Allergenic Components Research Articles

Extended IgE profile of shrimp-sensitized patients based on Multiplex Examination ALEX2 - Allergy Explorer.

The key to the correct diagnosis of shrimp allergy is a qualification to the most efficient diagnostic method and later interpretation of the result. To achieve this, it is necessary to apply a diagnostic strategy relevant to each patient's clinical situation and approach every case individually. In this study the allergen profile of shrimp-sensitized patients was analysed using ALEX2 Allergy Explorer. This study includes 50 adult patients with positive prick-by-prick tests with tiger shrimp bought from the local eco-market and an elevated concentration of IgE specific to the shrimp allergen extract (ImmunoCap). A total of 35 patients with negative skin prick tests with shrimp and not detectable sIgE shrimp in ImmunoCap were included in the control group. All patients had ALEX2 Allergy Explorer microarray test. In the shrimp-sensitized group, 22 patients were sensitized to at least one allergen component of Penaeus monodon, 20 patients were sensitized to crab, and 20 were sensitized to lobster. Only 15 (30%) patients were sensitized to the Northern prawn (Pandalus borealis) allergen extract in ALEX2 and only 12 (24%) to Shrimp mix (Litopenaeus setiferus, Farfantepenaeus aztecus, Farfantepenaeus dourarum). Sensitization to shrimp tropomyosin in the research group was present only in 34% of cases. There may be other shrimp allergen components, not available in ALEX2, which are responsible for shrimp sensitization.

Determination of the mycobiome in the lower respiratory tract of horses with equine asthma

Fungal particles are important allergenic components involved in the development of equine asthma. The aim of this study was to evaluate the mycobiome composition of the lower respiratory tract in asthmatic horses using fungal culture, quantitative multiplex real-time PCR analysis (FungiMultiPlex) and Next-Generation Sequencing approach. Bronchoalveolar lavage fluid (BALF) samples obtained from 45 client-owned horses diagnosed with equine asthma were analysed by fungal culture (19 samples), FungiMultiPlex (34 samples), and Next-Generation Sequencing (14 samples). The fungal culture was positive in 11/19 (58%) cases, and FungiMultiPlex was positive in 19/34 (56%) cases. No fungal PCR product was detected by Next-Generation Sequencing analysis. Fungal culture and FungiMultiPlex methods were performed simultaneously on eight horses only. Association of results of these methods was calculated using Phi coefficient (φ= 0.333), and concordance between the methods was not confirmed (P = 0.420). The results of this study suggest that the fungal culture and quantitative multiplex real-time PCR might be considered diagnostically useful to assess the presence of fungi in BALF in a semiquantitative and quantitative manner, respectively. The Next-Generation Sequencing method seems to be less diagnostically suitable due to technical obstacles pertinent to both the low concentration of microbial agents in the rather diluted BALF samples, and, also, due to the relatively high environmental background contamination of the collected material. Based on our data, we advocate the use of the combination of quantitative multiplex real-time PCR and fungal culture in a routine clinical diagnostic setting.

Molecular peanut sensitization patterns in Lithuanian children with suspected allergic symptoms.

There are no in-depth studies describing the peanut sensitization molecular patterns in Lithuanian children. To investigate the age-related patterns of molecular peanut sensitization profiles in Lithuanian children with suspected allergic symptoms. We performed a retrospective analysis of peanut sensitization profiles in 576 Lithuanian children with possible allergic symptoms. Patient data were categorized according to age groups: 0-2, 3-6, 7-12, and 13-18 years. Specific immunoglobulin E levels to peanut molecular components: Ara h 1, Ara h 2, Ara h 3, Ara h 6, Ara h 8, Ara h 9, Ara h 15, and birch major allergen component Bet v 1 were analyzed. Sensitization to at least one peanut protein was observed in 148 (25.7%) children. In children aged ≤ 2 years, most children were sensitized to Ara h 1 - 11 (12.1%). In children aged from 3 to 6 years, the sensitization to Ara h 1 remained the most prevalent in 40 (16.6%) children. The most prevalent sensitization in children aged from 7 to 12 years was to Ara h 8 in 39 (24.5%) of them. In children aged ≥ 13 years, Ara h 8 remained the most prevalent sensitizer in 21 (24.7%) of them. One-fourth of children with suspected allergic symptoms are sensitized to at least one peanut molecular component in Lithuania. Infants and preschool children are most commonly sensitized to seed storage proteins, while school-age children to Ara h 8 allergen.

