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Автор: Grafiati

Опубліковано: 21 вересня 2024

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1

Sokolov, Pavel, Irina Evsegneeva, Alexander Karaulov, Alyona Sukhanova, and Igor Nabiev. "Allergen Microarrays and New Physical Approaches to More Sensitive and Specific Detection of Allergen-Specific Antibodies." Biosensors 14, no. 7 (July 20, 2024): 353. http://dx.doi.org/10.3390/bios14070353.

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The prevalence of allergic diseases has increased tremendously in recent decades, which can be attributed to growing exposure to environmental triggers, changes in dietary habits, comorbidity, and the increased use of medications. In this context, the multiplexed diagnosis of sensitization to various allergens and the monitoring of the effectiveness of treatments for allergic diseases become particularly urgent issues. The detection of allergen-specific antibodies, in particular, sIgE and sIgG, is a modern alternative to skin tests due to the safety and efficiency of this method. The use of allergen microarrays to detect tens to hundreds of allergen-specific antibodies in less than 0.1 mL of blood serum enables the transition to a deeply personalized approach in the diagnosis of these diseases while reducing the invasiveness and increasing the informativeness of analysis. This review discusses the technological approaches underlying the development of allergen microarrays and other protein microarrays, including the methods of selection of the microarray substrates and matrices for protein molecule immobilization, the obtainment of allergens, and the use of different types of optical labels for increasing the sensitivity and specificity of the detection of allergen-specific antibodies.

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2

Klimek, L., D. Vetter, L. von Bernus, and C. Thorn. "Allergen-Microarrays für die molekulare Komponentendiagnostik von Typ-I-Allergien." HNO 59, no. 10 (December 22, 2010): 988–93. http://dx.doi.org/10.1007/s00106-010-2224-5.

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3

Romantowski, Jan, Aleksandra Górska, Grażyna Moszkowska, Julia Kulczycka, Karolina Minkowska, Agata Rolewicz, Marita Nittner-Marszalska, and Marek Niedoszytko. "Atopy and Multisensitizations in Specific IgE Microarrays and Their Impact on Severe Asthma." Life 12, no. 10 (September 29, 2022): 1520. http://dx.doi.org/10.3390/life12101520.

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(1) Asthma is a chronic inflammatory airway disease. Around 3–10% of patients experience severe refractory asthma. These patients with high symptom intensity and frequent exacerbations present a challenge for allergologists. Their allergic vs. non-allergic profile might be different from the standard asthmatic group and this difference is vital in qualifying for anti-IgE biologicals. The aim of the study was to analyze multiple sensitizations in patients with severe asthma and assess their impact on the course of the disease. (2) Forty-two patients with severe asthma according to GINA were enrolled. They experienced at least two exacerbations during the past year and had uncontrolled asthma despite high inhaled steroid use. A microarray serum Alex test (allergen-specific IgE to 295 extracts and components) was performed together with Complete Blood Count tests, the Asthma Control Questionnaire (ACQ), the Mini Asthma Quality of Life Questionnaire (MiniAQLQ), and spirometry. (3) There were 29 female and 13 male patients. The patient mean age was 50.4 (22–70). In 25 (60%) patients, inhalant sensitizations were detected. In 9 (21%) cases, a new perennial allergen was discovered that might enable anti-IgE treatment in the future. In the entire studied group, 8 patients (19%) would still not qualify for anti-IgE, anti-IL4, or anti-IL5 treatment. A linear regression analysis revealed that a Canis familiaris allergen (Can f 1) correlated with worse asthma control in ACQ. An Aspergillus allergen (Asp f 6) correlated negatively with Forced Expiratory Volume in one second (FEV1). (4) The study presents the usefulness of the ALEX test in 21% of patients with severe asthma in qualification for anti-IgE treatment. It highlights the impact of canine and Aspergillus sensitizations on worse control in patients with severe asthma.

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4

Cretich, Marina, Daniela Breda, Francesco Damin, Marta Borghi, Laura Sola, Selim M. Unlu, Samuele E. Burastero, and Marcella Chiari. "Allergen microarrays on high-sensitivity silicon slides." Analytical and Bioanalytical Chemistry 398, no. 4 (August 22, 2010): 1723–33. http://dx.doi.org/10.1007/s00216-010-4077-x.

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5

Jeon, Hyunjin, Joo Hyun Jung, Yoonji Kim, B.S., Youngeun Kwon, and Seon Tae Kim. "Allergen Microarrays forIn VitroDiagnostics of Allergies: Comparison with ImmunoCAP and AdvanSure." Annals of Laboratory Medicine 38, no. 4 (July 28, 2018): 338–47. http://dx.doi.org/10.3343/alm.2018.38.4.338.

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6

Szameit, Sandra, Elisabeth Weber, and Christa Noehammer. "DNA microarrays provide new options for allergen testing." Expert Review of Molecular Diagnostics 9, no. 8 (November 2009): 843–50. http://dx.doi.org/10.1586/erm.09.63.

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7

Baumgart, Karl. "The whether, whither or wither of allergen microarrays." Pathology 56 (February 2024): S31. http://dx.doi.org/10.1016/j.pathol.2023.12.114.

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8

Romantowski, Jan, Aleksandra Górska, Marek Niedoszytko, Theo Gulen, Marta Gruchała-Niedoszytko, Bogusław Nedoszytko, Magdalena Lange, et al. "A Challenge for Allergologist: Application of Allergy Diagnostic Methods in Mast Cell Disorders." International Journal of Molecular Sciences 22, no. 3 (February 1, 2021): 1454. http://dx.doi.org/10.3390/ijms22031454.

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Primary and secondary mast cell activation syndromes (MCAS) can occur in patients with mastocytosis. During the past few years our knowledge about the pathogenesis and disease-triggering mechanisms in MCAS and mastocytosis have increased substantially. Whereas mastocytosis is characterized by an accumulation of neoplastic (clonal) mast cells (MC) in various organ systems, MCAS is defined by a massive and systemic activation of these cells. Mast cells are crucial effector cells in allergic diseases, thus their elevated number and activation can cause severe anaphylactic reactions and MCAS in patients with mastocytosis. However, these cells may also degranulate spontaneously or degranulate in response to non-allergic triggers leading to clinical symptoms. In mastocytosis patients, such symptoms may lead to the diagnosis of a primary MCAS. The diagnosis of a concomitant allergy in mastocytosis patients is challenging. In these patients, a mixed form (primary and secondary) of MCAS may be diagnosed. These patients may also suffer from life-threatening anaphylactic reactions when exposed to allergens. In these cases, the possibility of severe side effects of in vivo provocations can sometimes also limit diagnostic evaluations. In the current article, we discuss the diagnosis and management of patients suffering from mastocytosis and concomitant MCAS, with special emphasis on novel diagnostic tests and management, including allergen microarrays, recombinant allergen analysis, basophil activation tests, optimal prophylaxis, and specific therapies.

