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Nutritional supplement of lutein attenuates dopaminergic neurodegeneration induced by rotenone in rats
Abstracts of Pharma Nutrition 2013 / PharmaNutrition 2 (2014) 75–119
cumin, the predominant polyphenolic compound in the rhizome of Curcuma longa Linn., on suppression of PCSK9 expression in HepG2 cells. Methods and results: To investigate the effect of curcumin on PCSK9 gene expression, the levels of PCSK9 mRNA and proteins were determined by quantitative real-time PCR and Western blot analysis in curcumin-treated HepG2 cells. We found that curcumin significantly reduces intracellular mRNA and protein expression of PCSK9 in a dose-dependent manner. To investigate the mechanisms underlying the transcriptional inhibition of PCSK9 by curcumin, the curcumin-responsive sequence of PCSK9 promoter is determined by luciferase reporter assay. Our finding showed that the DNA region (−411 to −350) of PCSK9 promoter as a critical sequence for curcumin-mediated reduction of PCSK9 transcription. Within the curcumin-responsive region, a transcription factor hepatocyte nuclear factor 1␣ (HNF1␣) has been identified for activation of PCSK9 transcription. We further determined that curcumin decreases the levels of nuclear HNF1␣, but not SREBP2, in HepG2 cells. Finally, the statin-induced PCSK9 expression is also significantly suppressed by treatment of curcumin in HepG2 cells. Discussion: In conclusion, the above findings reveal that curcumin decreases PCSK9 expression through reduction of nuclear HNF1␣, which plays a vital role for promoting the transcription of PCSK9. This study thus enhances our understanding on curcumin may be beneficial for prevention and therapeutic use for familial hypercholesterolemia. Keywords: Curcumin; PCSK9; HNF1␣ http://dx.doi.org/10.1016/j.phanu.2013.11.055 [P15] Pim-1 kinase inhibitors induce radiosensitization in non-small cell lung cancer cells J. Kang ∗ , E. Kim, H. Seo, J.H. Lee, W. Kim, T. Kwon, B. Son, H.J. Yang, B. Youn Pusan National University, Republic of Korea Radiotherapy plays a critical role in the treatment of non-small cell lung cancer (NSCLC). However, radioresistance is a major barrier against increasing the efficiency of radiotherapy for NSCLC. To understand the mechanisms underlying NSCLC radioresistance, we previously focused on the potential involvement of PIM1, PRAS40, FOXO3a, 14-3-3, and protein phosphatases. Among these proteins, PIM1 functioned as an oncogene and was found to act as a crucial mediator in radioresistant NSCLC cells. Therefore, we investigated the use of PIM1-specific inhibitors as novel therapeutic drugs to regulate radiosensitivity in NSCLC. After structure-based drug selection, SGI-1776, ETP- 45299, and tryptanthrin were selected as candidates of PIM1 inhibitors that act as radiosensitizers. With irradiation, these drugs inhibited only PIM1 kinase activity without affecting PIM1 mRNA/protein levels or cellular localization. When PIM1 kinase activity was suppressed by these inhibitors, PRAS40 was not phosphorylated. Consequently, unphosphorylated PRAS40 did not form trimeric complexes with 14-3-3 and FOXO3a, leading to increased nuclear localization of FOXO3a. Nuclear FOXO3a promoted the expression of pro-apoptotic proteins such as Bim and FasL, resulting in a radiosensitizing effect on radioresistant NSCLC cells. Moreover, an in vivo xenograft mouse model confirmed this radiosensitizing effect induced by PIM1 inhibitors. In these model systems, tumor volume was significantly reduced by a combinational treatment with irradiation and PIM1 inhibitors compared to irradiation alone. Taken together, our findings provided evidence that PIM1-specific inhibitors, SGI-1776, ETP-45299, and tryptanthrin, can act as novel radiosensitizers to enhance the efficacy of
95
radiotherapy by inhibiting irradiation-induced signaling pathway associated with radioresistance. Keywords: Radiosensitizer; PIM1 inhibitors; Non-small cell lung cancer (NSCLC) http://dx.doi.org/10.1016/j.phanu.2013.11.056 [P16] Patenting in the European medical nutrition industry: Trends, opportunities and strategies T.C. Weenen 1,∗ , E. Pronker 2 , H.R. Commandeur 1 , E. Claassen 1,3 1
Erasmus University, The Netherlands Vacceleron, The Netherlands 3 Vrije Universiteit, The Netherlands 2
Medical nutrition products are specific nutritional compositions for intervention in disease progression and symptom alleviation. This industry finds itself on the interface between the food and pharmaceutical industry and even though it represents one of the fastest growing segments within the health and life sciences, it is still a relatively unknown industry. At present, insights concerning industry development and patenting in the European medical nutrition industry are limited. This research presents a systematic patent portfolio analysis of the industrial patenting trends and patenting strategy categorization of the 5 leading companies. Focusing on EU patent applications, we calculated company specific patent-, product- and market shares and average forward- and backward-citations. These indicators were combined to illustrate the European medical nutrition