Микологические факторы риска развития респираторных аллергозов тяжелого течения у населения, проживающего на юге России

Aim: to determine the concentration of mold spores and causative allergens of pollen-producing grasses in the cities of Southern Russia and to assess the prevalence of multisensitization in patients with severe allergic respiratory diseases, taking into consideration mold specific IgE results. Patients and Methods: from 2018 to 2020, concentrations of airborne allergens were evaluated in Stavropol, Krasnodar and Rostov-on-Don according to international techniques, using Burkard and Lanzoni volumetric spore and pollen traps. The levels of non-bacterial allergen- specific IgE were evaluated by allergen component testing in 143 patients (69 children and 74 adults) with allergic rhinitis and atopic asthma. Results: the results of aeropalynology studies demonstrated a high concentration of mold spores in the atmospheric air of all the studied cities co-occurring with seasonal increases in pollen allergen concentrations in the atmospheric air. The South of Russia is characterized by the year-round presence of mold allergens with sporulation peaks in July, August and September. The peak utilization of healthcare services coincided with the maximal concentration of plant pollen and mold spores. The results of comprehensive allergy diagnostic testing suggest that there is a predominance of patients with multisensitization (71%), including sensitization to pollen and mold spore allergens. As a result of the overlapping of the peaks of spreading airborne pollen allergens and mold sporulation, a frequent use of short-acting emergency β2-agonists and more calls for ambulance service are reported. Conclusion: the predominance of patients with multisensitization leads to the development of severe allergic respiratory diseases. Measures of timely management and prevention are needed for such patients. A daily monitoring of sporulation levels and pollen allergens in the air may help to avoid contacts with high concentrations of causative allergens and to implement the necessary preventive measures. The annual aeropalynology monitoring will provide an opportunity to predict allergen concentrations in the atmospheric air during the current year and to develop a statistically significant monitoring model in the future. KEYWORDS: molds, ragweed, aeromycology, aeropalynology, allergens, allergen component testing. FOR CITATION: Ukhanova O.P., Bogomolova E.V., Budnikov P.V. et al. Mycotic risk factors for the development of severe respiratory allergosis in the population of southern Russia. Russian Medical Inquiry. 2023;7(2):65–74 (in Russ.). DOI: 10.32364/2587-6821-2023-7-2-65-74.

Identification of Pollen and Food Sensitization Prevalence in Children with Different Atopic Phenotype Variants (Intermediate Study Results)

Background. Identification of the prevalence of pollen and food allergen molecules sensitization in children (considering regional characteristics) is relevant and crucial for personalized preventive and treatment measures development. Objective. The aim of the study is — study the prevalence of pollen and food allergen sensitization in children with different atopic phenotype variants. Methods. This cross-sectional study included 80 children aged from 4 months to 17 years 8 months with symptoms of allergic rhinitis, food allergy, atopic dermatitis living in the Moscow metropolitan area. All children were sensitized to 9 extracts of domestic, pollen and food allergens via the ImmunoCAP method, and molecular allergy diagnosis was carried out via the ImmunoCAP ISAC technology (n = 79). One patient with symptoms of allergic rhinitis during spring without any manifestations of cross-reactive food allergy received an unreadable result of ImmunoCAP ISAC. Results. The most common sensitization among all patients was to the birch pollen allergen extract — 65%, herd grass — 46%, common mugwort — 32%. The following sensitization to major allergens was most often detected according to molecular diagnosis: birch pollen Bet v 1 — 67%, herd grass Phl p 1 — 33% and Phl p 4 — 24%, and even less mugwort Art v 1 — 11%. The lowest sensitization was to the allergy component Amb a 1 of ambrosia — 8% of cases. Sensitization to wormwood pollen extract was revealed 2.5 times more often than to major components of this allergen. Spring allergic rhinitis was diagnosed in 45 children, and 27 of them had cross-reactive food allergy. Children with allergic rhinitis during the spring period (both with or without complaints on cross-reactive food allergy) most commonly had sensitization to the allergens components of hazelnut Cor a 1.0401 and apple Mal d 1, and least often to the component of the celery allergen Api g 1. Conclusion. Birch pollen is the most common primary allergen in children of the Moscow metropolitan area with allergy. Patients with spring allergic rhinitis despite the presence of cross-reactive food allergy usually have sensitization to the components of hazelnut and apple allergens.