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9

Chinnasamy, Thiruppathiraja, Loes I. Segerink, Mats Nystrand, Jesper Gantelius, and Helene Andersson Svahn. "Point-of-Care Vertical Flow Allergen Microarray Assay: Proof of Concept." Clinical Chemistry 60, no. 9 (September 1, 2014): 1209–16. http://dx.doi.org/10.1373/clinchem.2014.223230.

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Abstract BACKGROUND Sophisticated equipment, lengthy protocols, and skilled operators are required to perform protein microarray-based affinity assays. Consequently, novel tools are needed to bring biomarkers and biomarker panels into clinical use in different settings. Here, we describe a novel paper-based vertical flow microarray (VFM) system with a multiplexing capacity of at least 1480 microspot binding sites, colorimetric readout, high sensitivity, and assay time of <10 min before imaging and data analysis. METHOD Affinity binders were deposited on nitrocellulose membranes by conventional microarray printing. Buffers and reagents were applied vertically by use of a flow controlled syringe pump. As a clinical model system, we analyzed 31 precharacterized human serum samples using the array system with 10 allergen components to detect specific IgE reactivities. We detected bound analytes using gold nanoparticle conjugates with assay time of ≤10 min. Microarray images were captured by a consumer-grade flatbed scanner. RESULTS A sensitivity of 1 ng/mL was demonstrated with the VFM assay with colorimetric readout. The reproducibility (CV) of the system was <14%. The observed concordance with a clinical assay, ImmunoCAP, was R2 = 0.89 (n = 31). CONCLUSIONS In this proof-of-concept study, we demonstrated that the VFM assay, which combines features from protein microarrays and paper-based colorimetric systems, could offer an interesting alternative for future highly multiplexed affinity point-of-care testing.

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10

Cretich, Marina, Gabriele Di Carlo, Cinzia Giudici, Sven Pokoj, Iris Lauer, Stephan Scheurer, and Marcella Chiari. "Detection of allergen specific immunoglobulins by microarrays coupled to microfluidics." PROTEOMICS 9, no. 8 (April 2009): 2098–107. http://dx.doi.org/10.1002/pmic.200800651.

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11

Fall, Barbara I., Bernadette Eberlein-König, Heidrun Behrendt, Reinhard Niessner, Johannes Ring, and Michael G. Weller. "Microarrays for the Screening of Allergen-Specific IgE in Human Serum." Analytical Chemistry 75, no. 3 (February 2003): 556–62. http://dx.doi.org/10.1021/ac026016k.

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12

Foncy, Julie, Erwan Crestel, Jean-Philippe Borges, Aurore Estève, Jean Christophe Cau, Christophe Vieu, Laurent Malaquin, and Emmanuelle Trévisiol. "Reversible magnetic clamp of a microfluidic interface for the seric detection of food allergies on allergen microarrays." Microelectronic Engineering 158 (June 2016): 16–21. http://dx.doi.org/10.1016/j.mee.2016.03.005.

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13

Tuppo, Lisa, Ivana Giangrieco, Maurizio Tamburrini, Claudia Alessandri, Adriano Mari, and Maria Antonietta Ciardiello. "Detection of Allergenic Proteins in Foodstuffs: Advantages of the Innovative Multiplex Allergen Microarray-Based Immunoassay Compared to Conventional Methods." Foods 11, no. 6 (March 19, 2022): 878. http://dx.doi.org/10.3390/foods11060878.

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Several factors can affect the allergen content and profile of a specific food, including processing procedures often leading to a decrease in allergenicity, although no change, or even an increase, have also been reported. Evaluation of the effectiveness of a processing procedure requires the availability of reliable methodologies to assess the variation in molecules able to induce allergic reactions in the analyzed food. Conventional and innovative strategies and methodologies can be exploited to identify allergenic proteins in foodstuffs. However, depending on the specific purposes, different methods can be used. In this review, we have critically reviewed the advantages of an innovative method, the multiplex allergen microarray-based immunoassay, in the detection of allergens in foodstuffs. In particular, we have analyzed some studies reporting the exploitation of an IgE-binding inhibition assay on multiplex allergen biochips, which has not yet been reviewed in the available literature. Unlike the others, this methodology enables the identification of many allergenic proteins, some of which are still unknown, which are recognized by IgE from allergic patients, with a single test. The examined literature suggests that the inhibition test associated with the multiplex allergen immunoassay is a promising methodology exploitable for the detection of IgE-binding proteins in food samples.

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14

Bacarese-Hamilton, Tito, Letizia Mezzasoma, Colin Ingham, Andrea Ardizzoni, Ruggero Rossi, Francesco Bistoni, and Andrea Crisanti. "Detection of Allergen-specific IgE on Microarrays by Use of Signal Amplification Techniques." Clinical Chemistry 48, no. 8 (August 1, 2002): 1367–70. http://dx.doi.org/10.1093/clinchem/48.8.1367.

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15

Canonica, G. Walter, Giovanni Passalacqua, and Gianni Melioli. "Poster 1001: ALLERGENIUS®: an expert system for the interpretation of allergen microarrays." World Allergy Organization Journal 7 (2014): P2. http://dx.doi.org/10.1186/1939-4551-7-s1-p2.

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16

Prosperi, Mattia C. F., Danielle Belgrave, Iain Buchan, Angela Simpson, and Adnan Custovic. "Challenges in interpreting allergen microarrays in relation to clinical symptoms: A machine learning approach." Pediatric Allergy and Immunology 25, no. 1 (October 16, 2013): 71–79. http://dx.doi.org/10.1111/pai.12139.

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17

Wiltshire, Steve, Shawn O’Malley, Jeremy Lambert, Kari Kukanskis, David Edgar, Stephen F. Kingsmore, and Barry Schweitzer. "Detection of Multiple Allergen-specific IgEs on Microarrays by Immunoassay with Rolling Circle Amplification." Clinical Chemistry 46, no. 12 (December 1, 2000): 1990–93. http://dx.doi.org/10.1093/clinchem/46.12.1990.

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18

Harris, Jennifer, Daniel E. Mason, Jun Li, Keith W. Burdick, Bradley J. Backes, Teresa Chen, Aaron Shipway, et al. "Activity Profile of Dust Mite Allergen Extract Using Substrate Libraries and Functional Proteomic Microarrays." Chemistry & Biology 11, no. 10 (October 2004): 1361–72. http://dx.doi.org/10.1016/j.chembiol.2004.08.008.

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19

Ott, Hagen, Regina Fölster-Holst, Hans F. Merk, and Jens Malte Baron. "Allergen microarrays: a novel tool for high-resolution IgE profiling in adults with atopic dermatitis." European Journal of Dermatology 20, no. 1 (January 2010): 054–61. http://dx.doi.org/10.1684/ejd.2010.0810.

