Crustacean allergy is a major cause of food-induced anaphylaxis. We showed previously that heating increases IgE reactivity of crustacean allergens. Here we investigate the effects of thermal ...processing of crustacean extracts on cellular immune reactivity. Raw and cooked black tiger prawn, banana prawn, mud crab and blue swimmer crab extracts were prepared and IgE reactivity assessed by ELISA. Mass spectrometry revealed a mix of several allergens in the raw mud crab extract but predominant heat-stable tropomyosin in the cooked extract. PBMC from crustacean-allergic and non-atopic control subjects were cultured with the crab and prawn extracts and proliferation of lymphocyte subsets was analysed by CFSE labelling and flow cytometry. Effector responses were assessed by intracellular IL-4 and IFN-γ, and regulatory T (CD4+CD25+CD127loFoxp3+) cell proportions in cultures were also compared by flow cytometry. For each crustacean species, the cooked extract had greater IgE reactivity than the raw (mud crab p<0.05, other species p<0.01). In contrast, there was a trend for lower PBMC proliferative responses to cooked compared with raw extracts. In crustacean-stimulated PBMC cultures, dividing CD4+ and CD56+ lymphocytes showed higher IL-4+/IFN-γ+ ratios for crustacean-allergic subjects than for non-atopics (p<0.01), but there was no significant difference between raw and cooked extracts. The percentage IL-4+ of dividing CD4+ cells correlated with total and allergen-specific IgE levels (prawns p<0.01, crabs p<0.05). Regulatory T cell proportions were lower in cultures stimulated with cooked compared with raw extracts (mud crab p<0.001, banana prawn p<0.05). In conclusion, cooking did not substantially alter overall T cell proliferative or cytokine reactivity of crustacean extracts, but decreased induction of Tregs. In contrast, IgE reactivity of cooked extracts was increased markedly. These novel findings have important implications for improved diagnostics, managing crustacean allergy and development of future therapeutics. Assessment of individual allergen T cell reactivity is required.
Allergic bronchopulmonary aspergillosis (ABPA) often presents with persistently uncontrolled asthma despite the use of corticosteroids and antifungal therapy. Omalizumab is a humanized anti-IgE ...monoclonal antibody currently used to treat severe asthma.
The aim was to assess the clinical and immunologic effects of omalizumab in ABPA in a randomized, placebo-controlled trial.
Patients with chronic ABPA were randomized to 4-month treatment with omalizumab (750 mg monthly) or placebo followed by a 3-month washout period in a cross-over design. The main endpoint was number of exacerbations. Other clinical endpoints included lung function, exhaled nitric oxide (FeNO), quality of life and symptoms. In vitro basophil activation to Aspergillus fumigatus extract and basophil FcεR1 and surface-bound IgE levels were assessed by flow cytometry.
Thirteen patients were recruited with mean total IgE 2314 ± 2125 IU/mL. Exacerbations occurred less frequently during the active treatment phase compared with the placebo period (2 vs 12 events, P = .048). Mean FeNO decreased from 30.5 to 17.1 ppb during omalizumab treatment (P = .03). Basophil sensitivity to A. fumigatus and surface-bound IgE and FcεR1 levels decreased significantly after omalizumab but not after placebo.
Omalizumab can be used safely to treat ABPA, despite high serum IgE levels. Clinical improvement was accompanied by decreased basophil reactivity to A. fumigatus and FcεR1 and surface-bound IgE levels.
Measurement of eosinophilic airway inflammation can assist in the diagnosis of allergic asthma and in the management of exacerbations, however its clinical implementation remains difficult. ...Galectin-10 has been associated with eosinophilic inflammation and has the potential to be used as a surrogate biomarker. This study aimed to assess the relationship between galectin-10 in sputum with sputum eosinophil counts, the current gold standard of eosinophil inflammation in the lung. Thirty-eight sputum samples were processed for both eosinophil counts by cytospins and semi-quantitative measurements of galectin-10 by western blots. A strong association was observed between galectin-10 levels in sputum and sputum eosinophil measurements, and they accurately determined sputum eosinophilia. The results support the potential for galectin-10 to be used as a surrogate biomarker of eosinophilic airway inflammation.