Identification and characterization of natural PR-1 protein as major allergen from Humulus japonicus pollen

BackgroundThe Humulus japonicus pollen is one of the most common allergenic pollens in China. However, little is unveiled regarding the allergenic components in Humulus japonicus pollen. Our study aimed to purify and identify the pathogenesis-related 1 (PR-1) protein from Humulus japonicus pollen, and to characterize the molecular and immunochemical properties of this novel allergen. MethodsThe natural PR-1 protein (named as Hum j PR-1) was purified from Humulus japonicus pollen extracts with a combined strategy of chromatography, and identified by mass spectrometry. The coding sequence of Hum j PR-1 was confirmed by cDNA cloning. The recombinant Hum j PR-1 was expressed and purified from Escherichia coli. The allergenicity was assessed by immunoblot, enzyme-linked immunosorbent assay (ELISA), inhibition ELISA, and basophil activation test using Humulus japonicus allergic patients’ whole blood. The physicochemical properties and 3-dimensional structure of it were comprehensively characterized by in silico methods. ResultsThe allergenicity analysis revealed that 76.6 % (23/30) of the Humulus japonicus pollen allergic patients displayed specific IgE recognition of the natural Hum j PR-1. The cDNA sequence of Hum j PR-1 had a 516-bp open reading frame encoding 171 amino acids. Physicochemical analysis indicated that Hum j PR-1 was a stable and relatively thermostable protein. Hum j PR-1 shared a similar 3-dimensional folding pattern with other homologous allergens, which was a unique αβα sandwich structure containing 4 α-helices and 6 antiparallel β-sheets, encompassing 4 conserved CAP domain. ConclusionThe natural PR-1 was firstly purified and characterized as a major allergenic allergen in Humulus japonicus pollen. These findings would contribute to developing diagnostic and therapeutic strategies for Humulus japonicus pollinosis.

Epitope-imprinted polydopamine and reduced graphene oxide-based sensing interface for label-free detection of gliadin

Detection of gluten and its components in foods is paramount in ensuring safety of gluten-sensitive individuals. However, the conventional immunodetection methods including gold standard enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction, and advanced proteomic methods suffer from high cost, complicated instrumentation and protocol, and slow assay turnaround time. Herein, we developed an inexpensive electrochemical sensor for gliadin (GLI) detection using electrochemically reduced graphene oxide for sensitivity enhancement and PQQPFPQQ epitope-imprinted polydopamine as selective recognition interface. The sensing performance was improved by optimizing key process factors including template/monomer ratio, pre-polymerization period, electropolymerization and electrochemical reduction cycles, elution time, and rebinding pH and time. The sensor shows a linear relationship between current response and GLI concentration in the 1–15 ppm range with high affinity to the epitope template (imprinting factor IF = 8.68) and target GLI (IF = 3.14), low limits of detection (0. 70 ppm) and quantification (2.33 ppm). The sensor also exhibits excellent selectivity against potential interferents including ovalbumin (selectivity factor SF = 0.15), folic acid (SF = 0.25), and casein (SF = 0.39) in buffered matrix. The obtained sensor-to-sensor reproducibility, repeatability, and stability ensure its sensitive determination of native GLI in rice flour matrix with coefficient of variance in the range of 2.52–4.27 %, and recovery rates of 99.1–101.9 %. The results fare better than those analyzed by traditional ELISA and similar methods. The developed sensor could be used as a reliable platform for rapid detection of allergic gluten components in processed food products.

Allergen characteristics, quality, major allergen content and galenics for mite allergen-specific immunotherapy preparations