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20

Garriga-Baraut, Teresa, Moises Labrador-Horrillo, Mercé Tena, Concepción De Linares, Olga Esteso-Hontoria, Carlos Pedemonte, Maria Basagaña-Torrento, et al. "A real-life ImmunoCAT study: impact of molecular diagnosis through ImmunoCAPTM ISAC 112 on immunotherapy prescription in pollen-polysensitized patients in Catalonia, Spain." Allergologia et Immunopathologia 52, no. 4 (July 1, 2024): 21–29. http://dx.doi.org/10.15586/aei.v52i4.1077.

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Background: Molecular diagnosis in allergology helps to identify multiple allergenic molecules simultaneously. The use of purified and/or recombinant allergens increases the accuracy of individual sensitization profiles in allergic patients. Objective: To assess the impact of molecular diagnosis through the ImmunoCAPTM ISAC 112 microarray on etiological diagnosis and specific immunotherapy (SIT) prescription. This was compared to the use of conventional diagnoses in pediatric, adolescent, and young adult patients with rhinitis or rhinoconjunctivitis and/or allergic asthma, sensitized to three or more pollen allergens of different botanical species. Methods: A multicenter, prospective, observational study was conducted in patients aged 3–25 years who received care at the Allergology service of 14 hospitals in Catalonia from 2017 to 2020. Allergology diagnosis was established based on the patient’s clinical assessment and the results of the skin prick test and specific immunoglobulin E assays. Subsequently, molecular diagnosis was conducted using ImmunoCAPTM ISAC® 112 to recombinant and/or purified allergen components. Results: A total of 109 patients were included; 35 (32.1%) were pediatric patients and 74 (67.9%) were adolescents or young adults (mean age: 18 years), with 58.0% being females. A change of 51.0% was observed in SIT prescription following molecular etiological diagnosis by means of a multi-parameter microarray. Conclusions: Molecular diagnosis by means of multi-parameter tests increases the accuracy of etiological diagnosis and helps to define an accurate composition of SIT.

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21

Mullenix, Michael C., Steve Wiltshire, Weiping Shao, Gary Kitos, and Barry Schweitzer. "Allergen-specific IgE Detection on Microarrays Using Rolling Circle Amplification: Correlation with in Vitro Assays for Serum IgE." Clinical Chemistry 47, no. 10 (October 1, 2001): 1929. http://dx.doi.org/10.1093/clinchem/47.10.1926.

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22

Shahali, Y., P. Nicaise, A. Brázdová, D. Charpin, E. Scala, A. Mari, J. P. Sutra, S. Chollet-Martin, H. Sēnēchal, and Pascal Poncet. "Complementarity between Microarray and Immunoblot for the Comparative Evaluation of IgE Repertoire of French and Italian Cypress Pollen Allergic Patients." Folia Biologica 60, no. 4 (2014): 192–201. http://dx.doi.org/10.14712/fb2014060040192.

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Cypress pollen represents the primary cause of respiratory allergies in Mediterranean areas. Patients allergic to Cupressus sempervirens pollen (Cups) (CPA) can be discriminated on the basis of the immunoglobulin E (IgE) binding to a basic 14 kDa protein (BP14) or to high-molecular-weight (HMW) glycoproteins only. Specific IgE repertoires of two differentially exposed CPA cohorts, French and Italian, were investigated using an IgE microarray system (some known major allergens from several allergenic sources) and individual IgE immunoblotting (IB) of whole Cups pollen extract separated by SDS-PAGE (all allergens from one allergenic source: cypress pollen). The prevalence of sensitization to BP14 was higher in French (37 %) than in Italian patients (17 %) and major differences were observed in IgE reactivities to lipid transfer proteins (LTPs). Thirty percent of the Italian CPA (4 % in the French group) had specific IgE against the Parietaria pollen LTP, independently of IB subgroups. Regarding peach LTP sensitization, all Pru p 3+ Italian CPA (10 %) were in the HMW+ subgroup, while Pru p 3+ French CPA (20 %) were all included in the BP14+ subgroup. BP14 sensitization is likely a marker of Cups exposure and is, in French CPA, significantly correlated to Pru p 3 sensitization. The IgE immunoblot and microarray are complementary tools that highlight differences in the subtle sensitization profile between groups of patients in comparative studies.

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23

Loffredo, Lucas, and Sergejs Berdnikovs. "Microarray analysis of epithelial-to-mesenchymal transition genes suggests differential epithelial remodeling in human disease vs. mouse models of asthma (MUC1P.912)." Journal of Immunology 194, no. 1_Supplement (May 1, 2015): 64.13. http://dx.doi.org/10.4049/jimmunol.194.supp.64.13.

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Abstract Epithelial-mesenchymal transition (EMT) is the process by which epithelial cells revert to a less differentiated, mesenchymal phenotype; it occurs in maintenance of tissue homeostasis and is exaggerated in disease. EMT has been implicated in human asthma, although it is not clear whether this process is caused by inflammation or rather contributes to it. In allergen-induced murine models of asthma, EMT would be directly caused by inflammation; however, evidence for EMT in such models is lacking. To assess weight-of-evidence and causality for EMT, we contrasted gene expression between mouse models and human asthma samples, using data from 12 gene expression microarrays (GEO, NCBI). PCA analysis surprisingly discriminated human and mouse studies based on behavior of 8 classical EMT genes and 25 epithelial differentiation genes. Increased expression of Cdh2, Twist1, Twist2, Snai2 and Zeb1 supported existence of EMT in human asthma. Interestingly, the same genes did not change or decreased their expression in mouse models. Moreover, inducers of EMT Sox4, Fgf18 and Fgfr4 decreased and suppressor of EMT Runx3 increased expression in mice, consistent with mesenchymal-epithelial transition associated with repair. The fact that even chronic mouse models failed to recapitulate the EMT pattern seen in patients suggests that allergic inflammation alone is not sufficient to trigger persistent epithelial de-differentiation, implicating a missing component in the causation of EMT in asthma.

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24

Mathae, Laura, Itziar Martinez-Gonzalez, and Fumio Takei. "Gene expression profile of allergen-experienced group 2 innate lymphoid cells (HYP2P.323)." Journal of Immunology 194, no. 1_Supplement (May 1, 2015): 53.4. http://dx.doi.org/10.4049/jimmunol.194.supp.53.4.

