In-depth characterization of the serum antibody epitope repertoire in inflammatory bowel disease using phage-displayed immunoprecipitation sequencing

Inflammatory bowel diseases (IBD), e.g. Crohn's disease (CD) and ulcerative colitis (UC), are chronic immune-mediated inflammatory diseases. A comprehensive overview of an IBD-specific antibody epitope repertoire is, however, lacking. We leveraged a high-throughput phage-displayed immunoprecipitation sequencing (PhIP-seq) workflow to identify antibodies against 344,000 antimicrobial, immune and food antigens in 497 IBD patients as compared to 1,326 controls. IBD was characterized by 373 differentially abundant antibodies (202 overrepresented and 171 underrepresented), with 17% shared by both IBDs, 55% unique to CD and 28% unique to UC. Antibodies against bacterial flagellins dominated in CD and were associated with ileal involvement, fibrostenotic disease and anti-Saccharomyces cerevisiae antibody positivity, but not with fecal microbiome composition. Antibody epitope repertoires accurately discriminated CD from controls (AUC=0.89), and similar discrimination was achieved when using only ten antibodies (AUC=0.87). IBD patients thus show a distinct antibody repertoire against selected peptides, allowing patient stratification and discovery of immunological targets.

Increased Frequency of CTLA-4 and PD-1 Expressing Regulatory T Cells and Basophils With an Activating Profile in Infants With Moderate-to-Severe Atopic Dermatitis Hypersensitized to Food Allergens

Background: Infants with severe atopic dermatitis (AD) may be sensitized to foods that have not been introduced into their diet, posing a risk for developing an immediate hypersensitivity reaction on the first exposure to the food to which they are sensitized. The aim of this work was to perform an analysis of the sensitization profile in infants with moderate-to-severe AD and to identify cellular and molecular markers for food allergy (FA).Methods: Blood samples from healthy donors and children with moderate-to-severe AD were studied. Specific IgE to several allergens were determined using ImmunoCAP FEIA system and ISAC technology. Furthermore, using flow cytometry-based studies, basophils and regulatory T (Treg) cells were phenotypically characterized.Results: 90% of children with AD were sensitized to food antigens before introducing them into the diet, and 100% developed FA. Phenotypic analysis showed a significantly higher percentage of CTLA-4 and PD-1 expressing Treg cells in AD patients than in healthy controls. Basophils from patients exhibited a marked reduction in the expression of CD300a, higher expression of FcεRI and CXCR4, and to some extent higher expression of CD63 and CD300c.Conclusions: Infants with moderate-to-severe AD are at high risk of being sensitized to food allergens. Therefore, to avoid allergic reactions, broad-spectrum sensitization studies are necessary before introducing complementary diet. Increased expression of CTLA-4 and PD-1 suggests greater suppressive potential of Treg cells in infants with AD than healthy controls. Furthermore, our results suggest a role for CD300 molecules on circulating basophils as possible biomarkers for FA susceptibility.

Determination of Specific IgG to Identify Possible Food Intolerance in Athletes Using ELISA

Nutrition is considered one of the foundations of athletic performance, and post-workout nutritional recommendations are fundamental to the effectiveness of the recovery and adaptive processes. Therefore, at present, new directions in dietetics are being formed, focused on the creation of personalized diets. To identify the probable risk of somatic and allergic reactions upon contact with food antigens, we used the method of enzyme-linked immunosorbent assay (ELISA) for the quantitative determination of IgG antibodies in the blood plasma of athletes against protein–peptide antigens accommodated in food. The study enrolled 40 athletes of boating and fighting sport disciplines. We found that the majority of the studied participants were characterized by an elevated IgG level against one or two food allergens (barley, almond, strawberry, etc.). Comparative analysis of the semiquantitative levels of IgG antibodies in athletes engaged in boating and fighting did not reveal significant differences between these groups. As a result, foods that are likely to cause the most pronounced immune response amongst the studied participants can be identified, which may indicate the presence of food intolerances. An athlete’s diet is influenced by both external and internal factors that can reduce or worsen the symptoms of a food intolerance/allergy associated with exercise. The range of foods is wide, and the effectiveness of a diet depends on the time, the place, and environmental factors. Therefore, during the recovery period (the post-competition period), athletes are advised to follow the instructions of doctors and nutritionists. An effective, comprehensive recovery strategy during the recovery period may enhance the adaptive response to fatigue, improving muscle function and increasing exercise tolerance. The data obtained may be useful for guiding the development of a new personalized approach and dietary recommendations covering the composition of athletes’ diet and the prevalence of food intolerance.