Objective: People with allergic rhinitis (AR) often self-manage in the community pharmacy setting without consulting health care professionals and trivialize their comorbidities such as asthma. A ...mobile health application (mHealth app) with a self-monitoring and medication adherence system can assist with the appropriate self-management of AR and asthma. This study aimed to identify an app effective for the self-management of AR and/or asthma.
Methods: MHealth apps retrieved from the Australian Apple App Store and Android Google Play Store were included in this study if they were developed for self-management of AR and/or asthma; in English language; free of charge for the full version; and accessible to users of the mHealth app. The mHealth app quality was evaluated on three domains using a two-stage process. In Stage 1, the apps were ranked along Domain 1 (Accessibility in both app stores). In Stage 2, the apps with Stage 1, maximum score were ranked along Domain 2 (alignment with theoretical principles of the self-management of AR and/or asthma) and Domain 3 (usability of the mHealth app using Mobile App Rating Scale instrument).
Results: Of the 418 apps retrieved, 31 were evaluated in Stage 1 and 16 in Stage 2. The MASK-air achieved the highest mean rank and covered all self-management principles except the doctor's appointment reminder and scored a total MARS mean score of 0.91/1.
Conclusions: MASK-air is ranked most highly across the assessment domains for the self-management of both AR and coexisting asthma. This mHealth app covers the majority of the self-management principles and is highly engaging.
The second data set contained 2117 allergen sequences compiled from 2 main allergen databases: the World Health Organization and International Union of Immunological Societies Allergen Nomenclature ...(http://www.allergen.org/)E2 and the Food Allergy Research and Resource Program (Version 16, http://www.allergenonline.org/).E3 Genbank accession IDs of all allergenic proteins were collected from these databases and the IDs uploaded in the Batch Entrez menu on the National Center for Biotechnology Information Web site to obtain the sequence of the protein and remove duplicate proteins. The latest distribution of protein families from the allergen data set was defined by running the hmmscan programE4 against the Pfam database (version 29.0).E5 The BLASTP program was used to align the Pacific Oyster proteins and the repertoire of known allergens using a cutoff E value of 10−7 and sequence identity of more than 50%.Gene expression analysis of potential allergens in the Pacific Oyster The expression levels of potential allergen genes were analyzed from the available RNA sequencing data from the oyster genome project.E1 The expression profiles were analyzed from 2 developmental stages (spat and juvenile) and from 10 adult organs: the adductor muscle, the digestive gland, the female gonad, the male gonad, the gill, the hemocyte, the labial palp, the outer mantle, the inner mantle, and the remaining tissue. Dried gel pieces were rehydrated with 20 ng/μL of trypsin dissolved in 40 mM ammonium bicarbonate and 10% acetonitrile for 1 hour at room temperature and subsequently incubated overnight at 