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Abstract We previously identified group 2 innate lymphoid cells (ILC2s) in mouse lungs. Upon allergen encounter, they produce type 2 cytokines and mediate T cell-independent allergic lung inflammation and also promote Th2 responses. We have recently found that allergen-treatments of mice result in significant increase in the number of ILC2s in the lung and lymph node even after the resolution of inflammation for several weeks. The allergen-experienced ILC2s are more responsive to unrelated allergens than naive ILC2s and rapidly induce allergic lung inflammation. To characterize the allergen-experienced ILC2s, we have performed microarray analysis of ILC2s purified from naïve mice and allergen-treated mice 4 days, 2 weeks, and 4 months after the treatment. The genes that are differentially expressed at different time points include those encoding chemokines, cytokines and their receptors, cell cycle and proliferation related proteins, adhesion molecules, and transcription factors. Gene Set Enrichment Analyses of the ILC2 populations revealed that allergen-experienced ILC2s are enriched for the gene signature of memory T cells. These results suggest that activated ILC2s undergo cell intrinsic changes and acquire memory-like features that are governed by the same set of genes as those responsible for memory T cell differentiation.

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25

Miralles-Lopez, Juan Carlos, Antonio Carbonell-Martínez, Soledad Zamarro-Parra, Cristina Navarro-Garrido, Ana Isabel Escudero-Pastor, Muna Boulaich, Sol Sanromán-Sirvent, Yulia Petryk-Petryk, Maria Dolores Ladrón-de-Guevara, and Virginia Pérez-Fernández. "Clinical and serological characteristics of patients allergic to LTP." Allergologia et Immunopathologia 52, no. 4 (July 1, 2024): 9–14. http://dx.doi.org/10.15586/aei.v52i4.1074.

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Background: Allergy to lipid transfer proteins (LPT) is common in Mediterranean Europe, and it causes severe reactions in patients and affects multiple foods, impairing the quality of life. Objective: This study aimed to describe the clinical and sensitization profile of patients with LTP syndrome and to determine a clinical pattern of severity. Molecular diagnosis is shown in a broad population through microarrays. Material and Methods: This study was performed at the LTP Allergy Consultation of the Reina Sofia Hospital in Murcia, Spain. We analyzed the patients’ characteristics, reactions, cofactors, food implicated, quality of life, skin prick test to food and aeroallergens, and serologic parameters, such as total immunoglobulin E, peach LTP (Pru p 3 IgE) and immunoglobulin G4, and microarray Immuno Solid-phase Allergen Chip (ISAC). We related the severity of the reactions with other variables. Results: We presented a series of 236 patients diagnosed with LTP allergy, 54.66% suffering from anaphylaxis, 36.02% from urticaria angioedema, and 9.32% from oral allergy syndrome. The most frequently implicated food was peach, producing symptoms in 70% of patients, followed by walnut in 55%, peanut in 45%, hazelnut in 44%, and apple in 38% patients. Regarding the food that provoked anaphylaxis, walnut was the most frequent instigator, along with peach, peanut, hazelnut, almond, sunflower seed, and apple. According to the severity of LPT reaction, we did not discover significant differences in gender, age, food group involved, and serologic parameters. We found differences in the presence of cofactors, with 48.84% of cofactors in patients with anaphylaxis, compared to 27.1% in patients without anaphylaxis and in family allergy background (P < 0.0001). Conclusion: In our series of patients, 54% presented anaphylaxis, and the foods that most frequently produced symptoms were peaches, apples, and nuts. Cofactors and family allergy backgrounds were associated with the severity of LPT reaction.

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26

Niespodziana, Katarzyna, Katarina Stenberg-Hammar, Nikolaos G. Papadopoulos, Margarete Focke-Tejkl, Peter Errhalt, Jon R. Konradsen, Cilla Söderhäll, Marianne van Hage, Gunilla Hedlin, and Rudolf Valenta. "Microarray Technology May Reveal the Contribution of Allergen Exposure and Rhinovirus Infections as Possible Triggers for Acute Wheezing Attacks in Preschool Children." Viruses 13, no. 5 (May 15, 2021): 915. http://dx.doi.org/10.3390/v13050915.

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Allergen exposure and rhinovirus (RV) infections are common triggers of acute wheezing exacerbations in early childhood. The identification of such trigger factors is difficult but may have therapeutic implications. Increases of IgE and IgG in sera, were shown against allergens and the N-terminal portion of the VP1 proteins of RV species, respectively, several weeks after allergen exposure or RV infection. Hence, increases in VP1-specific IgG and in allergen-specific IgE may serve as biomarkers for RV infections or allergen exposure. The MeDALL-allergen chip containing comprehensive panels of allergens and the PreDicta RV chip equipped with VP1-derived peptides, representative of three genetic RV species, were used to measure allergen-specific IgE levels and RV-species-specific IgG levels in sera obtained from 120 preschool children at the time of an acute wheezing attack and convalescence. Nearly 20% of the children (22/120) showed specific IgE sensitizations to at least one of the allergen molecules on the MeDALL chip. For 87% of the children, increases in RV-specific IgG could be detected in the follow-up sera. This percentage of RV-specific IgG increases was equal in IgE-positive and -negative children. In 10% of the children, increases or de novo appearances of IgE sensitizations indicative of allergen exposure could be detected. Our results suggest that, in the majority of preschool children, RV infections trigger wheezing attacks, but, in addition, allergen exposure seems to play a role as a trigger factor. RV-induced wheezing attacks occur in IgE-sensitized and non-IgE-sensitized children, indicating that allergic sensitization is not a prerequisite for RV-induced wheeze.

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Figo, Daniele Danella, Priscilla Rios Cordeiro Macedo, Gabriele Gadermaier, Cesar Remuzgo, Fábio Fernandes Morato Castro, Jorge Kalil, Clovis Eduardo Santos Galvão, and Keity Souza Santos. "IgE and IgG4 Epitopes of Dermatophagoides and Blomia Allergens before and after Sublingual Immunotherapy." International Journal of Molecular Sciences 24, no. 4 (February 20, 2023): 4173. http://dx.doi.org/10.3390/ijms24044173.

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Sublingual immunotherapy (SLIT) is used worldwide to treat house dust mites (HDM) allergy. Epitope specific immunotherapy with peptide vaccines is used far less, but it is of great interest in the treatment of allergic reactions, as it precludes the drawbacks of allergen extracts. The ideal peptide candidates would bind to IgG, blocking IgE-binding. To better elucidate IgE and IgG4 epitope profiles during SLIT, sequences of main allergens, Der p 1, 2, 5, 7, 10, 23 and Blo t 5, 6, 12, 13, were included in a 15-mer peptide microarray and tested against pooled sera from 10 patients pre- and post-1-year SLIT. All allergens were recognized to some extent by at least one antibody isotype and peptide diversity was higher post-1-year SLIT for both antibodies. IgE recognition diversity varied among allergens and timepoints without a clear tendency. Der p 10, a minor allergen in temperate regions, was the molecule with more IgE-peptides and might be a major allergen in populations highly exposed to helminths and cockroaches, such as Brazil. SLIT-induced IgG4 epitopes were directed against several, but not all, IgE-binding regions. We selected a set of peptides that recognized only IgG4 or were able to induce increased ratios of IgG4:IgE after one year of treatment and might be potential targets for vaccines.