Genetic regulation of serum IgA levels and susceptibility to common immune, infectious, kidney, and cardio-metabolic traits

Immunoglobulin A (IgA) mediates mucosal responses to food antigens and the intestinal microbiome and has a known role in susceptibility to mucosal pathogens, celiac disease, inflammatory bowel disease, and IgA nephropathy. We performed genetic analyses of serum IgA levels in 41,263 individuals of diverse ancestries. We observed unexpected variability in IgA levels across major ancestral populations, with African ancestry being reproducibly associated with higher serum IgA levels compared to other ancestries. The trans-ethnic GWAS analysis identified 20 genome-wide significant loci associated with serum IgA levels, including nine known and 11 novel loci. Systematic co-localization analysis with blood and primary immune cell expression QTLs prioritized candidate genes for 14 of 20 loci. Most GWAS loci encoded genes that produce immune defects and IgA abnormalities when genetically manipulated in mice. We uncovered positive genetic correlations of serum IgA levels with IgA nephropathy, type 2 diabetes and body mass index, as well as negative genetic correlations with celiac disease, inflammatory bowel disease, several infections, and intestinal microbiome diversity. Our findings provide novel insights into the genetic regulation of IgA production and its potential role in human disease.

Eosinophilic Esophagitis: Atopy Conditions of the Esophagus

Eosinophilic esophagitis is an immune-allergic pathology of multifactorial etiology(genetic and environmental) characterized by major symptoms of esophagealdysphagia and eosinophil-predominant inflammation of the esophageal mucosathat affects both pediatric and adult patients. EoE is an immune-mediated diseaseby which environmental and food antigens stimulate the Th2 inflammatorycascade. It is correlated with food allergy and atopy condition such as asthma, atopydermatitis, rhinitis allergic and often in conjunction with Gastroesophageal RefluxDisease (GERD). Eosinophilic esophagitis (EoE) was first described in the 1990s,showing an increasing incidence and prevalence since then, in the United States isestimated to be approximately 57 per 100,000 persons being the leading cause offood impaction and the major cause of dysphagia. Its symptoms, which includeheartburn, regurgitation, and esophageal stenosis. This symptomps similar to thoseof gastroesophageal reflux disease, causing delays in diagnosis and treatment. Theendoscopic findings such as furrows, esophageal mucosa trachealization, andwhitish exudates, this diagnosis should be confirmed histologically confirmed bybiopsy on the presence of more than 15 eosinophils per high-power field and theexclusion of other causes of eosinophilia. Management includes medications, diet,and surgical dilatation.

Diet Diversity Through the Life-Course as an Opportunity Toward Food Allergy Prevention

The prevalence of food allergies (FA) is increasing worldwide. Generally, the onset of allergies, including FA, begins in early childhood and may persist and/or develop through the life-course. Even though epidemiological studies have focused mainly on children, allergies can also occur for the first-time during adulthood. Within the prolongation of life, it is expected that allergies will be encountered more often even in older people. Recent findings suggest that an early exposure to diverse food antigens may promote the development of immune tolerance. Accordingly, diet diversity during the first year of life or even earlier may have a positive impact on the prevention of allergies. The anti-inflammatory properties of some dietary nutrients may positively contribute to a tolerogenic immune environment too. Diet diversity is associated with a more favorable microbiome, and increasing evidence suggests a promising role of gut microbiota manipulation in inducing immune tolerance. Unjustified avoidance of allergenic foods may expose to intakes of some nutrients below recommended levels through the life-course, even more in cases of self-diagnosis and treatment of presumed forms of food intolerance. Nutritional strategies including the early exposure to a variety of food antigens are a promising area of research for preventive purposes through the life-course possibly extending positive outcomes to older stages. The aim of this paper is to highlight the role of diet diversity in preventing the development of FA starting in early life, as well as to provide an overview of the main strategies to prevent related nutritional issues throughout the life-course.

