Food Allergy: molecular and clinical practice

Food allergy is an adverse immunological reaction to allergens present in food. Up to 4% adults and 8% children are affected by food allergy. The increase in allergic diseases to food has led to the need for better diagnostics and more effective therapeutic approaches. This book describes the molecular biology and immunology of major food allergens, from laboratory based science to clinical immunology, encompassing novel characterisation and quantification methods, the application of recombinant food allergens in molecular diagnosis and the development of novel therapeutics. This book is the ideal reference tool for researchers, students and allergy clinicians to accurately diagnose and manage food allergies

Identification and characterisation of specific allergens of Abalone (haliotis midae; mollusca) and other seafood species

Shellfish and fish are sources of potent allergens in sensitised individuals. In view of the limited information available from published studies, especially on mollusc allergens, more detailed characterisation of the major allergens from different shellfish and fish is necessary to accurately predict cross-allergenicity. The need for a detailed characterisation of the immune response and the allergens in an indigenous mollusc species, followed a recent increase in patients presenting in the Cape Town area, with TY.pe I allergic reactions after ingestion of abalone (Haliotis midae). The first objective of this study was to determine the frequency and spectrum of reported hypersensitivity to abalone and other related seafood species in the Western Cape of South Africa. 105 volunteer subjects with suspected seafood allergy were recruited by means of a detailed seafood allergy questionnaire, advertised in the local press. The analysis of the questionnaire demonstrated clearly the importance of abalone, which was the third most frequently reported species (35%), after rock lobster and shrimp, of the 26 seafood species implicated in allergic reactions. Allergy to seafood was confirmed by the presence of specific IgE using in-house and commercial radioimmunoassays (RASTs). A novel Abalone-RAST identified specific IgE to abalone in 17/38 subjects who reported adverse reaction following the ingestion of abalone. The novel Abalone-RAST correlated positively not only with the phylogenetic closely related Snail-RAST (p<0.01) but also with squid and several indigenous crustacean species. Among subjects with multiple seafood allergies, the immune responses to molluscs species were found to be diverse. The presence of species specific allergens in the mollusc group was supported by the very low frequency of concurrent sensitivity to all mollusc species (13%) compared to concurrent sensitivity between mollusc and among the crustacea and fish group (42% and 56% respectively). The persistence of specific IgE, following a period of more than three years of seafood avoidance, was demonstrated in the mollusc species studied. The second objective was to investigate the specific immune responses to local seafood in more detail using RAST-inhibition experiments, skin prick tests (SPTs) and Western blots to demonstrate the presence of species-specific allergens. Sensitivity for the detection of abalone allergy was improved using an additional in-house SPT in six RAST negative subjects (23/38, 61 %). RAST-inhibition experiments with abalone extract demonstrated the highest degree of cross-reactivity with the Snail-RAST. In addition, a strong inhibition was achieved using commercial crustacean RASTs, confirming the presence of cross-reacting allergens in species of the same and other seafood groups. However, the low inhibitions achieved with the indigenous black mussel and squid extracts indicated that their allergen compositions differ from the species utilised in the commercial RASTs. Unique species-specific protein bands could be detected by SDS-gel electrophoresis, which clearly distinguished related mollusc species. These have not been demonstrated previously. Western blot analysis of different mollusc species identified several prominent allergens. An unexpected finding was the appearance of novel specific IgE binding reactivity after cooking the abalone. Several IgE binding proteins with similar molecular weights could be detected in immuno blots of indigenous crustacean and fish species. The next objective was to characterise the hypersensitive reactions to the local abalone species (H midae) in detail and identify the allergens found in this mollusc species. The questionnaire on abalone sensitive subjects revealed that asthma-like symptoms and the delayed onset of symptoms were frequent in sensitised individuals (42% and 34% respectively). This has previously only been reported for snail and in one case report on abalone. Surprisingly, the five abalone sensitive subjects who were studied in more detail had concurrent sensitivity to HDM and, analysed by RAST-inhibition, demonstrated two distinct types of responses. One set of subjects demonstrated a strong inhibition by HDM, indicating clearly for the first time that cross-reacting allergens or epitopes must exist between the food allergens from abalone and the air borne allergens found in HDM. SDS-gel electrophoresis demonstrated common but in addition species-specific protein bands even between very closely related abalone species from South Africa, Australia and Japan. Western blot analysis revealed two major allergens with molecular weights of about 38 and 45 kDa. Their remarkable thermal stability was demonstrated by various in-vitro and in-vivo assays. These two allergens were also present in extracts of other indigenous mollusc species, and surprisingly in some local crustacean and fish species. The individual immune responses to mollusc species were very heterogeneous for each analysed individual. The 38 kDa allergen is believed to belong to the protein family of tropomyosins, as was supported by specific IgE binding to recombinant tropomyosin of shrimp. This novel 45 kDa allergen of the South African abalone (H midae), was registered with the WHO International Union of Immunological Societies (IUIS) as Hal m 1. It is only the second allergen recognised for a mollusc species after Tod p 1 from squid. A further aim of these studies was to generate monoclonal antibodies (MoAbs) to the mollusc allergens using the hybridoma technology. The MoAbs were used, due to their consistent specific binding, to identify cross-reacting allergens among species of different seafood groups. In addition, I attempted to develop an immunologic test to distinguish between abalone species from different parts of the world. This test is of importance for the South African police to identify for forensic purposes unequivocally, fresh or processed abalone tissue exported illegally from South Africa to the Far East. Three ELISA- and Western blot assay positive clones were analysed. They demonstrated highly individual binding profiles when binding to fresh and cooked mollusc species was analysed. MoAb clones 2.11 and 2.12 (generated to the same protein of abalone) lacked complete binding to four and two of the ten abalone species respectively, allowing for very distinct species identification. Furthermore, antibody binding to cross-reacting proteins in crustacea and fish was also detected. Western blot results demonstrated clearly that these two antibodies bind to different epitopes on the same protein, making them very useful as tool for allergen and species identification. Finally, an amino acid analysis of the 3 purified antibody binding fractions of abalone was conducted. These proteins were rich in glutamine and asparagine, like tropomyosin, but differed significantly from tropomyosin with respect to serine content. The immunological findings using different patient specific sera and the monoclonal antibodies generated, provide important new information and insights into the concordant and multiple positive sensitivity to molluscs, crustacea and fish, and new information about the complexity and stability of immune responses to seafood and mollusc allergens observed in allergic subjects

