FOODHYPERNES: Towards comprehensive analytical methods for partially hydrolysed gluten to assess product safety for celiac disease patients

Clinical short time challenges of intact and hydrolyzed barley gluten peptides of CeD-patients in remission Towards comprehensive analytical methods for partially hydrolyzed gluten to assess product safety for celiac disease patients Immunosafe CeD is an EU project administered by the Research Council of Norway. The project will contribute to a safer daily food life for celiac patients by highlighting key aspects of today's recommendations and increasing knowledge about a gluten-free diet. In Norway, Knut E. A. Lundin, senior physician at Oslo University Hospital and professor at the University of Oslo (UiO) and professor Ludvig M. Sollid at UiO will participate. According to European guidelines, it is assumed that an intake of 10 mg per day should be safe for a celiac patient. Research shows, however, that there are great differences in how much gluten causes symptoms and tot damage. Another important perspective is that the limit level is based on knowledge of the intake of wheat protein, and not gluten proteins from rye and barley. It is not known with certainty whether rye and barley will result in the same immunological response in this patient group. A third issue that the immunosafe-CeD project wants to shed light on is whether the analytical methods used to measure the true gluten content in "gluten-free foods" are precise enough. This applies in particular to gluten proteins such as malt extract from barley, where the protein is partially broken down into shorter chains, so-called hydrolysed proteins. Malt extract from barley is frequently used by the food industry as an additive in foods such as crackers, sauces, snacks, "ready meals" and beverages such as beer, with the intention of increasing taste and colour. The project aims to give: • Increased food safety for celiac patients • Increased knowledge about tolerance of gluten from different types of cereal • Establish new knowledge about hydrolyzed gluten proteins Increased insight into better analysis methods for the detection of gluten in food items We want to carry out a clinical study in which we test whether gluten proteins from barley, both intact proteins and malt extract, will be able to produce the same immune response in celiacs compared to consumption of wheat gluten. Testing of gluten proteins is based on the principle that consumption of these triggers an inflammatory reaction in celiacs. Activation of immune cells (CD4+ T-lymphocytes) leads to the release of a signaling substance called Interleukin-2 (IL-2). These can be measured in the blood shortly after consuming gluten, and the amount of IL-2 can be precisely quantified. Recent research has shown that IL-2 is a specific and sensitive biomarker for the intake of even low amounts of gluten and can be associated to the degree of symptoms. For the study, we need participants who: • Has been diagnosed with celiac disease (biopsy) • Has lived on a gluten-free diet for at least 24 months and is symptom-free from his celiac disease • Are sensitive to unwanted gluten intake, i.e. experience symptoms even when consuming small amounts of gluten • Are between 18-80 years old • Has a BMI of 18-30 kg/m2 The participant must otherwise be healthy and not have other known diseases (diabetes type 1, cardiovascular diseases, kidney diseases, untreated metabolic diseases, inflammatory bowel diseases, food allergies or use immunosuppressive medications. If the participant is a woman of childbearing age, she must use contraception. She must nor be pregnant or breastfeeding. The clinical study will be carried out in its entirety at the Center for Clinical Nutrition, located in the building next to the Rikshospitalet. The study will require the participant to appear physically five times. Participants must show up fasting (last 8 hours), with the exception of water that can be consumed. The first attendance (screening visit) will consist of a personal consultation with a clinical nutritionist where key aspects of the participant's health are mapped, as well as blood samples being taken so that the participant's adherence to a gluten-free diet and nutritional status are mapped. Participants will also be weighed and their height measured. At the next four visits, at 4-week intervals, varying amounts of gluten will be distributed to the participant. The study will take place blindly, so the participant does not know what he will receive from time to time. At all five attendances, blood samples will be taken and the participant will fill standardized form that maps symptoms. In terms of time, the first meeting will be approx. 1 hour duration, while the four subsequent attendances will each be of approx. 5 hours duration. Participants must give written consent to participate in the study. Participants have the right to withdraw from the study without it implying any kind of negative outcome or affecting the doctor-patient relationship.

This is a joint European project with participants from Germany, Italy and Norway that is related to celiac disease (CeD). CeD is a common food-induced inflammatory disease of the small intestine that caused by ingestion of gluten from wheat, barley and rye. The only effective treatment available is a strict lifelong gluten-free (GF) diet. GF products for CeD patients must not exceed the regulatory threshold of 20 mg/kg of gluten. Compliance of foods is typically assessed with an enzyme-linked immunosorbent assay (ELISA), but this assay performs less well in food products with fermented or partially hydrolysed gluten. The objectives of this joint European project will address this issue. In the project we will provide a comprehensive and unique toolbox of novel and validated methods to detect gluten (both intact and partially hydrolysed) in foods for CeD patients. This will be achieved by using discovery proteomics and quantitative LC-MS/MS methods, improved reference materials for partially hydrolysed gluten, CeD-patient derived monoclonal antibodies and functional gluten-specific T-cell assays. This Norwegian partner will specifically work on performing clinical challenges of CeD-patients.