- Email: [email protected]
Microbial metabolites produced by the colonic microbiota from prebiotics as drivers for immunomodulation in the host
118
Abstracts of Pharma Nutrition 2013 / PharmaNutrition 2 (2014) 75–119
tion to inflammatory effects, food allergy an metabolic syndrome. The first tier of the strategy includes assays to assess whether a natural products, i.e. their chemical contents, can be metabolically converted or whether it has oxidant and/or mutagenic potential. The second tier of the strategy focuses on organ-specific assays, in particular intestinal and adipocyte models. All of the assays of the proposed strategy require extensive standardization and validation for screening and selection purposes, but also to meet regulatory requirements. A series of assays has been explored to detect oxidant potential, including the “Antioxidant Protection in Erythrocytes” (CAP-e) assay and a Nrf2-Luc reporter cell assay. Of these two assays, the Cap-e test, tested with 7 antioxidants appears most suitable as high throughput test. To explore immunomodulating as well as gut permeability effects of compounds, including preand probiotics, we use the transwell CACO2-system. The co-culture model of adipocytes (differentiated murine 3T3) and macrophages (RAW246.7) is set up to assess inflammatory effects of compounds, based on production of adiponectine and IL-6. Data of experiments with selected plant and food-derived substances will be presented. Experiments with additional compounds will allow further assessment of the applicability domain of the various assays. This tiered approach aims to facilitate strategic decision making in the R&D process of new natural products. Keywords: Non animal test strategy; Food and plant substances; Gut health; Inflammation http://dx.doi.org/10.1016/j.phanu.2013.11.123 [OP4] A prebiotic multi-fiber mix inhibits inflammation in DSS induced colitis and leads to a relative increase in MLN tregs A. Hartog 1,2 , J. Bastiaans 1,2 , T. Wehkamp 1,2 , F.N. Belle 2,3 , P. de Graaff 2 , J. Garssen 1,2 , L.F. Harthoorn 2,3 , A.P. Vos 1,2,∗ 1
Utrecht University, The Netherlands Danone Research, The Netherlands 3 Nutricia Advanced Medical Nutrition, The Netherlands 2
Inflammatory bowel diseases (IBDs) are inflammatory disorders caused by multiple factors. The dextran sodium sulfate (DSS)-induced colitis mouse model is widely used to study IBDrelated mechanisms and interventions. Literature indicates that an imbalance between T helper (Th) subsets contributes to the DSS-induced inflammation. The present study aimed to analyze the effect of a prebiotic multi-fiber mix (MF), comprising betagalacto-oligosaccharide, fructan, non-digestible alpha-glucan, and hemicellulose, on DSS-induced colitis. Oral administration of MF reduced the DSS-induced weight loss and changes in stool consistency. Moreover, MF intake counteracted the increase in serum amyloid A and the mesenteric lymph node (MLN) cell number. After 7 days of DSS treatment the number of Th1, Th2 and Treg cells in the MLN, as analyzed by flow cytometry, was increased significantly to 268%, 357% and 327% of control levels, respectively. The number of Th17 cells increased to 680%, pointing to a strong Th17driven disease process in the MLN. MF intake counteracted the DSS induced increase in T cell numbers in the MLN. The Tregs remained more abundant in the MLN, resulting in a relative increase in Treg cell numbers compared to the other T cell subsets. Concluding, DSS induced colitis leads to strong Th17-driven responses in the MLN. Oral intake of MF reduced local and systemic disease symptoms. The reduction in inflammation might be mediated, at least for a part, by the relative increase of Treg cells in the MLN. Keywords: IBD; Treg; Multi-fiber; DSS http://dx.doi.org/10.1016/j.phanu.2013.11.124
[OP5] Microbial metabolites produced by the colonic microbiota from prebiotics as drivers for immunomodulation in the host K. Venema 1,2 1 2
Top Institute Food & Nutrition, The Netherlands TNO Healthy Living, The Netherlands
The colonic microbiota ferments dietary substrates that are not digested in the upper gastrointestinal tract. The most important dietary substrates that make it to the colon are prebiotics and other complex carbohydrates. Fermentation of these prebiotics and complex carbohydrates leads to the production of so-called short-chain fatty acids (SCFA). These SCFA (primarily acetate, propionate and butyrate) have been shown to have several beneficial effects on the host. Apart from functioning as energy substrate (e.g., butyrate is the major energy source for colonocytes), these molecules have also been shown to function as immune signaling molecules. For instance, butyrate has been shown to reduce inflammation in inflammatory