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Szameit, Sandra, Klemens Vierlinger, Letizia Farmer, Helga Tuschl, and Christa Noehammer. "Microarray-Based In Vitro Test System for the Discrimination of Contact Allergens and Irritants: Identification of Potential Marker Genes." Clinical Chemistry 54, no. 3 (March 1, 2008): 525–33. http://dx.doi.org/10.1373/clinchem.2007.097386.

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Abstract Background: Animal tests have been used to characterize the potential of chemicals to produce allergic contact dermatitis, but this approach is increasingly a matter of public and political concern. Our aim was to develop and validate an alternative in vitro test that can identify contact allergens. Methods: We developed a targeted microarray containing oligonucleotide probes for 66 immune-relevant genes and analyzed gene expression in monocyte-derived dendritic cells (Mo-DCs) treated with 1 irritant (SDS) and 2 prominent contact allergens, nickel and Bandrowski’s base (BB), which is the oxidation product of the most important hair dye allergen, p-phenylenediamine. Results: Comparing RNA amounts in chemical-treated and solvent-treated cells, we identified significant changes in the expression of 21 genes and 10 genes after exposure of immature DCs (iDCs) to nickel and BB, respectively, but not after exposure to SDS. Eight genes were differentially expressed after application of both nickel and BB. Real-time PCR was used to confirm the results for selected genes. Conclusion: We propose a microarray-based in vitro test that might allow the identification of contact allergens. Independently from donor variability, several immune-relevant genes were up- or downregulated after the application of the tested sensitizers to iDCs, therefore presenting potential marker genes. While reducing the number of laboratory animals used, this test would also enable reliable analysis of chemicals using a human system.

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Orga-Dumitriu, Dan, Dana M. Harris, and Corina Porr. "Anaphylactic Shock Caused by Eating Buckwheat." Journal of Clinical Medicine 13, no. 17 (September 4, 2024): 5243. http://dx.doi.org/10.3390/jcm13175243.

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Background: Urticaria is a common disease with a marked influence on quality of life. The key cell involved is the mast cell, which can be activated by a vast variety of stimuli, and the major mediator is histamine. Allergic urticaria is a disorder with a large variety of causes: food, drugs, insect venom, skin contact with allergens, and physical exercise. Buckwheat consumption has increased in European countries and the USA because it is gluten-free. It can trigger anaphylactic shock if ingested, inhaled, or handled with the hands. Five common buckwheat allergens named Fag e1 to 5 (Fag e1, 2, and 3 are considered the major allergens) and two tartary buckwheat allergens named Fag t1 and Fag t2 have been described. Method: We present the case of a patient who experienced two anaphylactic shocks and in whom the etiological factor was buckwheat. The patient presented to the Allergology department for the evaluation of two episodes of severe allergic reactions that required emergency therapy, episodes that involved the loss of consciousness and were of major severity. At each anaphylactic shock, an ambulance was requested, and emergency therapy was administered, leading to the patient’s recovery within a few hours. Diagnosis: Since each episode occurred a few minutes after eating, the diagnosis was established based on a detailed anamnesis and prick skin tests, followed by specific IgE dosages. Other foods consumed by the patient, assessed by prick skin testing and specific IgE dosages of suspected foods, were excluded as the etiological cause. Increased levels of buckwheat-specific immunoglobulin E were highlighted, thus identifying the etiological agent. The treatment of anaphylactic shock was performed urgently by the ambulance crew with adrenaline, infusion solutions, cortisone preparations, and antihistamines. Result: Following the treatment that was initiated, there was a partial remission of the lesions after a few hours. Conclusions: Buckwheat allergy is rare, but it produces symptoms that affect the skin, gastrointestinal tract, and respiratory tract, as well as anaphylaxis. In a professional environment, it can trigger allergic rhinitis, asthma, and hives. Although buckwheat allergens have been described, their clinical relevance has only been studied in a small number cases. In current practice, the only commercially available allergen is Beech e2 per the ImmunoCAP ISAC microarray. Diagnosis can be difficult in clinical practice. This reported case suggests the need for a thorough anamnesis, since buckwheat is consumed as a hidden allergen, and in Europe, it is not necessary to label foods containing this allergen.

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Joshi, Amit A., Mark W. Peczuh, Challa V. Kumar, and James F. Rusling. "Ultrasensitive carbohydrate-peptide SPR imaging microarray for diagnosing IgE mediated peanut allergy." Analyst 139, no. 22 (2014): 5728–33. http://dx.doi.org/10.1039/c4an01544d.

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The first peptide-carbohydrate SPR imaging immunoarray aimed at diagnosing severity of peanut allergies is reported. The SPRi chip features a peptide epitope of peanut protein allergen Ara h2, a β-xylosyl glycoside, and anti-IgE antibody to achieve epitope-specific detection of human allergen IgEs at sub-attomol levels.

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Birras, Jasmin, Samuel J. White, Sigridur Jonsdottir, Ella N. Novotny, Anja Ziegler, A. Douglas Wilson, Rebecka Frey, Sigurbjörg Torsteinsdottir, Marcos Alcocer, and Eliane Marti. "First clinical expression of equine insect bite hypersensitivity is associated with co-sensitization to multiple Culicoides allergens." PLOS ONE 16, no. 11 (November 15, 2021): e0257819. http://dx.doi.org/10.1371/journal.pone.0257819.

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Background Insect bite hypersensitivity (IBH) is an IgE-mediated allergic dermatitis in horses incited by salivary allergens from Culicoides spp. IBH does not occur in Iceland, as the causative agents are absent, however a high prevalence is seen in horses exported to Culicoides-rich environments. Aims To study the natural course of sensitization to Culicoides allergens and identify the primary sensitizing allergen(s) in horses exported from Iceland utilizing a comprehensive panel of Culicoides recombinant (r-) allergens. Method IgE microarray profiling to 27 Culicoides r-allergens was conducted on 110 serological samples from horses imported to Switzerland from Iceland that subsequently developed IBH or remained healthy. Furthermore, a longitudinal study of 31 IBH horses determined IgE profiles the summer preceding first clinical signs of IBH (TIBH-1), the summer of first clinical signs (TIBH) and the following summer (TIBH+1). In a group of Icelandic horses residing in Sweden, effects of origin (born in Iceland or Sweden) and duration of IBH (<4 years, 4–7 years, >7 years) on Culicoides-specific IgE was evaluated. Sero-positivity rates and IgE levels were compared. Results At TIBH, horses were sensitized to a median of 11 r-allergens (range = 0–21), of which nine were major allergens. This was significantly higher than TIBH-1 (3, 0–16), as well as the healthy (1, 0–14) group. There was no significant increase between TIBH and TIBH+1(12, 0–23). IBH-affected horses exported from Iceland had a significantly higher degree of sensitization than those born in Europe, while duration of IBH did not significantly affect degree of sensitization. Conclusion Significant sensitization is only detected in serum the year of first clinical signs of IBH. Horses become sensitized simultaneously to multiple Culicoides r-allergens, indicating that IgE-reactivity is due to co-sensitization rather than cross-reactivity between Culicoides allergens. Nine major first sensitizing r-allergens have been identified, which could be used for preventive allergen immunotherapy.