37°C. The digested proteins were acidified using 0.1% formic acid and the peptides were concentrated on a SpeedVac and subjected to Liquid chromatography tandem-mass spectrometry (LC-MS/MS) analysis.Patient selection Five subjects with a convincing clinical history of allergic reactivity to shellfish and 1 nonatopic subject were recruited from the Alfred Hospital Allergy Clinic, Melbourne, Victoria, Australia. Entry Protein name Homologous allergen in the IUIS database∗ Amino acid (aa) identity (%) Overlap (aa) E value Organism Route of sensitization Source 1 B7XC66 Tropomyosin† Hel as 1 75.7 284 4.00 × 10−133 Helix aspersa (Brown garden snail) Ingestion Animal 2 K1PCV6 Triosephosphate isomerase Cra c 8 74.03 77 2.00 × 10−41 Crangon crangon (North sea shrimp) Ingestion Animal 3 K1PJ59 Triosephosphate isomerase Der f 25 73.37 169 6.00 × 10−92 Dermatophagoides farinae (HDM) Inhalation Animal 4 K1QX37 Enolase Thu a 2 72.83 357 0 Thunnus albacares (Yellowfin tuna) Ingestion Animal 5 K1Q350 Glyceraldehyde-3-phosphate dehydrogenase Tri a 34 71.21 330 7.00 × 10−173 Triticum aestivum (Wheat) Ingestion Plant 6 Q75W49 78-kDa glucose- regulated protein Cor a 10 71.04 618 0 Corylus avellana (European hazelnut) Inhalation Plant 7 K1QTC1 Paramyosin‡ — 68.59 799 0 Haliotis discus discus (Abalone) Ingestion Animal 8 K1R8R6 Fructose-bisphosphate aldolase Sal s 3 67.31 364 4.00 × 10−177 Salmo salar (Atlantic salmon) Ingestion Animal 9 K1RTQ6 Fructose-bisphosphate aldolase Sal s 3 66.67 363 1.00 × 10−178 Salmo salar (Atlantic salmon) Ingestion Animal 10 K1PNQ5 Heat shock protein HSP 90-alpha 1 Asp f 12 66.26 412 0 Aspergillus fumigatus Inhalation Fungi 11 K1QNV6 Tropomyosin† Tod p 1 65.31 271 3.00 × 10−84 Todarodes pacificus (Squid) Ingestion Animal 12 K1R266 Retinal dehydrogenase 1† Tyr p 35 61 472 1.00 × 10−170 Tyrophagus putrescentiae (Storage mite) Inhalation Animal 13 K1QNT7 Aldehyde dehydrogenase, mitochondrial Tyr p 35 60 482 1.00 × 10−180 Tyrophagus putrescentiae (Storage mite) Inhalation Animal 14 K1QVK0 Transaldolase Fus p 4 59.2 326 1.00 × 10−125 Fusarium proliferatum Inhalation Fungi 15 K1QVG5 Retinal dehydrogenase 1† Tyr p 35 59 474 1.00 × 10−169 Tyrophagus putrescentiae (Storage mite) Inhalation Fungi 16 K1PLF9 Arginine kinase Bomb m 1 59.13 345 3.00 × 10−147 Bombyx mori (Silkworm moth) Ingestion Animal 17 K1Q3F4 Inorganic pyrophosphatase Der f 32 58.24 261 1.00 × 10−113 Dermatophagoides farinae (HDM) Inhalation Animal 18 K1Q9Z4 Aldehyde dehydrogenase Tyr p 35 56 195 8.00 × 10−65 Tyrophagus putrescentiae (Storage mite) Inhalation Animal 19 K1P9D0 Stress-70 protein, mitochondrial Pen c 19 55.22 431 3.00 × 10−163 Penicillium citrinum Inhalation Fungi 20 K1QX26 Endoplasmin Asp f 12 54.04 198 9.00 × 10−61 Aspergillus fumigatus Inhalation Fungi 21 K1Q7T5 Protein disulfide-isomerase Alt a 4 52.27 44 3.00 × 10−11 Alternaria alternata Inhalation Fungi 22 K1Q5P7 Peptidyl-prolyl cis-trans isomerase Cat r 1 52.17 161 5.00 × 10−53 Catharanthus roseus (Madagascar periwinkle) Inhalation Plant 23 K1R4Z3 Malate dehydrogenase, mitochondrial Mala f 4 51.43 280 1.00 × 10−91 Malassezia furfur Inhalation Fungi 24 K1Q6X5 Protein disulfide-isomerase Alt a 4 50 68 1.00 × 10−15 Alternaria alternata Inhalation Fungi Table I