OF THE HEALTH BENEFITS OF FOOD INGREDIENTS TO SUPPORT RESISTANCE - PowerPoint PPT Presentation

TAKING UP THE CHALLENGE FOR SUBSTANTIATION OF THE HEALTH BENEFITS OF FOOD INGREDIENTS TO SUPPORT RESISTANCE AGAINST INFECTION Anita Hartog, Senior Scientist Nutrition & Health Anita.Hartog@nizo.com 11-12-2019

CONFLICT OF INTEREST Potentially relevant company or other financial relationships: • NIZO is a Contract Research Organisation for food and health • We collaborate with …

NIZO - FOR BETTER FOOD & HEALTH Application & Research Center • Independent, privately owned Contract Research Processing Centre Organisation for food and health • HQ in Ede, The Netherlands (Food Valley) • Founded in 1948; 100 professionals

TRACK RECORD 9 of the global top 10 6 of the global top 10 100 professionals The global top 5 Dairy Companies Ingredient companies Infant & Clinical at NIZO companies Number of publications per year 60 on average 6 of the global top 10 6 of the global top 10 17 General labs 16 BSL-I labs* Consumer goods companies Personal care companies 4 BSL-II labs* Number of consortia 5 Food grade labs on average 5-10 per year *also for GMOs

Nutrition & Health

INFECTIOUS DISEASES: AMONG THE TOP 10 GLOBAL CAUSES OF DEATH

THE YOUNG AND THE OLD: IMMUNE COMPROMISED Immune immaturity Immunosenescence Simon AK et al . Proc. R. Soc. B 2015;282: 20143085.

NUTRITION CAN SUPPORT RESISTANCE AGAINST INFECTION • Potential targets for nutritional support of resistance against infection: • General health status: • Adequate nutritional status • Immune system: Dietary Plant fibers ingredients • General immune response • Immune response induced by infection • Direct anti-pathogenic: Milk Vitamins components • Reduced adhesion/invasion of pathogens • Anti-microbial activity • Microbiota community: • Adequate diversity • Adequate composition

FOOD INGREDIENTS WITH APPROVED HEALTH CLAIM Health claims on foods are aimed at maintenance and improvement of Immune function : Absorption of health. Vitamins: A, B6, micronutrients: B9, B12, C, D Vitamins: Minerals: C (Fe absorption) Zn, Cu, Fe, Se EFSA claims on the immune system, the gastrointestinal tract, and defence against pathogens (EFSA NDA panel*): Bowel function/ defecation : Chicory inulin, dried plums, rye fibre, lactitol, wheat and oat and barley bran fibre * EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA). EFSA Journal 2016;14(1):4369.

IDENTIFICATION OF NEW FOOD INGREDIENTS WITH A POTENTIAL HEALTH BENEFIT RELATED TO INFECTION RESISTANCE • Literature : • Ingredients with known mechanisms of action • Use smart in silico tools ; text mining to identify new candidates • In vitro models : medium-high throughput screening • Human cell-lines: ✓ Gut/skin/lung epithelial cells: barrier function, pathogen adhesion, modulation of inflammation ✓ Immune cells: support of immune function, modulation of systemic inflammation • Micro-models of microbial communities ✓ Support of healthy gut microbiota composition and function

PROBLEM How to translate in vitro effects to a health benefit in healthy subjects? • Gap between in vitro /animal models and field studies creates high attrition rate High Attrition Rate Field trial Human Challenge Model in vitro assays Human Challenge Model (claim support in Animal models? (Proof of Concept in Human) (HT, HC, mechanism) (Proof of Concept in Human) target population) Data Driven Decision • Human Challenge Models (HCM) as Proof of Concept (PoC) to enhance the efficacy of health research

FUNCTION CLAIMS FOR DEFENSE AGAINST PATHOGENS Appropriate outcome measures • Number of episodes and severity or duration of infection. Establish infectious nature of the disease: 1. By clinical diagnosis 2. By use of validated questionnaires for self-reported data 3. By microbiological data depending on the type of the infection • Reduction of the presence of specific pathogens, their toxins or other virulence factors. Justify relevance: 1. By the magnitude of reduction or 2. By evidence of a reduction in clinical outcomes related to infection accompanying the reduction in pathogens/toxins * EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA). EFSA Journal 2016;14(1):4369.

WHAT IS HEALTH? • Health has always been regarded as the absence of disease . • Health can also be defined as: the body’s ability to adapt to changing circumstances and to show sufficient resilience under conditions of social, physical and emotional disturbance ( challenge ) to which we are exposed from time to time. Huber M. et al , BMJ. 2011;343:d4163

HOW TO DEMONSTRATE A HEALTH BENEFIT IN HEALTHY SUBJECTS? • Human Challenge Models (HCM): based on “ health is the ability to adapt ” • Challenge = applying a stressor to healthy people and measure resilience to this stress • Examples: pathogens, vaccination, high calorie meal, prescription drugs Both apparently healthy Stressor / Challenge Subject 1 Subject 2 Subject 2 Subject 1 Dietary intervention Dietary intervention Hardly any change Measurable change