industry trends and company specific patent- and innovation-strategies. We found 222 European medical nutrition patent applications between 1990 up to 2010 with company specific patent shares ranging from 1 to 58%. The analysis of the industry trends shows that the industry currently resides in the growth phase and is estimated to reach the stage of maturation within two years with approximately 400 patents. Predominantly neurological diseases, cancer and diabetes show opportunity for future MN innovations while gastrointestinal and infection related diseases may have already reached a market saturation stage. Only three distinct patent strategies can be distinguished within this industry: the Prospector; the Analyzer; and the Reactor. Keywords: Innovation opportunities; Medical nutrition industry; Industry trends; Patent strategies http://dx.doi.org/10.1016/j.phanu.2013.11.057 [P17] Nutritional supplement of lutein attenuates dopaminergic neurodegeneration induced by rotenone in rats D.T. Makhija 1 , A.G. Jagtap 1,∗ , J.V. Shankaranarayanan 2 , S.K. Kulkarni 1 1 2
Deshpande 2 , J.
Bombay College of Pharmacy, India Omniactive Health Technologies, India
Introduction: Lutein, a well-known xanthophyll is present in green leafy vegetables like spinach and in some flowers like marigold. A well-established antioxidant, not synthesised in human body, but consumed through vegetables, and gets accumulated in the eye. Lutein is an established supplement for cataract and macular degeneration. In recent studies it has shown activity against some neurodegenerative disorders like Alzheimer’s disease. But its potential against Parkinson’s disease (PD) is yet to be explored.
96
Abstracts of Pharma Nutrition 2013 / PharmaNutrition 2 (2014) 75–119
Hence, potential of lutein against PD was evaluated using rotenone induced dopaminergic loss, which produces symptoms similar human PD. Aim: Assessment of anti-Parkinsonian potential of lutein using rotenone induced dopaminergic neurodegeneration in rats. Methodology: In vitro MTT assay was carried out on synaptosomal fractions of rat brain using rotenone as apoptotic agent. Lutein at concentrations of 2, 5, 10 and 20 g/ml was studied. In in vivo studies, lutein was evaluated at 50, 100 mg/kg (p.o.), using levodopa and carbidopa as standard. Lutein was administered 1 h prior to rotenone (2 mg/kg, sc). This dosing was continued for 21 days. Lutein was combined with various doses of levodopa and carbidopa to assess its potential in combination. Various behavioural and biochemical parameters were evaluated at various time points. Studies in TH immunoreactivity are in progress. Results: Lutein at concentration of 10 g/ml exhibited significant anti-apoptotic activity against rotenone in MTT assay. In, in vivo studies, it was observed that diseased control group was significantly (P < 0.05) different from normal control. Lutein at both the doses significantly (P < 0.05) attenuated all behavioural alterations and increased the brain levels of dopamine, NE, 5HT. There was no significant difference between lutein 50 and 100 mg/kg. Hence, combination studies were done using lutein 50 mg/kg. Amongst various combinations tried, there was no significant difference in activity between the group which received (levodopa 75 mg/kg + carbidopa 25 mg/kg) and (lutein 50 mg/kg + levodopa 25 mg/kg + carbidopa 6.25 mg/kg) treated group. Hence dose of levodopa can be reduced with lutein supplementation. Conclusion: Thus, it can be concluded that lutein present in a healthy diet can reduce the risk of PD and other neurodegenerative disorders onslaught. Further, its supplementation with conventional levodopa therapy can help in reducing the dose of levodopa and its side effects. Keywords: Lutein; Parkinson’s disease; Rotenone; Nutritional supplement http://dx.doi.org/10.1016/j.phanu.2013.11.058 [P18] Improvement by Eviprostat treatment of bladder dysfunction and altered levels of pharmacological receptors and urinary cytokines in rats with cyclophosphamide-induced cystitis S. Yamada ∗ , S. Nasrin, E. Masuda, Y. Ito University of Shizuoka, Japan Interstitial cystitis (IC) is a chronic, abacterial inflammatory disease of bladder characterized by urinary frequency, urgency and suprapubic pain associated with bladder filling and relieved by voiding, but its exact etiology and pathogenesis remain unclear and effective treatment is not established. The present study characterized pharmacological effects of a phytotherapeutic agent, Eviprostat (EVI) in rats with cystitis induced by cyclophosphamide (CYP). Cystitis model was induced by injecting CYP in female SpragueDawley rats. Following repeated oral administration of EVI for 7 days in CYP-treated rats, urinary function was monitored. Muscarinic and purinergic receptors in rat tissues were measured by radioreceptor assays using (N-methyl-3 H) scopolamine methyl chloride ([3 H]NMS) and ␣-methylene-ATP (2,8-3 H) tetrasodium salt ([3 H]␣-MeATP), respectively, and binding parameters of apparent dissociation constant (Kd ) and maximal number of binding sites (Bmax ) were estimated by nonlinear regression analysis. Urinary cytokines were measured by ELISA kits.