Proteins identified across all 3 methods with their matched allergens source and routes of sensitization Protein Gene name Best matched allergen Amino acid identity (%) Very likely allergenic (amino acid identity ≥70%) Tropomyosin CGI_10013163 Tropomyosin (Crassostrea gigas) 92.94 Tropomyosin CGI_10013164 Tropomyosin, partial (Crassostrea virginica) 86.67 Tubulin alpha chain CGI_10002456 Der f 33 allergen (Mite) 85.71 Tubulin alpha-1C chain CGI_10002455 Der f 33 allergen (Mite) 83.2 Tubulin alpha-3 chain CGI_10018930 Der f 33 allergen (Mite) 82.43 Tubulin alpha-1C chain CGI_10024998 Der f 33 allergen (Mite) 81.8 Tubulin alpha-1C chain CGI_10007570 Der f 33 allergen (Mite) 81.53 Tubulin alpha-1C chain CGI_10024999 Der f 33 allergen (Mite) 81.35 78-kDa glucose-regulated protein CGI_10015492 Aed a 8 (Mosquito) 81 Tubulin alpha-1C chain CGI_10002454 Der f 33 allergen (Mite) 80 Tubulin alpha-1C chain CGI_10008247 Der f 33 allergen (Mite) 77.7 Tubulin alpha-1A chain CGI_10007571 Der f 33 allergen (Mite) 77.63 Fructose-bisphosphate aldolase CGI_10019801 Thu a 3 (Tuna) 74.29 Heat shock protein 70 B2 CGI_10010646 Der f 28 allergen (Mite) 74.27 Heat shock protein 70 B2 CGI_10010647 Der f 28 allergen (Mite) 74.27 Triosephosphate isomerase CGI_10003538 Triosephosphate isomerase (Shrimp) 74.03 Heat shock protein 68 CGI_10002594 Der f 28 allergen (Mite) 73.96 Triosephosphate isomerase CGI_10003539 Der f 25 allergen (Mite) 73.37 Heat shock protein 70 B2 CGI_10003417 Der f 28 allergen (Mite) 73.13 Enolase CGI_10022154 Enolase (Tunas) 72.83 Fructose-bisphosphate aldolase CGI_10025556 Thu a 3 (Tuna) 72.22 Glyceraldehyde-3-phosphate dehydrogenase CGI_10010974 Glyceraldehyde-3-phosphate dehydrogenase (Wheat) 71.21 Peptidyl-prolyl cis-trans isomerase E (PPIase E) CGI_10026365 Der f 6 (Mite) 70.19 Likely allergenic (amino acid identity ≥50%, but <70%) Ferritin CGI_10016317 Ferritin (Mite) 69.92 Paramyosin CGI_10001653 Paramyosin (Abalone) 68.59 Arginine kinase CGI_10021480 Arginine kinase (Octopus) 66.96 Arginine kinase CGI_10021483 Arginine kinase (Octopus) 66.67 Heat shock protein HSP 90-alpha 1 CGI_10017621 Asp f 12 (Fungus) 66.26 Ferritin CGI_10021660 Ferritin (Mite) 66.06 Cytochrome c CGI_10012574 Cur l 3 (Fungus) 66.02 Superoxide dismutase (Cu-Zn) CGI_10017958 Ole e 3 (Olive tree) 66 Peptidyl-prolyl cis-trans isomerase 6 CGI_10022249 Cyclophilin (Carrot) 65.96 Ferritin CGI_10027591 Ferritin (Mite) 65.58 78-kDa glucose-regulated protein CGI_10008834 Der f 28 (Mite) 65.3 78-kDa glucose-regulated protein CGI_10027395 Der f 28 (Mite) 65.25 Arginine kinase CGI_10024056 Arginine kinase (Octopus) 64.65 Inorganic pyrophosphatase CGI_10027722 Der f 32 (Mite) 64.24 Fructose-bisphosphate aldolase CGI_10000078 Sal s 3 (Salmon) 64.12 60S ribosomal protein L3 CGI_10010529 Asp f 23 (Fungus) 63.96 Arginine kinase CGI_10021482 Arginine kinase (Octopus) 63.83 Heat shock protein 68 CGI_10002823 Der f 28 (Mite) 63.34 40-kDa peptidyl-prolyl cis-trans isomerase CGI_10015504 Cyclophilin (Carrot) 62.72 Peptidyl-prolyl cis-trans isomerase CGI_10013880 Cyclophilin (Carrot) 61.76 Plasma kallikrein CGI_10016607 Der f 3 (Mite) 61.7 60S ribosomal protein L3 CGI_10012282 Asp f 23 (Fungus) 60.76 Arginine kinase CGI_10021481 Arginine kinase (Octopus) 60.6 Tubulin alpha chain CGI_10018903 Der f 33 (Mite) 60.49 78-kDa glucose-regulated protein CGI_10018425 Der f 28 (Mite) 59.9 Eukaryotic translation initiation factor 3 subunit I (fragment) CGI_10025943 For t 1 (Midges) 59.88 Calcium-binding atopy-related autoantigen 1 CGI_10026057 Hom s 4 (Human) 59.3 Transaldolase CGI_10002311 Fus p 4 (Fungus) 59.2 Peptidyl-prolyl cis-trans isomerase B CGI_10023851 Cyclophilin (Carrot) 58.7 Peptidyl-prolyl cis-trans isomerase CGI_10023850 Asp f 27 (Fungus) 58.33 Aldehyde dehydrogenase, mitochondrial CGI_10012671 Cla h 10 (Fungus) 58.02 Thioredoxin CGI_10021611 Mala s 13 (Yeast) 58 Peptidyl-prolyl cis-trans isomerase CGI_10024975 Cyclophilin (Carrot) 57.63 Thioredoxin domain-containing protein 5 CGI_10009327 Alt a 4 (Fungus) 57.45 60S acidic ribosomal protein P1 CGI_10009326 Alt a 12 (Fungus) 57.14 NK-tumor recognition protein CGI_10007438 Asp f 27 (Fungus) 56.8 Thaumatin-like protein 1a CGI_10012508 Pathogenesis related protein 5 (Apple) 55.62 Superoxide dismutase CGI_10017307 Pis v 4 (Pistachio) 55.61 Peptidyl-prolyl cis-trans isomerase CGI_10006179 Asp f 11 (Fungus) 55.56 Peptidylprolyl isomerase domain and WD repeat-containing protein 1 CGI_10011521 Mala s 6 (Yeast) 55.47 Stress-70 protein, mitochondrial CGI_10016162 Pen c 19 (Fungus) 55.22 Peptidyl-prolyl cis-trans isomerase-like 6 CGI_10024382 Cat r 1 (Periwinkle) 55.22 U4/U6.U5 tri-snRNP-associated protein 1 CGI_10021218 Hom s 1 (Human) 54.96 Alpha-amylase CGI_10022190 Bla g 11 (Cockroach) 54.37 Alpha-amylase CGI_10022189 Bla g 11 (Cockroach) 54.18 Endoplasmin CGI_10025730 Asp f 12 (Fungus) 54.04 Calmodulin CGI_10006247 Amb a 9 (Ragweed) 53.7 Calmodulin CGI_10014525 B1 protein allergen (Bermuda grass) 53.57 Aldehyde dehydrogenase CGI_10021688 Cla h 10 (Fungus) 52.82 Retinal dehydrogenase 1 CGI_10026868 Cla h 10 (Fungus) 52.33 Protein disulfide-isomeras
Allergic diseases are the most common chronic immune‐mediated disorders and can manifest with an enormous diversity in clinical severity and symptoms. Underlying mechanisms for the adverse immune ...response to allergens and its downregulation by treatment are still being revealed. As a result, there have been, and still are, major challenges in diagnosis, prediction of disease progression/evolution and treatment. Currently, the only corrective treatment available is allergen immunotherapy (AIT). AIT modifies the immune response through long‐term repeated exposure to defined doses of allergen. However, as the treatment usually needs to be continued for several years to be effective, and can be accompanied by adverse reactions, many patients face difficulties completing their schedule. Long‐term therapy also potentially incurs high costs. Therefore, there is a great need for objective markers to predict or to monitor individual patient's beneficial changes in immune response during therapy so that efficacy can be identified as early as possible. In this review, we specifically address recent technical developments that have generated new insights into allergic disease pathogenesis, and how these could potentially be translated into routine laboratory assays for disease monitoring during AIT that are relatively inexpensive, robust and scalable.