Analytical methods for allergen control in food processing

Food allergy and food-related anaphylaxis have become a growing public health and food safety issue worldwide. The World Allergy Organization (WAO) estimated that 220–250 million people would live with food allergies, based on the reported prevalence of 5–8% in children and 1–2% in adults. Without a practical treatment and cure for food allergy currently available, the diligent avoidance of allergenic foods together with the prompt treatment of symptoms is the best management option available to allergic individuals. As a result, regulatory bodies in many countries mandate food allergen labelling to help allergic consumers to make informed food choices and avoid accidental exposure

Characterization of Seafood Proteins Causing Allergic Diseases

[Extract] Food allergy is increasing at a faster rate than any other allergic disorder (Gupta et al., 2007). In the last few decades, a large movement toward healthier eating makes seafood one of the major foods consumed worldwide (Wild & Lehrer, 2005). Consequently, the international trade of seafood has been growing rapidly, which reflects the popularity and frequency of consumption worldwide. The United States has become the third largest consumer of seafood in the world, with 1.86 billion kg of crustaceans in 2007 (6.04 kg/capita/year)(Food and Agriculture Organisation, 2007). Since seafood ingestion can cause severe acute hypersensitivity reactions and is recognized as one of the most common food allergies, the increased production and consumption of seafood has resulted in more frequent health problems (Lopata & Lehrer, 2009; Lopata et al., 2010). Exposure to seafood can cause a variety of health problems, including gastrointestinal disorders, urticaria, immunoglobulin E (IgE)-mediated asthma and anaphylaxis (Bang et al., 2005; Lopata & Lehrer, 2009; Malo & Cartier, 1993; Sicherer et al., 2004; Wild & Lehrer, 2005)

Relationship between serum omega-3 fatty acid and asthma endpoints

Recent studies have highlighted the potential protective role of omega-3 polyunsaturated fatty acids (n-3 PUFA) in asthma. This study aimed at determining the association between seafood intake, serum PUFA composition and clinical endpoints of asthma in adults. A cross-sectional study of 642 subjects used the European Committee Respiratory Health Survey (ECRHS) questionnaire, skin prick tests, spirometry and methacholine challenge tests following ATS guidelines. Sera was analysed for n-3 and n-6 PUFA composition. Subjects had a mean age of 34 years, were largely female (65%) and 51% were current smokers. While 99% reported fish consumption, rock lobster, mussels, squid and abalone were also consumed less frequently. The prevalence of asthma symptoms was 11%, current asthma (ECRHS definition) was 8% and non-specific bronchial hyperresponsiveness (NSBH) was much higher (26%) In adjusted models the n-3 PUFAs 20:5 (EPA) and 22:5 (DPA) were significantly associated with a decreased risk of having NSBH. Total n-3 PUFA composition was associated with decreased NSBH risk (OR = 0.92), while high n-6 PUFA composition was associated with an increased risk (OR = 1.14)