SummaryHouse dust mite extracts for allergen-specific immunotherapy (AIT) require in Germany as other common allergens (pollen from sweet grasses [except maize], birch, alder, hazel; bee and wasp venom) marketing authorisation according to the German Therapy Allergen Ordinance (“Therapieallergene-Verordnung”, [TAV]). Mite allergen extracts that have been approved and also those which are in the approval process are subject to government batch testing. Batch test passing is a prerequisite for marketability. Appropriate quality, efficacy, and safety are prerequisites for the approval of house dust mite extracts. Five HDM allergen extracts from four manufacturers are currently approved in Germany for subcutaneous or sublingual therapy; further extracts are in the approval process. The allergen strength of different products is not comparable; manufacturers use company-specific units to describe the strength. Of the three known major allergens (Group 1 allergens: Der p 1, Der f 1, Group 2 allergens: Der p 2, Der f 2 and Group 23: Der p 23, Der f 23) only Group 1 and Group 2 allergens are usually used to standardize the extracts. Group 23 allergens are localized in the outer membrane of mite faeces, and elution requires special extraction methods. To be efficacious in a single patient an allergen extracts used for AIT must contain all allergen components against which the patient is sensitised. Based on post hoc analyses of large clinical studies, it has been proven for house dust mite tablets that Der p 23 is also contained. In Germany, the Paul Ehrlich Institute is responsible for the approval of therapeutic allergens. For the marketing authorisation information on production procedures, and quality are necessary efficacy and safety (positive benefit risk ratio) of the product must be demonstrated in clinical trials according to the current state of the art.

Molecular sensitization patterns of common food-and respiratory allergens in the Hungarian population

BackgroundRecently developed Immunoglobulin-E (IgE) based molecular allergy diagnostics provide the ability of identifying allergenic components or ingredients at the molecular level (component-resolved-diagnosis, CRD). Compared to the classical IgE-based allergy diagnostics, molecular technology is providing more sensitive and specific IgE-sensitization patterns. Certain sensitization patterns are characteristic of large geographic regions. There are only few data available on the molecular IgE sensitization patterns in East-Central Europe. This study aims to present further data from this region. MethodsData of 3993 stored, anonymized molecular ImmunoCap IgE measurements (CRD), performed in Hungary between January-December 2019 from sera of 1288 subjects (mean age: 27 years ±18 years, male/female ratio 0.56) were analyzed retrospectively, in order to get a local distributional pattern of the sensitizing (IgE >0.35 KU/l) molecular allergens. ResultsThe proportion of CRD positive cases was 24.3%. Amongst them, the most prevalent inhalative allergens were Amb a 1 (18%) Art v 1 (8%) in adults and Der p 2 (3%) and Der p 1 (3%) and Amb a 1 (4%) in subjects below 18 years of age. The same for food allergens were Gal d 2 (21%), Bos d 4 (17%), Bos d 5 (11%) in adults and Gal d 2 (38%), Gal d 1 (28%), Bos d 4 (21%), Bos d 5 (13%) and Bos d 8 (7%) in children. The ratio of mono-sensitivities among CRD-positive cases was 37.5%. ConclusionOur results provide region-specific patterns of sensitization and molecular allergen spreading for Hungary. The relatively higher abundance of polysensitization's among allergic cases underlines the need for early diagnostic -and preventive measures in the future.

Fungal sensitization in patients with severe atopic dermatitis as a distinct phenotype

Background. Many factors influence the development of atopic dermatitis (AD): genetic, environmental (including exposure to allergens), skin barrier damage, and activation of the T2-pathway immune response. Patients with AD, including those with severe disease, are prone to sensitization to various allergens, including fungal ones. Fungal sensitization (FS) promotes autoreactivity to the body's own structures due to shared epitopes with homologous fungal allergens. It can contribute to the development of allergic diseases, including AD, asthma, and rhinitis, as well as to their exacerbation and uncontrolled course. Since FS can be considered a factor aggravating AD, it is relevant to distinguish patients with FS and AD into a separate phenotype.
 Aim. To characterize the phenotype of patients with severe AD and FS using retrospective data analysis from a digital analytics platform in a real-world clinical setting.
 Materials and methods. A retrospective observational single-center study was conducted between 01.06.2017 and 01.07.2022. The study cohort included 88 patients with severe AD who were candidates for therapy or received treatment with either dupilumab or upadacitinib. FS was confirmed in 49 patients from the study group. Part of the cohort without FS (n=39) was used as a comparison group. Inclusion criteria: the age over 18 years old; severe AD (SCORAD40 points at the beginning of therapy); determination of specific immunoglobulin E to a panel of fungal allergens or individual fungi (or by the ImmunoCAP ISAC method to fungal allergen components). A digital analytics platform was used to generate primary data.
 Results. The phenotype of a patient with severe AD and confirmed FS was described. The patient profile is characterized by an extended sensitization spectrum (at least 34 allergen groups) with the most typical combination of pollen, epidermal, and FS. If there is a food allergy, it is of the classic "big eight" nature. Besides exacerbation of the skin disorder, its manifestations include angioedema of life-threatening localization and bronchospasm. Markers of T2 inflammation (high levels of immunoglobulin E and blood eosinophilia) are observed according to the test results, and the T2 endotype of allergy-related abnormalities is typical.
 Conclusion. Apparently, the presence of FS in patients with AD may exacerbate parenteral sensitization mechanisms, expanding its spectrum, including food allergens. Therefore, distinguishing the phenotype of severe AD with FS needs further detailing with a subsequent adaptation of monitoring and treatment methods of severe AD.