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Halim, Timotheus, Ann Sun, and Fumio Takei. "Lung natural helper cells are an important source of innate TH2 cytokines (158.2)." Journal of Immunology 186, no. 1_Supplement (April 1, 2011): 158.2. http://dx.doi.org/10.4049/jimmunol.186.supp.158.2.

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Abstract Recently, novel innate TH2 cytokine producing lymphocytes termed nuocytes or natural helper cells have been found in the gut-associated mucosa tissues. Similar TH2 cytokine producing innate cells have also been detected in other organs, including the lung, but their functional significance is not well understood. We have identified TH2 cytokine producing innate lymphocytes, termed lung natural helper (NH) cells in B6 mouse lungs and elucidated their role in lung eosinophilia and asthma. Lung NH cells are Lineage-negative but express of Sca-1, c-Kit, CD127 and CD25. Lung NH cell gene expression microarrays indicate that they likely constitute a novel lymphoid lineage. When purified and stimulated in vitro, they produce large amounts of IL3, 4, 5, 13, and IL17A. They also rapidly produce IL5 and IL13 in lung explant cultures stimulated with IL25, an alarmin released by stressed lung epithelial cells. Lung NH cells are present in Rag-/- mice that lack T and B cells but absent in Rag-/-cγ-/- mice. Intranasal administration of protease allergen papain rapidly induces eosinophil infiltration and mucus hypersecretion, which are hallmark asthma symptoms, in the lung of B6 and Rag-/- mice but not in Rag-/-cγ-/- mice. Papain treatment of lung explants also induces IL13 production by lung NH cells. Thus, lung NH cells are unique lymphocytes of the innate immune system and an important source of T cell-independent TH2 cytokines. They likely play an important role in initiating asthma.

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Jahn-Schmid, B., C. Harwanegg, R. Hiller, B. Bohle, C. Ebner, O. Scheiner, and M. W. Mueller. "Allergen microarray: comparison of microarray using recombinant allergens with conventional diagnostic methods to detect allergen-specific serum immunoglobulin E." Clinical & Experimental Allergy 33, no. 10 (October 2003): 1443–49. http://dx.doi.org/10.1046/j.1365-2222.2003.01784.x.

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Kalli, Marina, Andrew Blok, Marcos Alcocer, and Franco Falcone. "Protein microarrays: The future of allergy diagnosis? Optimization of coating and printing of allergen arrays used with fluorescent humanised basophil reporter cell line NFAT-DsRed RBL." World Allergy Organization Journal 13, no. 8 (August 2020): 100407. http://dx.doi.org/10.1016/j.waojou.2020.100407.

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Hsiao, Yun-Hsia, Charles Chen, and Ton Willemse. "Allergen Sensitization Patterns of Allergic Dogs: IgE-microarray Analysis." Thai Journal of Veterinary Medicine 46, no. 2 (June 1, 2016): 235–42. http://dx.doi.org/10.56808/2985-1130.2731.

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Hochwallner, H., U. Schulmeister, I. Swoboda, N. Balic, B. Geller, M. Nystrand, A. Härlin, et al. "Microarray and allergenic activity assessment of milk allergens." Clinical & Experimental Allergy 40, no. 12 (September 22, 2010): 1809–18. http://dx.doi.org/10.1111/j.1365-2222.2010.03602.x.

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Preda, Mariana, Florin-Dan Popescu, Emilia Vassilopoulou, and Sylwia Smolinska. "Allergenic Biomarkers in the Molecular Diagnosis of IgE-Mediated Wheat Allergy." International Journal of Molecular Sciences 25, no. 15 (July 27, 2024): 8210. http://dx.doi.org/10.3390/ijms25158210.

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IgE-mediated wheat allergy can take on various forms, including childhood food allergy to wheat, wheat-dependent exercise-induced anaphylaxis in young adults, baker’s respiratory allergy/asthma in workers exposed to wheat flour inhalation, and contact urticaria that is caused by hydrolyzed wheat proteins in some cosmetics, and that is sometimes associated with a food allergy. Singleplex and multiplex immunoassays detect specific IgE antibodies to wheat allergenic molecular biomarkers such as omega-5 gliadin Tri a 19, lipid transfer protein Tri a 14, and alpha-amylase inhibitors. The fluorescence enzyme immunoassay with capsulated cellulose polymer solid-phase coupled allergens is a commonly used singleplex assay. Multiplex methods include the ELISA-based macroarray immunoassay using nano-bead technology and a microarray immunoassay on polymer-coated slides. Another promising diagnostic tool is the basophil activation test performed with omega-5 gliadin and other wheat protein types. Detailed comprehension of the structural and immunological features of the numerous wheat allergens significant in clinical settings is imperative for advancing diagnostic biomarkers for IgE-mediated wheat allergies.

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Mari, Adriano, Claudia Alessandri, Maria Livia Bernardi, Rosetta Ferrara, Enrico Scala, and Danila Zennaro. "Microarrayed Allergen Molecules for the Diagnosis of Allergic Diseases." Current Allergy and Asthma Reports 10, no. 5 (July 2, 2010): 357–64. http://dx.doi.org/10.1007/s11882-010-0132-0.

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Rudokas, Vytautas, Laimis Silimavicius, Indre Kucinskaite-Kodze, Aiste Sliziene, Milda Pleckaityte, and Aurelija Zvirbliene. "Novel monoclonal antibodies against house dust mite allergen Der p 21 and their application to analyze allergen extracts." PeerJ 12 (April 18, 2024): e17233. http://dx.doi.org/10.7717/peerj.17233.

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Background Allergen extracts and recombinant allergens are used in allergy diagnostics and immunotherapy. Since allergen extracts from different manufacturers lack proper standardization regarding their composition, monoclonal antibodies (MAbs) against specific allergen components can be used for their identification and quantification in allergen extracts. This study aimed to generate MAbs against allergen Der p 21 of Dermatophagoides pteronyssinus for the analysis of allergen extracts. Methods Recombinant Der p 21 was expressed in E. coli and purified using affinity chromatography. MAbs against Der p 21 were generated using hybridoma technology. House dust mite (HDM) allergen extracts were analyzed using the newly developed sandwich enzyme-linked immunosorbent assay, Western blotting and microarray immunoassay. Results MAbs raised against recombinant Der p 21 were characterized in detail and proven to be reactive with natural Der p 21. Highly specific sandwich enzyme-linked immunosorbent assay for the quantification of Der p 21 was developed and optimized. The allergen was detected and its concentration was determined in only three of six analyzed HDM allergen extracts from different manufacturers. Conclusion HDM analysis by MAb-based immunoassays shows their differences in allergen composition. The results demonstrate the importance of allergen-specific MAbs as a tool for the characterization of allergen extracts and the need for their appropriate standardization before their use for allergy diagnostics or immunotherapy.