The potential of precision medicine in allergy and asthma has only started to be explored. A significant clarification in the pathophysiology of rhinitis, chronic rhinosinusitis, asthma, food allergy ...and drug hypersensitivity was made in the last decade. This improved understanding led to a better classification of the distinct phenotypes and to the discovery of new drugs such as biologicals, targeting phenotype‐specific mechanisms. Nevertheless, many conditions remain poorly understood such as non‐eosinophilic airway diseases or non‐IgE–mediated food allergy. Moreover, there is a need to predict the response to specific therapies and the outcome of drug and food provocations. The identification of patients at risk of progression towards severity is also an unmet need in order to establish adequate preventive or therapeutic measures. The implementation of precision medicine in the clinical practice requires the identification of phenotype‐specific markers measurable in biological matrices. To become useful, these biomarkers need to be quantifiable by reliable systems, and in samples obtained in an easy, rapid and cost‐efficient way. In the last years, significant research resources have been put in the identification of valid biomarkers for asthma and allergic diseases. This review summarizes these recent advances with focus on the biomarkers with higher clinical applicability.
Shellfish allergy is a major cause of food-induced anaphylaxis, but the allergens are not well characterized. This study examined the effects of heating on blue swimmer crab (Portunus pelagicus) ...allergens in comparison with those of black tiger prawn (Penaeus monodon) by testing reactivity with shellfish-allergic subjects' serum IgE. Cooked extracts of both species showed markedly increased IgE reactivity by ELISA and immunoblotting, and clinical relevance of IgE reactivity was confirmed by basophil activation tests. Inhibition IgE ELISA and immunoblotting demonstrated cross-reactivity between the crab and prawn extracts, predominantly due to tropomyosin, but crab-specific IgE-reactivity was also observed. The major blue swimmer crab allergen tropomyosin, Por p 1, was cloned and sequenced, showing strong homology with tropomyosin of other crustacean species but also sequence variation within known and predicted linear IgE epitopes. These findings will advance more reliable diagnosis and management of potentially severe food allergy due to crustaceans.
Background Synthetic peptide immunoregulatory epitopes are a new class of immunotherapy to treat allergic rhinoconjunctivitis (ARC). Grass allergen peptides, comprising 7 synthetic T-cell epitopes ...derived from Cyn d 1, Lol p 5, Dac g 5, Hol l 5, and Phl p 5, is investigated for treatment of grass pollen–induced ARC. Objective We sought to evaluate the efficacy, safety, and tolerability of intradermally administered grass allergen peptides. Methods A multicenter, randomized, double-blind, placebo-controlled study evaluated 3 regimens of grass allergen peptides versus placebo in patients with grass pollen–induced allergy (18-65 years). After a 4-day baseline challenge to rye grass in the environmental exposure unit (EEU), subjects were randomized to receive grass allergen peptides at 6 nmol at 2-week intervals for a total of 8 doses (8x6Q2W), grass allergen peptides at 12 nmol at 4-week intervals for a total of 4 doses (4x12Q4W), or grass allergen peptides at 12 nmol at 2-week intervals for a total of 8 doses (8x12Q2W) or placebo and treated before the grass pollen season. The primary efficacy end point was change from baseline in total rhinoconjunctivitis symptom score across days 2 to 4 of a 4-day posttreatment challenge (PTC) in the EEU after the grass pollen season. Secondary efficacy end points and safety were also assessed. Results Two hundred eighty-two subjects were randomized. Significantly greater improvement (reduction of total rhinoconjunctivitis symptom score from baseline to PTC) occurred across days 2 to 4 with grass allergen peptide 8x6Q2W versus placebo (−5.4 vs −3.8, respectively; P = .0346). Greater improvement at PTC also occurred for grass allergen peptide 8x6Q2W versus placebo ( P = .0403) in patients with more symptomatic ARC. No safety signals were detected. Conclusion Grass allergen peptide 8x6Q2W significantly improved ARC symptoms after rye grass allergen challenge in an EEU with an acceptable safety profile.
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