Inflammatory response according to diets.

<p>Inflammatory response assessed in the BALF of the pAOS and control mice during the first 4 days of PA infection. <i>A</i>, number of neutrophils. <i>B</i>, number of macrophages. <i>C</i>, KC concentration. <i>D</i>, TNF-α concentration. <i>E</i>, IL-10 concentration. Each open circle represents one mouse fed the control diet; each black circle represents one mouse fed the pAOS diet. Bars represent the median values. Dotted lines indicate differences between each day of infection within each dietary mice group; solid lines indicate differences between the pAOS and control mice groups on the same day. the p-values are based on the Mann-Whitnney U test. *, <i>P</i><0.05; **, <i>P</i><0.01.</p

Survival rates according to diets.

<p>Survival rates after PA infection in mice fed the pAOS or control diet. The p-value is based on the log-rank test. *, <i>P</i><0.05.</p

concentrations of SCFAs according diets.

<p>Data were analysed using the Mann-Whitney<i>U</i> test.</p><p>*<i>P</i><0.05.</p><p>Concentrations (μg/100mg) of acetic acid (C2:0), propionic acid (C3:0), isobutyric acid (C4:i), butyric acid (C4:0), isovaleric acid (C5:0i) and valeric acid (C5) in feces of mice fed the control or pAOS diet at different times during the dietary intervention.</p

Immune response according to diets.

<p>Immune response assessed in the lungs of mice fed the pAOS or control diet during the first 4 days after PA infection. <i>A</i>, IFN-γ IL-4 concentrations and IFN- γ/IL-4 ratio. <i>B</i>, <i>nos2</i>, <i>arg1</i>, and <i>nos2/arg1</i> mRNA levels. Each open circle represents one mouse fed control diet and each black circle represents one mouse fed pAOS diet. Bars represent median values. Lines indicate differences between each day of infection within each dietary mice group or between the pAOS and control mice groups on the same day of infection.; The p-values are based on the Mann-Whitney U test. *, <i>P</i><0.05; **, <i>P</i><0.01</p

Bacterial load in lungs according to diets.

<p>Bacterial load in lungs assessed as the ratio of the number of bacteria recovered from each mouse and the number of bacteria injected after a first PA infection. Each open circle represents one mouse fed the control diet; each black circle represents one mouse fed the pAOS diet. Bars represent median values. Dotted lines indicate differences between each day of infection within each dietary mice group; solid lines indicate differences between the pAOS and control mice groups on the same day; The p-values are based on the MannWhitney U test. *, <i>P</i><0.05; **, <i>P</i><0.01.</p

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Background<p>Human milk is uniquely suited to provide optimal nutrition and immune protection to infants. Human milk oligosaccharides are structural complex and diverse consisting of short chain and long chain oligosaccharides typically present in a 9:1 ratio. 2′-Fucosyllactose (2′FL) is one of the most prominent short chain oligosaccharides and is associated with anti-infective capacity of human milk.</p>Aim<p>To determine the effect of 2′FL on vaccination responsiveness (both innate and adaptive) in a murine influenza vaccination model and elucidate mechanisms involved.</p>Methods<p>A dose range of 0.25–5% (w/w) dietary 2′FL was provided to 6-week-old female C57Bl/6JOlaHsd mice 2 weeks prior primary and booster vaccination until the end of the experiment. Intradermal (i.d.) challenge was performed to measure the vaccine-specific delayed-type hypersensitivity (DTH). Antigen-specific antibody levels in serum as well as immune cell populations within several organs were evaluated using ELISA and flow cytometry, respectively. In an ex vivo restimulation assay, spleen cells were cocultured with influenza-loaded bone marrow-derived dendritic cells (BMDCs) to study the effects of 2′FL on vaccine-specific CD4+ and CD8+ T-cell proliferation and cytokine secretions. Furthermore, the direct immune regulatory effects of 2′FL were confirmed using in vitro BMDCs T-cell cocultures.</p>Results<p>Dietary 2′FL significantly (p < 0.05) enhanced vaccine specific DTH responses accompanied by increased serum levels of vaccine-specific immunoglobulin (Ig) G1 and IgG2a in a dose-dependent manner. Consistently, increased activation marker (CD27) expression on splenic B-cells was detected in mice receiving 2′FL as compared to control mice. Moreover, proliferation of vaccine-specific CD4+ and CD8+ T-cells, as well as interferon-γ production after ex vivo restimulation were significantly increased in spleen cells of mice receiving 2′FL as compared to control mice, which were in line with changes detected within dendritic cell populations. Finally, we confirmed a direct effect of 2′FL on the maturation status and antigen presenting capacity of BMDCs.</p>Conclusion<p>Dietary intervention with 2′FL improves both humoral and cellular immune responses to vaccination in mice, which might be attributed in part to the direct effects of 2′FL on immune cell differentiation.</p