bowel disease, possibly through its histone deacetylase inhibitory activity. Propionate has effects on cells in the adipose tissue. The SCFA molecules are hypothesized to function through the G-protein coupled receptors GPCR41 and/or GPCR43. However, we have shown that additional pathways must be active in adipose tissue, as signaling of propionate was not completely blocked by the GPCR inhibitor pertusis toxin. Several tools were combined to study SCFA production by the colonic microbiota and their potential health effects. These tools include a sophisticated, validated in vitro model of the colon including a complex microbiota, 13 C stable isotope technology, and high through-put reporter cell-lines. The microbiota used originates from several different volunteers, e.g., lean or obese, different enterotypes, different ages, healthy or diseased. Using the combined set of tools we have shown that different microbiotas lead to production of different ratios of SCFA (and other microbial metabolites), and thus may be portrayed to have different health effects in the different individuals whom the microbiotas stem from. Using specific prebiotics, the activity of the microbiota can be directed towards production of certain SCFA, and through this the immunomodulation (or ‘immune assistance’) in these individuals can be influenced. Keywords: Immune assistance; Prebiotics; SCFA; Colonic microbiota http://dx.doi.org/10.1016/j.phanu.2013.11.125 [OP3] Evaluation of the cytotoxicity and antiviral activity of zingiber officinale against new castle disease virus A. Javeed ∗ , A. Iqbal, M. Ashraf, I. Altaf University of Veterinary and Animal Science, Lahore, Pakistan Introduction: Ethnomedicines are being used in the control of poultry diseases in many parts of the world. The present study was designed to investigate the antiviral property of the water extract of Zingiber officinale against Newcastle disease virus. Methods: Ninty specific antibody negative embryonated chicken eggs and a field strain of Newcastle disease virus were used to test for the antiviral activity of the water extract of Z. officinale. Following a 2 h exposure of the virus to eight graded concentrations of the extract, it was incubated at 37 ◦ C and observed 12-hourly for mortality. Dead embryos were spot tested for haemagglutinating activity. The 4HA concentration of the virus and normal saline alone were inoculated as positive and negative controls, respectively.
Abstracts of Pharma Nutrition 2013 / PharmaNutrition 2 (2014) 75–119
tion to inflammatory effects, food allergy an metabolic syndrome. The first tier of the strategy includes assays to assess whether a natural products, i.e. their chemical contents, can be metabolically converted or whether it has oxidant and/or mutagenic potential. The second tier of the strategy focuses on organ-specific assays, in particular intestinal and adipocyte models. All of the assays of the proposed strategy require extensive standardization and validation for screening and selection purposes, but also to meet regulatory requirements. A series of assays has been explored to detect oxidant potential, including the “Antioxidant Protection in Erythrocytes” (CAP-e) assay and a Nrf2-Luc reporter cell assay. Of these two assays, the Cap-e test, tested with 7 antioxidants appears most suitable as high throughput test. To explore immunomodulating as well as gut permeability effects of compounds, including preand probiotics, we use the transwell CACO2-system. The co-culture model of adipocytes (differentiated murine 3T3) and macrophages (RAW246.7) is set up to assess inflammatory effects of compounds, based on production of adiponectine and IL-6. Data of experiments with selected plant and food-derived substances will be presented. Experiments with additional compounds will allow further assessment of the applicability domain of the various assays. This tiered approach aims to facilitate strategic decision making in the R&D process of new natural products. Keywords: Non animal test strategy; Food and plant substances; Gut health; Inflammation http://dx.doi.org/10.1016/j.phanu.2013.11.123 [OP4] A prebiotic multi-fiber mix inhibits inflammation in DSS induced colitis and leads to a relative increase in MLN tregs A. Hartog 1,2 , J. Bastiaans 1,2 , T. Wehkamp 1,2 , F.N. Belle 2,3 , P. de Graaff 2 , J. Garssen 1,2 , L.F. Harthoorn 2,3 , A.P. Vos 1,2,∗ 1
Utrecht University, The Netherlands Danone Research, The Netherlands 3 Nutricia Advanced Medical Nutrition, The Netherlands 2