In the cystometry of CYP-treated rats compared with sham rats, micturition interval and micturition volume were significantly decreased and the frequency of micturition, basal pressure, and residual urine volume were significantly increased. Repeated administration of EVI significantly increased the micturition interval and micturition volume and decreased the frequency of micturition, basal pressure, and residual urine volume compared with CYP-treated rats. Bmax for specific binding of [3 H]NMS and [3 H]␣-MeATP was significantly decreased in the bladder of CYPtreated rats compared with sham rats. There was significant increase in Bmax for [3 H]NMS and [3 H]␣-MeATP in the bladder of rats treated with CYP+EVI, compared with CYP-treated rat bladder. The elevation in urinary cytokine level in CYP-treated rats was effectively attenuated by repeated treatment with EVI. The present results revealed that alteration of urodynamic parameters, pharmacologically relevant receptors and urinary cytokines in CYP-treated rats was attenuated by the repeated treatment with EVI, suggesting pharmacological usefulness of EVI in the therapy of cystitis. Keywords: Cystitis; Plant extract; Urination http://dx.doi.org/10.1016/j.phanu.2013.11.059 [P19] Protection of cow’s milk allergy (CMA) by non digestible oligosaccharides in mice is associated with altered mucosal immune polarization and abrogated by IL10 and TGFbeta neutralization J. Kerperien 1 , S. de Kivit 1 , B.C.A.M. van Esch 1,2 , G.A. Hofman 1 , L. Boon 3 , L.M.J. Knippels 1,2,∗ , J. Garssen 1,2 , L.E.M. Willemsen 1 1
Utrecht University, The Netherlands Danone Research, The Netherlands 3 Bioceros BV, The Netherlands 2
We determined the effects of dietary intervention with shortchain galacto-(scGOS), longchain fructo-(lcFOS) and/or pectin derived acidic-(pAOS) oligosaccharides on the mucosal immune response and assess the contribution of IL10 and TGFbeta in the protective effect of the diet in CMA. Mice were fed control or 1% scGOS/lcFOS/pAOS (9:1:2) diet prior to and during oral whey sensitization (5*, once a week). In a second study mice were injected 24 h prior to each sensitization with anti-IL10R, anti-TGF or isotype antibodies. The acute allergic skin response, anaphylaxis, immunoglobulin’s and mMCP1, intestinal cytokine and transcription factor expression (qPCR/IHC) were determined and MLN were characterized. scGOS/lcFOS/pAOS reduced the acute allergic skin response over 50% and abrogated anaphylaxis compared to whey sensitized mice fed control diet. In the middle ileum of whey sensitized mice mRNA expression of Treg, TGFbeta, IL10 and Th2 type IL-4 and Gata3 was enhanced compared to sham sensitized mice (qPCR/IHC). Furthermore, scGOS/lcFOS/pAOS enhanced the expression of IFNgamma, Tbet, and IL17 mRNA compared to control diet in sensitized mice (p < 0.05). The reduction of the allergic skin response in CMA mice fed scGOS/lcFOS/pAOS was reversed by both IL10 and TGFbeta neutralizing antibodies. Dietary scGOS/lcFOS/pAOS can probably enhance intestinal Treg, Th1, Th2 and Th17 type immune activation in association with reduced allergic effector response in CMA, important will be the balance. IL10 and TGFbeta may be of key importance for the protective effect of the diet. Because of these promising results (qPCR/IHC) functionally will be assessed in the near future using intestinal lamina propria isolation (FACS).