Oak Pollen Allergy in Korea.

Oak pollen allergy is common all over the world and an important cause of pollinosis. The molecular properties of some component allergens have been clearly characterized, while some of them are still waiting for characterization. Studies on some oak component allergens are neglected, possibly because of its high cross-reactivity to birch. However, the utilization of culprit allergen molecules is expected to increase the diagnostic sensitivity and efficacy of immunotherapy. Sensitization to oak pollen along with birch often causes pollen food allergy syndrome to fruits and vegetables. Acorn and wood dust from oak can cause allergic disease. We summarize the distribution and taxonomic classification of oak trees of allergenic importance. Molecular characteristics of the identified component allergens, cross-reactivity, and clinical aspects for diagnosis and immunotherapy are also described with an emphasis on Korean situations.

The association between specific IgE antibodies to component allergens and allergic symptoms on dog and cat exposure among Korean pet exhibition participants

BackgroundComponent resolved diagnostics (CRD) in dog and cat allergy is not sufficiently investigated, especially regarding new components such as Can f 4, Can f 6, and Fel d 7. The purpose of this study is to evaluate the potential role of CRD with new components in predicting allergic symptoms on dog and cat exposure. MethodsAmong 552 Korean adults who participated in a pet exhibition and completed questionnaires regarding exposure to dog or cat and allergic symptoms, 522 were venipunctured for measurement of IgE and IgG4 antibody concentration against dog and cat dander extract and underwent skin prick test (SPT). In 238 individuals who were sensitized for both dog and cat dander extract, the dog IgE components (Can f 1–6) and the cat components (Fel d 1/2/4/7) were analyzed. ResultsAn increasing number of sensitizing components was associated with the likelihood of having any allergic symptoms (P < 0.001 for dog and P < 0.01 for cat), and those of asthma (P < 0.01 for dog and P < 0.05 for cat) and rhinoconjunctivitis (P < 0.001 for dog and P < 0.05 for cat). Pairwise correlation of IgE levels was r = 0.56 (P < 0.001) for Can f 6 and Fel d 4, r = 0.74 (P < 0.001) for Can f 1 and Fel d 7 and r = 0.84 (P < 0.001) for Can f 3 and Fel d 2. ConclusionsPolysensitization to dog and cat allergen components is associated with high likelihood of having allergic symptoms during exposure to dogs and cats. Cross-reactivity between dog and cat allergen components is also identified. CRD has a potential in fine-tuning prediction for allergic symptoms on dog and cat exposure.

Allergen Profile of London Plane Tree Pollen: Clinical and Molecular Pattern in Central Spain.

Platanus acerifolia (London plane tree) is a deciduous tree of the Platanaceae family. Sensitization to this plant varies with geography. Madrid, located in central Spain, has one of the highest London plane tree pollen concentration levels on the Iberian Peninsula. We evaluated both the clinical characteristics and the molecular sensitization pattern of patients with allergy to London plane tree pollen in the region of Madrid. Thirty-eight patients allergic to London plane tree pollen were selected according to their clinical symptoms and positive results in skin prick testing and/or specific IgE determination. Serum was collected, and allergen components were evaluated using immunodetection techniques as well as ImmunoCAP. The IgE-binding proteins detected were identified and characterized using mass spectrometry. Analysis of serum samples from allergic patients revealed 9 IgE-binding bands in London plane tree pollen extract. Among these, the 45-kDa protein, which corresponded to Pla a 2, was detected in 76.3% of patients. However, the 18-kDa (Pla a 1) and 9-kDa (Pla a 3) bands were detected in 44.7% and 23.7% of sera, respectively. These results were confirmed using purified proteins. Characterization of the allergen revealed the 27-kDa protein to be glutathione-S-transferase. The molecular profile of patients sensitized to London plane tree pollen differs from that reported in studies from other locations. In the population we studied, the prevalence of Pla a 2 was higher than that of Pla a 1 and Pla a 3. In addition, the minor allergen previously referred to as Pla a 4 was characterized as glutathione-S-transferase.

