobial properties.Studies involving human,animal,and in vitro models were included.Data on HMO composition,temporal secretion patterns,and mechanisms of antimicrobial action were extracted.Quality assess-ment was performed using validated tools appropriate for study design.RESULTS A total of 44 studies were included,encompassing human,animal,and in vitro research.HMOs exhibited dynamic secretion patterns,with 2′-fucosyllactose(2′-FL)and lacto-N-tetraose peaking in early lactation and declining over time,while 3-fucosyllactose(3-FL)increased during later stages.HMOs demonstrated significant antimicrobial properties through pathogen adhesion inhibition,biofilm disruption,and enzymatic activity impairment.Synthetic HMOs,including bioengineered 2′-FL and 3-FL,were structurally and functionally comparable to natural HMOs,effectively inhibiting pathogens such as Pseudomonas aeruginosa,Escherichia coli,and Campylobacter jejuni.Additionally,HMOs exhibited synergistic effects with antibiotics,enhancing their efficacy against resistant pathogens.CONCLUSION HMOs are vital in antimicrobial defense,supporting infant health by targeting various pathogens.Both natural and synthetic HMOs hold significant potential for therapeutic applications,particularly in infant nutrition and as adjuncts to antibiotics.Further research,including clinical trials,is essential to address gaps in knowledge,validate findings,and explore the broader applicability of HMOs in improving maternal and neonatal health.展开更多
Human milk oligosaccharides(HMOs)are one of the major differences between livestock milk and human milk,and the prebiotic functions of HMOs have been verified through in vitro and clinical trials.The most abundant HMO...Human milk oligosaccharides(HMOs)are one of the major differences between livestock milk and human milk,and the prebiotic functions of HMOs have been verified through in vitro and clinical trials.The most abundant HMOs include 2′-fucysollactose(2′-FL),3-fucosyllactose(3-FL),lacto-N-neotetraose(LNnT)and lacto-N-tetraose(LNT);their application and synthesis have attracted wide attentions.In recent years,the biotechnological production of 2′-FL,3-FL,LNnT and LNT have emerged based on techniques such as whole-cell catalysis and fermentation.In particular,the development of metabolic engineering and synthetic biology methods and strategies have facilitated efficient biosynthesis of these HMOs.However,these advantages have not been systematically reviewed yet.In this review,we first discuss the structures and applications of HMOs;secondly,strategies of microbial synthesis of the most abundant 2′-FL,3-FL,LNnT and LNT are summarized and compared.Finally,challenges and perspectives of efficient microbial production of HMOs as well as strategies for overcoming the challenges are discussed.This review reveals the whole picture of recent development in HMOs microbial synthesis and can further facilitate the understanding of limiting factors,and further propose a few directions to promote the development of efficient production hosts.展开更多
文摘Nucleotides (NT) and human milk oligosaccharides (HMO) individually affect epithelial cell growth, but their combined effects had not been studied. Herein, the impact of NT and HMO on cell proliferation, apoptosis, necrosis and cell cycle in the fetal epithelial cell line (FHs-74 Int) was determined. Cells were incubated with media containing 2.5% FBS and no epidermal growth factor (Control);fucosyllactose (FL) mix [85% 2’FL/15% 3’FL], sialyllactose (SL) mix [40% 6’SL/10% 3’SL/50% sialic acid (SA)] or LNnT at 125, 250, 500 or 1000 μg/mL with and without 250 μg/mL NT (43% CMP, 18.5% UMP, 16.4% AMP, and 22.0% GMP) for 24 or 72 h. NT alone significantly increased proliferation, but did not affect cell cycle or apoptosis/necrosis. All HMO treatments at 1000 μg/mL significantly decreased proliferation and some were also inhibitory at 250 or 500 μg/mL. When NT and HMO were simultaneously added, NT ameliorated the anti-proliferative effect of HMO. FL significantly increased cells in S phase and SL and LNnT treatments significantly increased cells in G2/M and S phases, which concomitantly decreased cells in G0/G1. HMO with NT significantly decreased the percent of cells in the G2/M phase compared to HMO alone. Higher HMO doses significantly increased the percentage of apoptotic and necrotic cells compared to control. In conclusion, HMO reduced cell proliferation and this effect is partially ameliorated by NT. It appears that HMO initially induced apoptosis/necrosis, which was later evidenced by G2/M cell cycle arrest and decreased proliferation.