Inflammatory bowel diseases (IBDs) are inflammatory disorders caused by multiple factors. The dextran sodium sulfate (DSS)-induced colitis mouse model is widely used to study IBDrelated mechanisms and interventions. Literature indicates that an imbalance between T helper (Th) subsets contributes to the DSS-induced inflammation. The present study aimed to analyze the effect of a prebiotic multi-fiber mix (MF), comprising betagalacto-oligosaccharide, fructan, non-digestible alpha-glucan, and hemicellulose, on DSS-induced colitis. Oral administration of MF reduced the DSS-induced weight loss and changes in stool consistency. Moreover, MF intake counteracted the increase in serum amyloid A and the mesenteric lymph node (MLN) cell number. After 7 days of DSS treatment the number of Th1, Th2 and Treg cells in the MLN, as analyzed by flow cytometry, was increased significantly to 268%, 357% and 327% of control levels, respectively. The number of Th17 cells increased to 680%, pointing to a strong Th17driven disease process in the MLN. MF intake counteracted the DSS induced increase in T cell numbers in the MLN. The Tregs remained more abundant in the MLN, resulting in a relative increase in Treg cell numbers compared to the other T cell subsets. Concluding, DSS induced colitis leads to strong Th17-driven responses in the MLN. Oral intake of MF reduced local and systemic disease symptoms. The reduction in inflammation might be mediated, at least for a part, by the relative increase of Treg cells in the MLN. Keywords: IBD; Treg; Multi-fiber; DSS http://dx.doi.org/10.1016/j.phanu.2013.11.124
[OP5] Microbial metabolites produced by the colonic microbiota from prebiotics as drivers for immunomodulation in the host K. Venema 1,2 1 2
Top Institute Food & Nutrition, The Netherlands TNO Healthy Living, The Netherlands
The colonic microbiota ferments dietary substrates that are not digested in the upper gastrointestinal tract. The most important dietary substrates that make it to the colon are prebiotics and other complex carbohydrates. Fermentation of these prebiotics and complex carbohydrates leads to the production of so-called short-chain fatty acids (SCFA). These SCFA (primarily acetate, propionate and butyrate) have been shown to have several beneficial effects on the host. Apart from functioning as energy substrate (e.g., butyrate is the major energy source for colonocytes), these molecules have also been shown to function as immune signaling molecules. For instance, butyrate has been shown to reduce inflammation in inflammatory bowel disease, possibly through its histone deacetylase inhibitory activity. Propionate has effects on cells in the adipose tissue. The SCFA molecules are hypothesized to function through the G-protein coupled receptors GPCR41 and/or GPCR43. However, we have shown that additional pathways must be active in adipose tissue, as signaling of propionate was not completely blocked by the GPCR inhibitor pertusis toxin. Several tools were combined to study SCFA production by the colonic microbiota and their potential health effects. These tools include a sophisticated, validated in vitro model of the colon including a complex microbiota, 13 C stable isotope technology, and high through-put reporter cell-lines. The microbiota used originates from several different volunteers, e.g., lean or obese, different enterotypes, different ages, healthy or diseased. Using the combined set of tools we have shown that different microbiotas lead to production of different ratios of SCFA (and other microbial metabolites), and thus may be portrayed to have different health effects in the different individuals whom the microbiotas stem from. Using specific prebiotics, the activity of the microbiota can be directed towards production of certain SCFA, and through this the immunomodulation (or ‘immune assistance’) in these individuals can be influenced. Keywords: Immune assistance; Prebiotics; SCFA; Colonic microbiota http://dx.doi.org/10.1016/j.phanu.2013.11.125 [OP3] Evaluation of the cytotoxicity and antiviral activity of zingiber officinale against new castle disease virus A. Javeed ∗ , A. Iqbal, M. Ashraf, I. Altaf University of Veterinary and Animal Science, Lahore, Pakistan Introduction: Ethnomedicines are being used in the control of poultry diseases in many parts of the world. The present study was designed to investigate the antiviral property of the water extract of Zingiber officinale against Newcastle disease virus. Methods: Ninty specific antibody negative embryonated chicken eggs and a field strain of Newcastle disease virus were used to test for the antiviral activity of the water extract of Z. officinale. Following a 2 h exposure of the virus to eight graded concentrations of the extract, it was incubated at 37 ◦ C and observed 12-hourly for mortality. Dead embryos were spot tested for haemagglutinating activity. The 4HA concentration of the virus and normal saline alone were inoculated as positive and negative controls, respectively.