WHY CHALLENGE MODELS? • Healthy subjects - Health claims on foods are aimed at maintenance and improvement of health • Effect size - Stress resilience is a more sensitive marker of health than steady state markers • Controlled stress exposure • Limited number of study subjects • Limited study duration

NIZO CHALLENGE MODELS FOR INFECTION RESISTANCE • Diarrheagenic E. coli challenge* • Challenge: E. coli E1392/75-2A • Outcomes: Gastro-intestinal symptoms, Bristol Stool Score, stool frequency, % of fecal wet weight • Vaccination** • Challenge: Oral cholera vaccine, parenteral hepatitis B vaccine • Outcomes: Vaccine-specific IgA, IgG, % responders • Rhinovirus challenge*** • Challenge: HRV-16 • Outcomes: Common cold symptoms & viral load *Bovee-Oudenhoven et al. Gastroenterology 2003;125(2):469-76., *Ten Bruggencate et al. J Nutr. 2016;146(2):249-55., **Van Splunter et al. Mucosal Immunol 2018;11(4):1254-64., ***Koch et al. PLoS One 2018;13(2):e0191937. & ***Turner et al. Benef Microbes. 2017;8(2):207-15

DIARRHEAGENIC E. COLI CHALLENGE • E. coli: most common bacterial cause of diarrhea in adults and children in developing countries • Major cause of traveller’s diarrhea • 78 different O serotypes, 34 different H serotypes • O6:H16 belongs to the most commen serotype • Most prevalent colonization factors (90%): CFA/I, CFA/II, CFA/IV Petri WA Jr et al . J Clin Invest. 2008;118(4):1277-90

STUDY DESIGN E. COLI CHALLENGE Healthy men, 18-55yr, n=30-40 per arm 28 days: two weeks run-in, one inoculation; or 49 days: two weeks run-in, two inoculations Restrictions on calcium, alcohol, medicine E. coli inoculation (1E10 CFU) at day 14 (and day 35); Stool consistency, stool frequency, quality of life, appetite, adverse events, compliance Diarrhoea, inflammation, pathogen persistence before and after challenge Inflammation & immunity before and after challenge: CRP, IP-10, serum IgA/IgG Van Hoffen et al . Manuscript submitted for publication

OUTCOMES E. COLI INFECTION INDUCES DIARRHEA • Increased total fecal output, relative fecal wet weight • Increased stool frequency, increased Bristol Stool Scale • Increased gastrointestinal complaints (Gastrointestinal Symptom Rating Scale) 100 500 7 30 % Fecal wet weight Bristol Stool Score GSRS Total fecal wet weight Percentage fecal wet weight Total fecal wet weight (g/day) 6 400 Max Bristol Stool Score GSRS total daily score 90 5 20 300 80 4 200 3 10 70 100 2 infection infection Van Hoffen et al . Manuscript 60 0 1 0 11 14 17 20 32 11 14 17 20 32 Submitted for publication Study day Study day

OUTCOMES E. COLI INFECTION INDUCES TRANSIENT INFLAMMATION • Increased fecal calprotectin • Increased serum CRP and IP-10 • Increased circulating neutrophils 8.0 10 3.0 10 4 10 3.5 Blood neutrophils (*1E9 cells/liter) Serum C-reactive protein (µg/L) Fecal calprotectin (µg/g dry) 6.0 10 3 Serum IP-10 (pg/mL) 10 2.5 10 3.0 4.0 10 2 10 2.0 10 2.5 2.0 10 1 0.0 10 1.5 10 0 10 2.0 10 15 31 1 2 3 4 13 15 21 34 10 15 17 28 10 15 17 31 Study day Study day Study day Study day

OUTCOMES E. COLI INFECTION INDUCES ANTIBODIES SPECIFIC FOR E. COLI CFA/II • Increased fecal CFA/II specific sIgA • Increased serum CFA/II specific IgG 10 4 2.0 Fecal sIgA-CFA/II (AU/mg dry) Serum IgG-CFA/II (AU/mL) 1.5 10 3 10 2 1.0 10 1 0.5 10 0 0.0 13 34 10 28 Study day Study day

E. COLI CHALLENGE MODEL: DIETARY INTERVENTIONS Intervention studies with food ingredients Milk calcium*: • % of faecal wet weight decreased on day 2 after infection Milk fat**: • Decreased symptoms on day 2 after infection: • Stool frequency • Gastro-intestinal symptoms (GSRS) Probiotics***: • Not effective in 2 studies *Bovee-Oudenhoven et al. Gastroenterology 2003;125(2):469-76. **Ten • Bruggencate et al . J Nutr. 2016;146(2):249-55. ***Ouwehand et al. B J Nutr Due to anti-inflammatory effect? 2014;111(3):465-73, Ten Bruggencate et al. Eur J Clin Nutr. 2015;69(3):385-91.

ARE CHALLENGE MODELS: FEASIBLE IN THE YOUNG AND THE OLD? Ethics Risks Biological readouts Logistics • Infants: only if no impact on health, using non-invasive procedures (unless required for other/medical reasons e.g. vaccination program) • Elderly: only if no (major) impact on health, including healthy elderly, taking vulnerability into account