Acidified dairy products are one of the oldest types of food products. Unfortunately all of them are low in dietary fibre. Thus, to improve health benefit of these products the idea of fortifying them with dietary fibre seems attractive. However dairy products enriched with Glucagel (a commercial product that is high in barley β-glucan) were found to suffer from textural defects. When the Glucagel concentration exceeded a certain value (5 g/L), dramatic phase separation was observed in set yogurt and yogurt drink with volume fraction of casein micelles greater then 0.108. To investigate interactions of β-glucan polymers and casein micelles in the milk prior to setting of yogurt, mixtures of yogurt milk and Glucagel were systematically studied. Depending on the volume fraction of casein micelles and the Glucagel concentration, a stable phase or a gel or a sedimented material could exist. The driving force for phase separation was depletion flocculation of casein micelles in the presence of β-glucan. The phase separation responsible for textural defects in yogurt systems supplemented with high amounts of Glucagel can be avoided by the reduction of β-glucan molecular weight, a process that limits the range of attraction between micelles. Incubation of Glucagel with lichenase for 90 min resulted in homogeneous (stable) yogurt systems with Glucagel concentrations as high as 10 g/L.

Acidified dairy products are one of the oldest types of food products. Unfortunately all of them are low in dietary fibre. Thus, to improve health benefit of these products the idea of fortifying them with dietary fibre seems attractive. However dairy products enriched with Glucagel (a commercial product that is high in barley β-glucan) were found to suffer from textural defects. When the Glucagel concentration exceeded a certain value (5 g/L), dramatic phase separation was observed in set yogurt and yogurt drink with volume fraction of casein micelles greater then 0.108. To investigate interactions of β-glucan polymers and casein micelles in the milk prior to setting of yogurt, mixtures of yogurt milk and Glucagel were systematically studied. Depending on the volume fraction of casein micelles and the Glucagel concentration, a stable phase or a gel or a sedimented material could exist. The driving force for phase separation was depletion flocculation of casein micelles in the presence of β-glucan. The phase separation responsible for textural defects in yogurt systems supplemented with high amounts of Glucagel can be avoided by the reduction of β-glucan molecular weight, a process that limits the range of attraction between micelles. Incubation of Glucagel with lichenase for 90 min resulted in homogeneous (stable) yogurt systems with Glucagel concentrations as high as 10 g/L.

Background: Candida albicans is an opportunistic fungal pathogen that induces strong proinflammatory responses, such as IL-1β production. Much less is known about the induction of immune modulatory cytokines, such as the IL-1 receptor antagonist (IL-1Ra) that is the main natural antagonist of IL-1, by C. albicans. Methods: Peripheral blood mononuclear cells (PBMC) of healthy individuals were stimulated with C. albicans and different components of the fungal cell wall. The role of pathogen recognition receptors (PRRs) for the induction of IL-1β and IL-1Ra was investigated by using specific blockers or in PBMC from Dectin-1 deficient patients. Results: C. albicans induced a strong IL-1Ra response, and this induction was primarily induced by the cell-wall component β-glucan. Blocking IL-1Ra significantly increased C. albicans β-glucan hyphae induced IL-1β and IL-6 production. Surprisingly, blocking the β-glucan receptor Dectin-1 or the downstream Syk or Raf-1 pathways only marginally reduced C. albicans-induced IL-1Ra production, while blocking of the complement receptor 3 (CR3), TLR2 or TLR4 had no effect. In line with this, blocking MAP kinases had little effect on Candida-induced IL-1Ra production. PBMC isolated from Dectin-1 deficient patients produced normal IL-1Ra amounts in response to C. albicans stimulation. Interestingly, the IL-1Ra synthesis induced by β-glucan was blocked by inhibitors of the Akt/PI3. K pathway. Conclusions: β-glucan of C. albicans induces a strong IL-1Ra response, which is independent of the β-glucan receptors dectin-1 and CR3. These data strongly argue for the existence of an unknown β-glucan receptor that specifically induces an Akt/PI3. K-dependent anti-inflammatory IL-1Ra response upon recognition of C. albicans.

Recognition of the fungal cell wall carbohydrate β-glucan by the host receptor Dectin-1 elicits broad immunomodulatory responses, such as phagocytosis and activation of oxidative burst. These responses are essential for engulfing and killing fungal pathogens. Phagocytic monocytes are key mediators of these early host inflammatory responses to infection. Remarkably, whether phagocytosis of fungal β-glucan leads to an inflammatory response in human monocytes remains to be established. Here, we show that phagocytosis of heat-killed Candida albicans is essential to trigger inflammation and cytokine release. By contrast, inhibition of actin-dependent phagocytosis of particulate (1-3,1-6)-β-glucan induces a strong inflammatory signature. Sustained monocyte activation, induced by fungal β-glucan particles upon actin cytoskeleton disruption, relies on Dectin-1 and results in the classical caspase-1 inflammasome formation through NLRP3, generation of an oxidative burst, NF-κB activation, and increased inflammatory cytokine release. PI3K and NADPH oxidase were crucial for both cytokine secretion and ROS generation, whereas Syk signaling mediated only cytokine production. Our results highlight the mechanism by which phagocytosis tightly controls the activation of phagocytes by fungal pathogens and strongly suggest that actin cytoskeleton dynamics are an essential determinant of the host's susceptibility or resistance to invasive fungal infections.