Retinoic Acid Induces Functionally Suppressive Foxp3+RORγt+ T Cells In Vitro

IntroductionCD4+ T cells with regulatory function co-expressing Foxp3 and RORγt are linked to the development of oral tolerance towards innocuous food antigens in mice. This study aimed to discern the role played by IL-6 and retinoic acid (RA) in the in vitro generation of Foxp3+RORγt+ T cells and to investigate whether such cells have suppressive properties.MethodsCD4+CD25- T cells isolated from the spleen of BALB/c mice, were stimulated in the presence of IL-2 alone or together with TFG-β and different concentrations of IL-6 and/or RA. Percentage of Foxp3+, RORγt+, IL-17+, Foxp3+RORγt-, Foxp3+RORγt+, and Foxp3-RORγt+ T cells within the total CD4+ T cell population, production of cytokines (IL-10 and IL-17A) and gene expression (Foxp3, Rorc, Tgfb1, Il6, Il10, and Il17) were assessed at different time points. The phenotype and ability of cells generated from CD4+CD44-CD62L+ cells in the presence of RA to suppress effector T cell proliferation was assessed.ResultsTGF-β plus IL-6 induced the generation of Foxp3+ and double positive Foxp3+RORγt+ T cells to a higher extent than TGF-β alone at the beginning of the incubation period, although expression of Foxp3 subsequently declined. RA, added to TGF-β, increased Foxp3 and Rorc expression and Foxp3 and RORγt transcription and promoted the differentiation of Foxp3+RORγt- and Foxp3+RORγt+ cells that expressed and secreted IL-17. Foxp3+ T cells generated in vitro in presence of RA were functionally suppressive.ConclusionsUnder the influence of IL-2 and TGF-β, suppressive Foxp3+RORγt+ T cells that express and secrete IL-17 can be produced in vitro and RA further contributes to stabilize this phenotype.

Intestinal Regulatory T Cells as Specialized Tissue-Restricted Immune Cells in Intestinal Immune Homeostasis and Disease

FOXP3+ regulatory T cells (Treg cells) are a specialized population of CD4+ T cells that restrict immune activation and are essential to prevent systemic autoimmunity. In the intestine, the major function of Treg cells is to regulate inflammation as shown by a wide array of mechanistic studies in mice. While Treg cells originating from the thymus can home to the intestine, the majority of Treg cells residing in the intestine are induced from FOXP3neg conventional CD4+ T cells to elicit tolerogenic responses to microbiota and food antigens. This process largely takes place in the gut draining lymph nodes via interaction with antigen-presenting cells that convert circulating naïve T cells into Treg cells. Notably, dysregulation of Treg cells leads to a number of chronic inflammatory disorders, including inflammatory bowel disease. Thus, understanding intestinal Treg cell biology in settings of inflammation and homeostasis has the potential to improve therapeutic options for patients with inflammatory bowel disease. Here, the induction, maintenance, trafficking, and function of intestinal Treg cells is reviewed in the context of intestinal inflammation and inflammatory bowel disease. In this review we propose intestinal Treg cells do not compose fixed Treg cell subsets, but rather (like T helper cells), are plastic and can adopt different programs depending on microenvironmental cues.

Update on the Role of Allergy in Pediatric Functional Abdominal Pain Disorders: A Clinical Perspective

Both functional abdominal pain disorders (FAPDs) and food allergies are relatively common in children and adolescents, and most studies report an association between FAPDs and allergic conditions. FAPDs share pathophysiologic processes with allergies, including both immune and psychological processes interacting with the microbiome. No conclusive data are implicating IgE-mediated reactions to foods in FAPDs; however, there may be patients who have IgE reactions localized to the gastrointestinal mucosa without systemic symptoms that are not identified by common tests. In FAPDs, the data appears stronger for aeroallergens than for foods. It also remains possible that food antigens initiate an IgG reaction that promotes mast cell activation. If a food allergen is identified, the management involves eliminating the specific food from the diet. In the absence of systemic allergic symptoms or oral allergy syndrome, it appears unlikely that allergic triggers for FAPDs can be reliably identified by standard testing. Medications used to blunt allergic reactions or symptomatically treat allergic reactions may be useful in FAPDs. The purpose of the current manuscript is to review the current literature regarding the role of allergy in FAPDs from a clinical perspective, including how allergy may fit in the current model of FAPDs.