cumin, the predominant polyphenolic compound in the rhizome of Curcuma longa Linn., on suppression of PCSK9 expression in HepG2 cells. Methods and results: To investigate the effect of curcumin on PCSK9 gene expression, the levels of PCSK9 mRNA and proteins were determined by quantitative real-time PCR and Western blot analysis in curcumin-treated HepG2 cells. We found that curcumin significantly reduces intracellular mRNA and protein expression of PCSK9 in a dose-dependent manner. To investigate the mechanisms underlying the transcriptional inhibition of PCSK9 by curcumin, the curcumin-responsive sequence of PCSK9 promoter is determined by luciferase reporter assay. Our finding showed that the DNA region (−411 to −350) of PCSK9 promoter as a critical sequence for curcumin-mediated reduction of PCSK9 transcription. Within the curcumin-responsive region, a transcription factor hepatocyte nuclear factor 1␣ (HNF1␣) has been identified for activation of PCSK9 transcription. We further determined that curcumin decreases the levels of nuclear HNF1␣, but not SREBP2, in HepG2 cells. Finally, the statin-induced PCSK9 expression is also significantly suppressed by treatment of curcumin in HepG2 cells. Discussion: In conclusion, the above findings reveal that curcumin decreases PCSK9 expression through reduction of nuclear HNF1␣, which plays a vital role for promoting the transcription of PCSK9. This study thus enhances our understanding on curcumin may be beneficial for prevention and therapeutic use for familial hypercholesterolemia. Keywords: Curcumin; PCSK9; HNF1␣ http://dx.doi.org/10.1016/j.phanu.2013.11.055 [P15] Pim-1 kinase inhibitors induce radiosensitization in non-small cell lung cancer cells J. Kang ∗ , E. Kim, H. Seo, J.H. Lee, W. Kim, T. Kwon, B. Son, H.J. Yang, B. Youn Pusan National University, Republic of Korea Radiotherapy plays a critical role in the treatment of non-small cell lung cancer (NSCLC). However, radioresistance is a major barrier against increasing the efficiency of radiotherapy for NSCLC. To understand the mechanisms underlying NSCLC radioresistance, we previously focused on the potential involvement of PIM1, PRAS40, FOXO3a, 14-3-3, and protein phosphatases. Among these proteins, PIM1 functioned as an oncogene and was found to act as a crucial mediator in radioresistant NSCLC cells. Therefore, we investigated the use of PIM1-specific inhibitors as novel therapeutic drugs to regulate radiosensitivity in NSCLC. After structure-based drug selection, SGI-1776, ETP- 45299, and tryptanthrin were selected as candidates of PIM1 inhibitors that act as radiosensitizers. With irradiation, these drugs inhibited only PIM1 kinase activity without affecting PIM1 mRNA/protein levels or cellular localization. When PIM1 kinase activity was suppressed by these inhibitors, PRAS40 was not phosphorylated. Consequently, unphosphorylated PRAS40 did not form trimeric complexes with 14-3-3 and FOXO3a, leading to increased nuclear localization of FOXO3a. Nuclear FOXO3a promoted the expression of pro-apoptotic proteins such as Bim and FasL, resulting in a radiosensitizing effect on radioresistant NSCLC cells. Moreover, an in vivo xenograft mouse model confirmed this radiosensitizing effect induced by PIM1 inhibitors. In these model systems, tumor volume was significantly reduced by a combinational treatment with irradiation and PIM1 inhibitors compared to irradiation alone. Taken together, our findings provided evidence that PIM1-specific inhibitors, SGI-1776, ETP-45299, and tryptanthrin, can act as novel radiosensitizers to enhance the efficacy of
95
radiotherapy by inhibiting irradiation-induced signaling pathway associated with radioresistance. Keywords: Radiosensitizer; PIM1 inhibitors; Non-small cell lung cancer (NSCLC) http://dx.doi.org/10.1016/j.phanu.2013.11.056 [P16] Patenting in the European medical nutrition industry: Trends, opportunities and strategies T.C. Weenen 1,∗ , E. Pronker 2 , H.R. Commandeur 1 , E. Claassen 1,3 1
Erasmus University, The Netherlands Vacceleron, The Netherlands 3 Vrije Universiteit, The Netherlands 2
Medical nutrition products are specific nutritional compositions for intervention in disease progression and symptom alleviation. This industry finds itself on the interface between the food and pharmaceutical industry and even though it represents one of the fastest growing segments within the health and life sciences, it is still a relatively unknown industry. At present, insights concerning industry development and patenting in the European medical nutrition industry are limited. This research presents a systematic patent portfolio analysis of the industrial patenting trends and patenting strategy categorization of the 5 leading companies. Focusing on EU patent applications, we calculated company specific patent-, product- and market shares and average forward- and backward-citations. These indicators were combined to illustrate the European medical nutrition industry trends and company specific patent- and innovation-strategies. We found 222 European medical nutrition