Escalating consumer concerns have placed the poultry industry under mounting pressure to reduce the use of chemotherapeutic agents as feed additives. One possible alternative receiving increased attention is the use of immunomodulators such as β-glucan. A pilot study evaluated the effects of a yeast derived β-glucan (Auxoferm YGT) on growth performance and immune response of broiler chickens. Day-old chicks were fed a diet containing 0, 0.02, or 0.1% yeast β-glucan. On days 7 and 14 post-hatch, body weight and relative immune organ weights were measured, peripheral blood was collected to determine heterophil to lymphocyte (H:L) ratios, and small intestinal sections were sampled to evaluate relative gene expression. The addition of β-glucan had no influence on growth. Dietary β-glucan supplementation modulated the expression of interleukin (IL)-8, IL-18, interferon (IFN)-γ and inducible nitric oxide synthase (iNOS) in the small intestine. A subsequent study was conducted to investigate the effects of dietary β-glucan on broiler chick (1440 birds) performance and immune response during a mixed Eimeria infection (day 8 of age). Measurements were taken and samples collected on days 4, 7, 10, 14 and 21 post-hatch. The results from this study show that β-glucan supplementation did not negatively impact performance. The addition of β-glucan to the diet resulted in reduced gross lesion severity and increased H:L ratios. The gene expression results suggest that β-glucans are capable of skewing the host immune response toward aTh1 mediated response and consequently down-regulating the Th2 mediated response. / Master of Science

Microglia, the innate immune cells of the central nervous system, feature adaptive immune memory with implications for brain homeostasis and pathologies. However, factors involved in the emergence and regulation of these opposing responses in microglia have not been fully addressed. Recently, we showed that microglia from the newborn brain display features of trained immunity and immune tolerance after repeated contact with pathogens in a dose-dependent manner. Here, we evaluate the impact of developmental stage on adaptive immune responses of brain microglia after repeated challenge with ultra-low (1 fg/ml) and high (100 ng/ml) doses of the endotoxin LPS in vitro. We find that priming of naïve microglia derived from newborn but not mature and aged murine brain with ultra-low LPS significantly increased levels of pro-inflammatory mediators TNF-α, IL-6, IL-1β, MMP-9, and iNOS as well as neurotrophic factors indicating induction of trained immunity (p < 0.05). In contrast, stimulation with high doses of LPS led to a robust downregulation of pro-inflammatory cytokines and iNOS independent of the developmental state, indicating induced immune tolerance. Furthermore, high-dose priming with LPS upregulated anti-inflammatory mediators IL-10, Arg-1, TGF- β, MSR1, and IL-4 in newborn microglia (p < 0.05). Our data indicate pronounced plasticity of the immune response of neonate microglia compared with microglia derived from mature and aged mouse brain. Induced trained immunity after priming with ultra-low LPS doses may be responsible for enhanced neuro-inflammatory susceptibility of immature brain. In contrast, the immunosuppressed phenotype following high-dose LPS priming might be prone to attenuate excessive damage after recurrent systemic inflammation.

Trained immunity and immune tolerance have been identified as long-term response patterns of the innate immune system. The causes of these opposing reactions remain elusive. Here, we report about differential inflammatory responses of microglial cells derived from neonatal mouse brain to increasing doses of the endotoxin LPS. Prolonged priming with ultra-low LPS doses provokes trained immunity, i.e., increased production of pro-inflammatory mediators in comparison to the unprimed control. In contrast, priming with high doses of LPS induces immune tolerance, implying decreased production of inflammatory mediators and pronounced release of anti-inflammatory cytokines. Investigation of the signaling processes and cell functions involved in these memory-like immune responses reveals the essential role of phosphoinositide 3-kinase γ (PI3Kγ), one of the phosphoinositide 3-kinase species highly expressed in innate immune cells. Together, our data suggest profound influence of preceding contacts with pathogens on the immune response of microglia. The impact of these interactions—trained immunity or immune tolerance—appears to be shaped by pathogen dose.

The ability of fungal-derived β-glucan particles to induce leukocyte activation and the production of inflammatory mediators, such as tumor necrosis factor (TNF)-α, is a well characterized phenomenon. Although efforts have been made to understand how these carbohydrate polymers exert their immunomodulatory effects, the receptors involved in generating these responses are unknown. Here we show that Dectin-1 mediates the production of TNF-α in response to zymosan and live fungal pathogens, an activity that occurs at the cell surface and requires the cytoplasmic tail and immunoreceptor tyrosine activation motif of Dectin-1 as well as Toll-like receptor (TLR)-2 and Myd88. This is the first demonstration that the inflammatory response to pathogens requires recognition by a specific receptor in addition to the TLRs. Furthermore, these studies implicate Dectin-1 in the production of TNF-α in response to fungi, a critical step required for the successful control of these pathogens.

The use of 4-acetoxy-2,2-dimethylbutanoyl protecting group for the C2-hydroxyl allows the selective formation of β-glycosides without producing α-glycosides. This very bulky protecting group can be removed under mild conditions.

(1,3;1,4)-β- D-glucan (β-glucan) is a plant cell wall hemicellulose and a main component of endosperm cell walls. The Cellulose Synthase F family of genes is involved in the synthesis of β-glucan. In this study full-length genomic sequences of CslF6 were obtained from multiple Avena species. Three unique alleles were found in each A. sativa line. Comparisons of these alleles to diploid Avena species allowed for identification of the genomic origin of each allele. The A and D genome alleles had identical amino acid sequences while the C-genome had 13 different amino acids. Global expression of CslF6 was completed at three developmental time point and three tissue types. RNAseq technology was utilized to determine genome specific expression patterns. Differential expression of genome specific-copies of CslF6 was found at all time points tested. Lower levels of C-genome expression of CslF6 were associated with increased levels of B-glucan.