文摘Human milk oligosaccharides (HMO) contribute to innate immunity by enhancing growth of beneficial bacteria, epithelial cell maturation and mucosal barrier integrity. They have immunomodulatory effects and can block pathogen binding to host cell surface glycans or receptors. We investigated the effects of 2’-fucosyllactose (2’FL), 6’-sialyllactose (6’SL), 3’-sialyllactose (3’SL) and lacto-N-neoTetraose (LNnT) on human respiratory epithelial cell lines or peripheral blood mononuclear cells (PBMCs) following respiratory viral infectionin vitro. Expression of cytokines and viral load were monitored in infected cells. These biomarkers of innate immunity were selected since viral load and cytokine levels (IP-10, MIP-1α, IL-6, IL-8, TNF-α) have been correlated with disease severity in respiratory syncytial virus (RSV) and influenza (IAV) virus infectionin vivo. 2’FL significantly decreased RSV viral load and cytokines associated with disease severity (IL-6, IL-8, MIP-1α) and inflammation (TNF-α, MCP-1) in airway epithelial cells. LNnT and 6’SL significantly decreased IAV viral load in airway epithelial cells. 6’SL dose-dependently down-regulated IP-10 and TNF-α in RSV infected PBMCs. HMO at or below levels found in breast milk enhance innate immunity to respiratory viruses in vitro and may interact directly with cells to modulate biomarkers of innate immunity.
文摘BACKGROUND Human milk oligosaccharides(HMOs)are bioactive components of breast milk with diverse health benefits,including shaping the gut microbiota,modulating the immune system,and protecting against infections.HMOs exhibit dynamic secretion patterns during lactation,influenced by maternal genetics and environ-mental factors.Their direct and indirect antimicrobial properties have garnered significant research interest.However,a comprehensive understanding of the secretion dynamics of HMOs and their correlation with antimicrobial efficacy remains underexplored.AIM To synthesize current evidence on the secretion dynamics of HMOs during lactation and evaluate their antimicrobial roles against bacterial,viral,and protozoal pathogens.METHODS A systematic search of PubMed,Scopus,Web of Science,and Cochrane Library focused on studies investigating natural and synthetic HMOs,their secretion dynamics,and antimicrobial properties.Studies involving human,animal,and in vitro models were included.Data on HMO composition,temporal secretion patterns,and mechanisms of antimicrobial action were extracted.Quality assess-ment was performed using validated tools appropriate for study design.RESULTS A total of 44 studies were included,encompassing human,animal,and in vitro research.HMOs exhibited dynamic secretion patterns,with 2′-fucosyllactose(2′-FL)and lacto-N-tetraose peaking in early lactation and declining over time,while 3-fucosyllactose(3-FL)increased during later stages.HMOs demonstrated significant antimicrobial properties through pathogen adhesion inhibition,biofilm disruption,and enzymatic activity impairment.Synthetic HMOs,including bioengineered 2′-FL and 3-FL,were structurally and functionally comparable to natural HMOs,effectively inhibiting pathogens such as Pseudomonas aeruginosa,Escherichia coli,and Campylobacter jejuni.Additionally,HMOs exhibited synergistic effects with antibiotics,enhancing their efficacy against resistant pathogens.CONCLUSION HMOs are vital in antimicrobial defense,supporting infant health by targeting various pathogens.Both natural and synthetic HMOs hold significant potential for therapeutic applications,particularly in infant nutrition and as adjuncts to antibiotics.Further research,including clinical trials,is essential to address gaps in knowledge,validate findings,and explore the broader applicability of HMOs in improving maternal and neonatal health.
基金This work was financially supported by the National Natural Science Foundation of China(31871784,31870069,21676119 and 31671845)the National Key Research and Development Program of China(2018YFA0900300).
文摘Human milk oligosaccharides(HMOs)are one of the major differences between livestock milk and human milk,and the prebiotic functions of HMOs have been verified through in vitro and clinical trials.The most abundant HMOs include 2′-fucysollactose(2′-FL),3-fucosyllactose(3-FL),lacto-N-neotetraose(LNnT)and lacto-N-tetraose(LNT);their application and synthesis have attracted wide attentions.In recent years,the biotechnological production of 2′-FL,3-FL,LNnT and LNT have emerged based on techniques such as whole-cell catalysis and fermentation.In particular,the development of metabolic engineering and synthetic biology methods and strategies have facilitated efficient biosynthesis of these HMOs.However,these advantages have not been systematically reviewed yet.In this review,we first discuss the structures and applications of HMOs;secondly,strategies of microbial synthesis of the most abundant 2′-FL,3-FL,LNnT and LNT are summarized and compared.Finally,challenges and perspectives of efficient microbial production of HMOs as well as strategies for overcoming the challenges are discussed.This review reveals the whole picture of recent development in HMOs microbial synthesis and can further facilitate the understanding of limiting factors,and further propose a few directions to promote the development of efficient production hosts.