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Moreira, Priscila Ferreira de Sousa, Katharina Gangl, Francisco de Assis Machado Vieira, Leandro Hideki Ynoue, Birgit Linhart, Sabine Flicker, Helmut Fiebig, et al. "Allergen Microarray Indicates Pooideae Sensitization in Brazilian Grass Pollen Allergic Patients." PLOS ONE 10, no. 6 (June 11, 2015): e0128402. http://dx.doi.org/10.1371/journal.pone.0128402.

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Fortino, Vittorio, Lukas Wisgrill, Paulina Werner, Sari Suomela, Nina Linder, Erja Jalonen, Alina Suomalainen, et al. "Machine-learning–driven biomarker discovery for the discrimination between allergic and irritant contact dermatitis." Proceedings of the National Academy of Sciences 117, no. 52 (December 14, 2020): 33474–85. http://dx.doi.org/10.1073/pnas.2009192117.

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Contact dermatitis tremendously impacts the quality of life of suffering patients. Currently, diagnostic regimes rely on allergy testing, exposure specification, and follow-up visits; however, distinguishing the clinical phenotype of irritant and allergic contact dermatitis remains challenging. Employing integrative transcriptomic analysis and machine-learning approaches, we aimed to decipher disease-related signature genes to find suitable sets of biomarkers. A total of 89 positive patch-test reaction biopsies against four contact allergens and two irritants were analyzed via microarray. Coexpression network analysis and Random Forest classification were used to discover potential biomarkers and selected biomarker models were validated in an independent patient group. Differential gene-expression analysis identified major gene-expression changes depending on the stimulus. Random Forest classification identified CD47, BATF, FASLG, RGS16, SYNPO, SELE, PTPN7, WARS, PRC1, EXO1, RRM2, PBK, RAD54L, KIFC1, SPC25, PKMYT, HISTH1A, TPX2, DLGAP5, TPX2, CH25H, and IL37 as potential biomarkers to distinguish allergic and irritant contact dermatitis in human skin. Validation experiments and prediction performances on external testing datasets demonstrated potential applicability of the identified biomarker models in the clinic. Capitalizing on this knowledge, novel diagnostic tools can be developed to guide clinical diagnosis of contact allergies.

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Doyen, Virginie, Carine Truyens, Hoa Nhu Thi, Hiep Tran Thi Mong, Thanh Le Chi, Frederic De Blay, Phuong Thi Ngoe Huynh, Olivier Michel, and Francis Corazza. "Helminth infection induces non-functional sensitization to house dust mites." PLOS ONE 16, no. 7 (July 1, 2021): e0253887. http://dx.doi.org/10.1371/journal.pone.0253887.

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Background IgE characterizes the humoral response of allergic sensitization but less is known about what modulates its function and why some patients present clinical symptoms for a given IgE level and others do not. An IgE response also occurs during helminth diseases, independently of allergic symptoms. This response could be a model of non-functional IgE. Objective To study the IgE response against environmental allergens induced during natural helminth infection. Methods In 28 non allergic subjects from the periphery of Ho Chi Minh city with (H+, n = 18) and without helminth infection (H-, n = 10), we measured IgE and IgG4 against several components of Dermatophagoïdes pteronyssinus (Dpt) and Ascaris (a marker of immunization against nematodes), and determined the IgE component sensitization profile using microarray ISAC biochips. The functional ability of IgE to induce degranulation of cultured mast cells was evaluated in the presence of Dpt. Results Non allergic H+ subjects exhibited higher levels of IgE against Dpt compared to H- subjects. Dpt IgE were not functional in vitro and did not recognize usual Dpt major allergens. IgE recognized other component allergens that belong to different protein families, and most were glycosylated. Depletion of IgE recognizing carbohydrate cross-reactive determinant (CCD) did not induce a reduction in Dpt IgE. The Dpt IgG4 were not significantly different. Conclusion Helminth infections induced IgE against allergens such as Dpt and molecular components that belong to different sources as well as against CCD (such as β-1,2-xylose and/or ⍺-1,3-fucose substituted N-glycans). Dpt IgE were not able to induce degranulation of mast cells and were not explained by sensitization to usual major allergens or N-glycans.

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Sena-Torralba, Amadeo, Javier Gabaldón-Atienza, Aitor Cubells-Gómez, Patricia Casino, Ángel Maquieira, and Sergi Morais. "Lateral Flow Microimmunoassay (LFµIA) for the Reliable Quantification of Allergen Traces in Food Consumables." Biosensors 12, no. 11 (November 7, 2022): 980. http://dx.doi.org/10.3390/bios12110980.

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Quality assurance and food safety are of great concern within the food industry because of unknown quantities of allergens often present in food. Therefore, there is an ongoing need to develop rapid, sensitive, and easy to use methods that serve as an alternative to mass spectrometry and enzyme-linked immunosorbent assay (ELISA) for monitoring food safety. Lateral flow immunoassay is one of the most used point-of-need devices for clinical, environmental, and food safety applications. Compared to traditional methods, it appears to be a simple and fast alternative for detecting food allergens. However, its reliability is frequently questioned due to the lack of quantitative information. In this study, a lateral flow microimmunoassay (LFµIA) is presented that integrates up to 36 spots in microarray format in a single strip, providing semi-quantitative information about the level of allergens, positive and negative controls, internal calibration, and hook effect. The LFµIA has been evaluated for the on-site simultaneous and reliable quantification of almond and peanut allergens as a proof of concept, demonstrating high sensitivity (185 and 229 µg/kg, respectively), selectivity (77%), and accuracy (RSD 5–25%) when analyzing commercial allergen-suspicious food consumables.

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polikepahad, sumanth, Jad El-Daye, Arash Naghavi, Jonathan Miller, Preethi Gunaratne, and David Brian Corry. "Lung RNA profiling suggests an essential role for micro RNAs in regulating allergic lung disease (91.9)." Journal of Immunology 178, no. 1_Supplement (April 1, 2007): S161. http://dx.doi.org/10.4049/jimmunol.178.supp.91.9.

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Abstract MicroRNAs (miRNAs) are small non-coding RNAs, which negatively regulate gene expression by translational inhibition of target mRNAs. In vertebrates, miRNAs are produced in most organs, including lungs. In the current study, we hypothesized that the expression of miRNAs would correlate inversely with expression of critically important target genes involved in the pathogenesis of allergic lung disease. After intranasal challenge of mice with a potent allergen derived from Aspergillus oryzae on alternating days for 2 weeks, lungs were isolated, high quality RNA was extracted and subjected to microarray analysis in comparison to naïve lungs obtained in parallel. In allergic lungs, ~ 30 known miRNAs were suppressed and ~100 were induced, including many new miRNA candidates uncovered through genome-wide predictions. Bioinformatics analysis revealed that miRNA 199a*, which was suppressed, and miR-144 which was induced in allergic lungs, are candidate regulators of STAT-6, GATA-3 and costimulatory factors, all of which are critical mediators of allergic lung disease. These studies suggest the existence of many new miRNAs and pivotal role for miRNA 199a* in controlling the expression of allergic lung disease.