patent applications between 1990 up to 2010 with company specific patent shares ranging from 1 to 58%. The analysis of the industry trends shows that the industry currently resides in the growth phase and is estimated to reach the stage of maturation within two years with approximately 400 patents. Predominantly neurological diseases, cancer and diabetes show opportunity for future MN innovations while gastrointestinal and infection related diseases may have already reached a market saturation stage. Only three distinct patent strategies can be distinguished within this industry: the Prospector; the Analyzer; and the Reactor. Keywords: Innovation opportunities; Medical nutrition industry; Industry trends; Patent strategies http://dx.doi.org/10.1016/j.phanu.2013.11.057 [P17] Nutritional supplement of lutein attenuates dopaminergic neurodegeneration induced by rotenone in rats D.T. Makhija 1 , A.G. Jagtap 1,∗ , J.V. Shankaranarayanan 2 , S.K. Kulkarni 1 1 2
Deshpande 2 , J.
Bombay College of Pharmacy, India Omniactive Health Technologies, India
Introduction: Lutein, a well-known xanthophyll is present in green leafy vegetables like spinach and in some flowers like marigold. A well-established antioxidant, not synthesised in human body, but consumed through vegetables, and gets accumulated in the eye. Lutein is an established supplement for cataract and macular degeneration. In recent studies it has shown activity against some neurodegenerative disorders like Alzheimer’s disease. But its potential against Parkinson’s disease (PD) is yet to be explored.
96
Abstracts of Pharma Nutrition 2013 / PharmaNutrition 2 (2014) 75–119
Hence, potential of lutein against PD was evaluated using rotenone induced dopaminergic loss, which produces symptoms similar human PD. Aim: Assessment of anti-Parkinsonian potential of lutein using rotenone induced dopaminergic neurodegeneration in rats. Methodology: In vitro MTT assay was carried out on synaptosomal fractions of rat brain using rotenone as apoptotic agent. Lutein at concentrations of 2, 5, 10 and 20 g/ml was studied. In in vivo studies, lutein was evaluated at 50, 100 mg/kg (p.o.), using levodopa and carbidopa as standard. Lutein was administered 1 h prior to rotenone (2 mg/kg, sc). This dosing was continued for 21 days. Lutein was combined with various doses of levodopa and carbidopa to assess its potential in combination. Various behavioural and biochemical parameters were evaluated at various time points. Studies in TH immunoreactivity are in progress. Results: Lutein at concentration of 10 g/ml exhibited significant anti-apoptotic activity against rotenone in MTT assay. In, in vivo studies, it was observed that diseased control group was significantly (P < 0.05) different from normal control. Lutein at both the doses significantly (P < 0.05) attenuated all behavioural alterations and increased the brain levels of dopamine, NE, 5HT. There was no significant difference between lutein 50 and 100 mg/kg. Hence, combination studies were done using lutein 50 mg/kg. Amongst various combinations tried, there was no significant difference in activity between the group which received (levodopa 75 mg/kg + carbidopa 25 mg/kg) and (lutein 50 mg/kg + levodopa 25 mg/kg + carbidopa 6.25 mg/kg) treated group. Hence dose of levodopa can be reduced with lutein supplementation. Conclusion: Thus, it can be concluded that lutein present in a healthy diet can reduce the risk of PD and other neurodegenerative disorders onslaught. Further, its supplementation with conventional levodopa therapy can help in reducing the dose of levodopa and its side effects. Keywords: Lutein; Parkinson’s disease; Rotenone; Nutritional supplement http://dx.doi.org/10.1016/j.phanu.2013.11.058 [P18] Improvement by Eviprostat treatment of bladder dysfunction and altered levels of pharmacological receptors and urinary cytokines in rats with cyclophosphamide-induced cystitis S. Yamada ∗ , S. Nasrin, E. Masuda, Y. Ito University of Shizuoka, Japan Interstitial cystitis (IC) is a chronic, abacterial inflammatory disease of bladder characterized by urinary frequency, urgency and suprapubic pain associated with bladder filling and relieved by voiding, but its exact etiology and pathogenesis remain unclear and effective treatment is not established. The present study characterized pharmacological effects of a phytotherapeutic agent, Eviprostat (EVI) in rats with cystitis induced by cyclophosphamide (CYP). Cystitis model was induced by injecting CYP in female SpragueDawley rats. Following repeated oral administration of EVI for 7 days in CYP-treated rats, urinary function was monitored. Muscarinic and purinergic receptors in rat tissues were measured by radioreceptor assays using (N-methyl-3 H) scopolamine methyl chloride ([3 H]NMS) and ␣-methylene-ATP (2,8-3 H) tetrasodium salt ([3 H]␣-MeATP), respectively, and binding parameters of apparent dissociation constant (Kd ) and maximal number of binding sites (Bmax ) were estimated by nonlinear regression analysis. Urinary cytokines were measured by ELISA kits.