Cross-reactive epitopes and their role in food allergy

Allergenic cross-reactivity among food allergens complicates the diagnosis and management of food allergy. This can result in many patients being sensitized (having allergen-specific IgE) to foods without exhibiting clinical reactivity. Some food groups such as shellfish, fish, tree nuts, and peanuts have very high rates of cross-reactivity. In contrast, relatively low rates are noted for grains and milk, whereas many other food families have variable rates of cross-reactivity or are not well studied. Although classical cross-reactive carbohydrate determinants are clinically not relevant, α-Gal in red meat through tick bites can lead to severe reactions. Multiple sensitizations to tree nuts complicate the diagnosis and management of patients allergic to peanut and tree nut. This review discusses cross-reactive allergens and cross-reactive carbohydrate determinants in the major food groups, and where available, describes their B-cell and T-cell epitopes. The clinical relevance of these cross-reactive B-cell and T-cell epitopes is highlighted and their possible impact on allergen-specific immunotherapy for food allergy is discussed

Biocompatibility and biodegradation studies of a commercial zinc alloy for temporary mini-implant applications

In this study, the biocompatibility and in vitro degradation behaviour of a commercial zinc-based alloy (Zn-5 Al-4 Mg) were evaluated and compared with that of pure zinc for temporary orthopaedic implant applications. Biocompatibility tests were conducted using human alveolar lung epithelial cells (A549), which showed that the zinc alloy exhibits similar biocompatibility as compared to pure zinc. In vitro degradation evaluation was performed using weight loss and electrochemical methods in simulated body fluid (SBF) at 37 degrees C. Weight loss measurements revealed that the degradation of the zinc alloy was slightly lower during the initial immersion period (1-3 days), but marginally increased after 5 and 7 days immersion as compared to pure zinc. Potentiodynamic polarisation experiments showed that the zinc alloy exhibits higher degradation rate than pure zinc. However, electrochemical impedance spectroscopy analysis suggests that pure zinc is susceptible to localized degradation, whereas the zinc alloy exhibited passivation behaviour. Post-degradation analysis revealed localized degradation in both pure zinc and the zinc alloy

Conservation analysis of B-cell allergen epitopes to predict clinical cross-reactivity between shellfish and inhalant invertebrate allergens

Understanding and predicting an individual's clinical cross-reactivity to related allergens is a key to better management, treatment and progression of novel therapeutics for food allergy. In food allergy, clinical cross-reactivity is observed in patients reacting to unexpected allergen sources containing the same allergenic protein or antibody binding patches (epitopes), often resulting in severe allergic reactions. Shellfish allergy affects up to 2% of the world population and persists for life in most patients. The diagnosis of shellfish allergy is however often challenging due to reported clinical cross-reactivity to other invertebrates including mites and cockroaches. Prediction of cross-reactivity can be achieved utilizing an in-depth analysis of a few selected IgE-antibody binding epitopes. We combined available experimentally proven IgE-binding epitopes with informatics-based cross-reactivity prediction modeling to assist in the identification of clinical cross-reactive biomarkers on shellfish allergens. This knowledge can be translated into prevention and treatment of allergic diseases. To overcome the problem of predicting IgE cross-reactivity of shellfish allergens we developed an epitope conservation model using IgE binding epitopes available in the Immune Epitope Database and Analysis Resource. We applied this method to a set of four different shrimp allergens, and successfully identified several non-cross-reactive as well as cross-reactive epitopes, which have been experimentally established to cross-react. Based on these findings we suggest that this method can be used for advanced component-resolved-diagnosis to identify patients sensitized to a specific shellfish group and distinguish from patients with extensive cross-reactivity to ingested and inhaled allergens from invertebrate sources

A comprehensive review on natural bioactive compounds and probiotics as potential therapeutics in food allergy treatment

Food allergy is rising at an alarming rate and is a major public health concern. Globally, food allergy affects over 500 million people, often starting in early childhood and increasingly reported in adults. Commercially, only one approved oral immunotherapy-based treatment is currently available and other allergen-based immunotherapeutic are being investigated in clinical studies. As an alternative approach, a substantial amount of research has been conducted on natural compounds and probiotics, focusing on the immune modes of action, and therapeutic uses of such sources to tackle various immune-related diseases. Food allergy is primarily mediated by IgE antibodies and the suppression of allergic symptoms seems to be mostly modulated through a reduction of allergen-specific IgE antibodies, upregulation of blocking IgG, and downregulation of effector cell activation (e.g., mast cells) or expression of T-helper 2 (Th-2) cytokines. A wide variety of investigations conducted in small animal models or cell-based systems have reported on the efficacy of natural bioactive compounds and probiotics as potential anti-allergic therapeutics. However, very few lead compounds, unlike anti-cancer and anti-microbial applications, have been selected for clinical trials in the treatment of food allergies. Natural products or probiotic-based approaches appear to reduce the symptoms and/or target specific pathways independent of the implicated food allergen. This broad range therapeutic approach essentially provides a major advantage as several different types of food allergens can be targeted with one approach and potentially associated with a lower cost of development. This review provides a brief overview of the immune mechanisms underlying food allergy and allergen-specific immunotherapy, followed by a comprehensive collection of current studies conducted to investigate the therapeutic applications of natural compounds and probiotics, including discussions of their mode of action and immunological aspects of their disease-modifying capabilities