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Sano, Akiyo, Akiko Yagami, Kayoko Suzuki, Yohei Iwata, Tsukane Kobayashi, Masaru Arima, Yasuto Kondo, Tetsushi Yoshikawa, and Kayoko Matsunaga. "Two Cases of Occupational Contact Urticaria Caused by Percutaneous Sensitization to Parvalbumin." Case Reports in Dermatology 7, no. 2 (August 29, 2015): 227–32. http://dx.doi.org/10.1159/000439080.

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Background: In recent years, it has been proposed that the primary mechanism for the development of food allergies is percutaneous sensitization. Since 2010, in Japan, the number of immediate-type wheat allergy due to hydrolyzed wheat protein has dramatically increased among those who have been using soap containing hydrolyzed wheat. This incidence supports the hypothesis that food allergens arise through percutaneous sensitization. Clinical Summary: A 25-year-old man (case 1) and an 18-year-old girl (case 2) with atopic dermatitis visited our Department because of food allergy and hand eczema. After starting their work with fish, severe itchy eczema appeared on their hands. They subsequently started to experience oral allergic symptoms, intraoral itchiness and dyspnea after eating fish. Specific IgE antibodies were detected for many fishes, and skin prick tests showed positive reactions for a variety of fishes in both cases. Furthermore, the fluorescence intensities of specific IgE antibodies against parvalbumin from various types of fish in microarray immunoassay analysis showed positive reactions. We diagnosed them as contact urticaria caused by percutaneous sensitization to parvalbumin through job-related physical contact with fish. Conclusion: The patients' histories and findings indicate the possibility of percutaneous sensitization through occupational exposure to parvalbumin, leading to food allergy.

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Lupinek, Christian, Eva Wollmann, and Rudolf Valenta. "Monitoring Allergen Immunotherapy Effects by Microarray." Current Treatment Options in Allergy 3, no. 2 (April 20, 2016): 189–203. http://dx.doi.org/10.1007/s40521-016-0084-2.

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Mathae, Laura, Itziar Martinez-Gonzalez, and Fumio Takei. "Characterization of Allergen Experienced Group 2 Innate Lymphoid Cells: where do they come from, where do they go?" Journal of Immunology 196, no. 1_Supplement (May 1, 2016): 191.8. http://dx.doi.org/10.4049/jimmunol.196.supp.191.8.

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Abstract We identified group 2 innate lymphoid cells (ILC2s) in mouse lungs. Upon exposure to the protease allergen papain, lung ILC2s produced large amounts of IL-5 and IL-13 and induced eosinophilia and mucus hyperproduction. Intranasal injections of recombinant IL-33 also stimulated lung ILC2s. ILC2s that were previously stimulated by an allergen persisted long after the initial inflammation was resolved and responded to allergen challenge more vigorously than naïve ILC2s. Microarray analyses of the gene expression profiles of ILC2s isolated from naïve and allergen-treated mice showed that allergen-experienced ILC2s have a similar gene profile to that of memory T cells. To further study the process of “memory ILC2” generation, we analyzed the expression of cell surface molecules on ILC2s from untreated mice and those after intranasal IL-33 injections at various time points, which showed differential expression of CD27, CD103, CD41 and the IL-25 receptor (IL-25R). Memory ILC2s expressed higher level of IL-25R than naïve ILC2s, and intranasal injection of IL-25 stimulated memory, but not naïve, lung ILC2s. Intranasal IL-33 injections also increased ILC2 numbers in the lung draining mediastinal lymph node (mLN), peripheral blood, spleen and liver but not in the intestine or skin. Phenotype analysis of those ILC2s suggested that activated lung ILC2s migrated into mLN and the blood, and some settled in the liver. The results indicated that IL-25-responsive memory ILC2s are generated and persist in the lung as well as in mLN while the role of the latter in allergic lung inflammation induced by intranasal allergen challenge in sensitized mice is yet to be determined.

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Melioli, Giovanni, Enrico Compalati, Sergio Bonini, and Giorgio W. Canonica. "The added value of allergen microarray technique to the management of poly-sensitized allergic patients." Current Opinion in Allergy and Clinical Immunology 12, no. 4 (August 2012): 434–39. http://dx.doi.org/10.1097/aci.0b013e32835535b8.

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Kutateladze, Tamara, Kakha Bitskinashvili, Nelly Sapojnikova, Tamar Kartvelishvili, Nino Asatiani, Boris Vishnepolsky, and Nelly Datukishvili. "Development of Multiplex PCR Coupled DNA Chip Technology for Assessment of Endogenous and Exogenous Allergens in GM Soybean." Biosensors 11, no. 12 (November 26, 2021): 481. http://dx.doi.org/10.3390/bios11120481.

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Allergenicity assessment of transgenic plants and foods is important for food safety, labeling regulations, and health protection. The aim of this study was to develop an effective multi-allergen diagnostic approach for transgenic soybean assessment. For this purpose, multiplex polymerase chain reaction (PCR) coupled with DNA chip technology was employed. The study was focused on the herbicide-resistant Roundup Ready soya (RRS) using a set of certified reference materials consisting of 0, 0.1%, 0.5%, and 10% RRS. Technically, the procedure included design of PCR primers and probes; genomic DNA extraction; development of uniplex and multiplex PCR systems; DNA analysis by agarose gel electrophoresis; microarray development, hybridization, and scanning. The use of the asymmetric multiplex PCR method is shown to be very efficient for DNA hybridization with biochip probes. We demonstrate that newly developed fourplex PCR methods coupled with DNA-biochips enable simultaneous identification of three major endogenous allergens, namely, Gly m Bd 28K, Gly m Bd 30K, and lectin, as well as exogenous 5-enolppyruvyl shikimate-phosphate synthase (epsps) expressed in herbicide-resistant roundup ready GMOs. The approach developed in this study can be used for accurate, cheap, and fast testing of food allergens.

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Incorvaia, Cristoforo, Marina Mauro, Erminia Ridolo, Eleni Makrì, Marcello Montagni, and Giorgio Ciprandi. "A Pitfall to Avoid When Using an Allergen Microarray: The Incidental Detection of IgE to Unexpected Allergens." Journal of Allergy and Clinical Immunology: In Practice 3, no. 6 (November 2015): 879–82. http://dx.doi.org/10.1016/j.jaip.2014.09.020.

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