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Background<p>Human milk is uniquely suited to provide optimal nutrition and immune protection to infants. Human milk oligosaccharides are structural complex and diverse consisting of short chain and long chain oligosaccharides typically present in a 9:1 ratio. 2′-Fucosyllactose (2′FL) is one of the most prominent short chain oligosaccharides and is associated with anti-infective capacity of human milk.</p>Aim<p>To determine the effect of 2′FL on vaccination responsiveness (both innate and adaptive) in a murine influenza vaccination model and elucidate mechanisms involved.</p>Methods<p>A dose range of 0.25–5% (w/w) dietary 2′FL was provided to 6-week-old female C57Bl/6JOlaHsd mice 2 weeks prior primary and booster vaccination until the end of the experiment. Intradermal (i.d.) challenge was performed to measure the vaccine-specific delayed-type hypersensitivity (DTH). Antigen-specific antibody levels in serum as well as immune cell populations within several organs were evaluated using ELISA and flow cytometry, respectively. In an ex vivo restimulation assay, spleen cells were cocultured with influenza-loaded bone marrow-derived dendritic cells (BMDCs) to study the effects of 2′FL on vaccine-specific CD4+ and CD8+ T-cell proliferation and cytokine secretions. Furthermore, the direct immune regulatory effects of 2′FL were confirmed using in vitro BMDCs T-cell cocultures.</p>Results<p>Dietary 2′FL significantly (p < 0.05) enhanced vaccine specific DTH responses accompanied by increased serum levels of vaccine-specific immunoglobulin (Ig) G1 and IgG2a in a dose-dependent manner. Consistently, increased activation marker (CD27) expression on splenic B-cells was detected in mice receiving 2′FL as compared to control mice. Moreover, proliferation of vaccine-specific CD4+ and CD8+ T-cells, as well as interferon-γ production after ex vivo restimulation were significantly increased in spleen cells of mice receiving 2′FL as compared to control mice, which were in line with changes detected within dendritic cell populations. Finally, we confirmed a direct effect of 2′FL on the maturation status and antigen presenting capacity of BMDCs.</p>Conclusion<p>Dietary intervention with 2′FL improves both humoral and cellular immune responses to vaccination in mice, which might be attributed in part to the direct effects of 2′FL on immune cell differentiation.</p

Data_Sheet_1_Human milk nutritional composition across lactational stages in Central Africa.docx

The African region encompasses the highest undernutrition burden with the highest neonatal and infant mortality rates globally. Under these circumstances, breastfeeding is one of the most effective ways to ensure child health and development. However, evidence on human milk (HM) composition from African women is scarce. This is of special concern, as we have no reference data from HM composition in the context of food insecurity in Africa. Furthermore, data on the evolution of HM across lactational stages in this setting lack as well. In the MITICA study, we conducted a cohort study among 48 Central-African women and their 50 infants to analyze the emergence of gut dysbiosis in infants and describe the mother-infant transmission of microbiota between birth and 6 months of age. In this context, we assessed nutritional components in HM of 48 lactating women in Central Africa through five sampling times from week 1 after birth until week 25. Unexpectedly, HM-type III (Secretor + and Lewis genes -) was predominant in HM from Central African women, and some nutrients differed significantly among HM-types. While lactose concentration increased across lactation periods, fatty acid concentration did not vary significantly. The overall median level of 16 detected individual human milk oligosaccharides (HMOs; core structures as well as fucosylated and sialylated ones) decreased from 7.3 g/l at week 1 to 3.5 g/l at week 25. The median levels of total amino acids in HM dropped from 12.8 mg/ml at week 1 to 7.4 mg/ml at week 25. In contrast, specific free amino acids increased between months 1 and 3 of lactation, e.g., free glutamic acid, glutamine, aspartic acid, and serine. In conclusion, HM-type distribution and certain nutrients differed from Western mother HM.</p