In the cystometry of CYP-treated rats compared with sham rats, micturition interval and micturition volume were significantly decreased and the frequency of micturition, basal pressure, and residual urine volume were significantly increased. Repeated administration of EVI significantly increased the micturition interval and micturition volume and decreased the frequency of micturition, basal pressure, and residual urine volume compared with CYP-treated rats. Bmax for specific binding of [3 H]NMS and [3 H]␣-MeATP was significantly decreased in the bladder of CYPtreated rats compared with sham rats. There was significant increase in Bmax for [3 H]NMS and [3 H]␣-MeATP in the bladder of rats treated with CYP+EVI, compared with CYP-treated rat bladder. The elevation in urinary cytokine level in CYP-treated rats was effectively attenuated by repeated treatment with EVI. The present results revealed that alteration of urodynamic parameters, pharmacologically relevant receptors and urinary cytokines in CYP-treated rats was attenuated by the repeated treatment with EVI, suggesting pharmacological usefulness of EVI in the therapy of cystitis. Keywords: Cystitis; Plant extract; Urination http://dx.doi.org/10.1016/j.phanu.2013.11.059 [P19] Protection of cow’s milk allergy (CMA) by non digestible oligosaccharides in mice is associated with altered mucosal immune polarization and abrogated by IL10 and TGFbeta neutralization J. Kerperien 1 , S. de Kivit 1 , B.C.A.M. van Esch 1,2 , G.A. Hofman 1 , L. Boon 3 , L.M.J. Knippels 1,2,∗ , J. Garssen 1,2 , L.E.M. Willemsen 1 1
Utrecht University, The Netherlands Danone Research, The Netherlands 3 Bioceros BV, The Netherlands 2
We determined the effects of dietary intervention with shortchain galacto-(scGOS), longchain fructo-(lcFOS) and/or pectin derived acidic-(pAOS) oligosaccharides on the mucosal immune response and assess the contribution of IL10 and TGFbeta in the protective effect of the diet in CMA. Mice were fed control or 1% scGOS/lcFOS/pAOS (9:1:2) diet prior to and during oral whey sensitization (5*, once a week). In a second study mice were injected 24 h prior to each sensitization with anti-IL10R, anti-TGF or isotype antibodies. The acute allergic skin response, anaphylaxis, immunoglobulin’s and mMCP1, intestinal cytokine and transcription factor expression (qPCR/IHC) were determined and MLN were characterized. scGOS/lcFOS/pAOS reduced the acute allergic skin response over 50% and abrogated anaphylaxis compared to whey sensitized mice fed control diet. In the middle ileum of whey sensitized mice mRNA expression of Treg, TGFbeta, IL10 and Th2 type IL-4 and Gata3 was enhanced compared to sham sensitized mice (qPCR/IHC). Furthermore, scGOS/lcFOS/pAOS enhanced the expression of IFNgamma, Tbet, and IL17 mRNA compared to control diet in sensitized mice (p < 0.05). The reduction of the allergic skin response in CMA mice fed scGOS/lcFOS/pAOS was reversed by both IL10 and TGFbeta neutralizing antibodies. Dietary scGOS/lcFOS/pAOS can probably enhance intestinal Treg, Th1, Th2 and Th17 type immune activation in association with reduced allergic effector response in CMA, important will be the balance. IL10 and TGFbeta may be of key importance for the protective effect of the diet. Because of these promising results (qPCR/IHC) functionally will be assessed in the near future using intestinal lamina propria isolation (FACS).