Comparison of sensitization patterns to dust mite allergens between atopic dermatitis patients and dogs, and non-specific reactivity of canine IgE to the storage mite Tyrophagus putrescentiae.

House dust mite is a common cause of atopic dermatitis (AD) both in humans and dogs. Detection of serum IgE to allergens is commonly used to diagnose allergic diseases. However, false-positive reactions due to cross-reactivity and non-specific reactivity may lead to misdiagnosis. We compared human and canine IgE reactivities to mite component allergens. Canine IgE-reactive components of Dermatophagoides farinae and Tyrophagus putrescentiae were identified by tandem mass spectrometry. Recombinant proteins were produced and IgE reactivities to component allergens were assessed by ELISA and inhibition assays using sera from AD patients and dogs. Canine IgE-reactive proteins (Der f 1, Der f 11, Tyr p 4, Tyr p 8, Tyr p 11, Tyr p 28) were identified by proteome analysis. Most patients were sensitized to Der f 1 (93.3%) and Der f 2 (86.7%). Dogs showed high sensitization to Der f 2 (94.1%) and Der f 18 (84.6%). Both patients and dogs showed low IgE binding frequency to Tyr p 8, 43.3% and 4%, respectively. The ELISA inhibition study indicated that canine IgE reactivity to T. putrescentiae is mostly due to non-specific reaction and cross-reaction with D. farinae. Different IgE sensitization patterns were shown between allergic humans and dogs with AD, especially to Der f 18, for the first time in Korea. Furthermore, non-specific canine IgE reactivity to storage mite indicates the possibility of misdiagnoses. Standardizations focused on the major canine allergen content of extracts should be developed. This will allow precision diagnosis and individuated treatments for each patient and atopic dog.

Sensitization to molecular dog allergens in an adult population: Results from the West Sweden Asthma Study.

As the prevalence of dog allergy rises, component resolved diagnosis might improve the diagnosis, understanding of the clinical outcomes and the effectiveness of immunotherapy. Considering the paucity of data in adults, the current study characterized the patterns of sensitization to dog molecular allergens in an adult population. Data were derived from the West Sweden Asthma Study, a population-based and representative sample of adults from western Sweden. Of the 2006 subjects clinically examined, 313 participants sensitized to whole dog allergen extract were measured for specific immunoglobulin E (sIgE) levels to Can f 1, Can f 2, Can f 3, Can f 4, Can f 5 and Can f 6 using ImmunoCAP™. Polysensitization was defined as sensitization to ≥3 components. Overlapping sensitization was defined as having concomitant sensitization to at least two dog molecular allergen families (lipocalin, albumin or prostatic kallikrein). Of 313, 218 (70%) subjects tested positive to at least one dog allergen component. Sensitization to Can f 1 (43%) was the most common, followed by Can f 5 (33%) among molecular allergens, while sensitization to lipocalins (56%) was the most common among component families. Polysensitization was found in 22% of all participants and was more common in participants with than in those without asthma. Subjects with asthma were less likely to be monosensitized to Can f 5 than those without asthma. Subjects with asthma had higher IgE levels of Can f 3, Can f 4 and Can f 6 than those without asthma. Overlapping sensitizations also differed between those with asthma and allergic rhinitis and those without. Increased knowledge about the sensitization patterns of dog allergen components can aid in defining their role in asthma and rhinitis. In complex clinical cases of dog allergy, a detailed analysis of dog allergen components can provide additional information on the nature of sensitization.

Reconsider the Art of Allergen Immunotherapy

This symposium took place during the European Academy of Allergy and Clinical Immunology (EAACI) Hybrid Congress, held in Prague, Czechia, in July 2022. Stephanie Dramburg, Medical Doctor at Charité Universitätsmedizin, Berlin, Germany, explained that the traditional diagnostic work-up for allergic rhinoconjunctivitis (AR) comprises retrospective clinical history and allergen exposure assessment, extract-based diagnostics, component-resolved diagnostics (CRD), which enables markers of genuine sensitisation to be distinguished from markers of cross-reactivity, and confirmation of clinical relevance. She highlighted that molecular IgE assessment supports the diagnostic work-up and personalised risk assessment in complex cases and that confirmation of clinical relevance of IgE results is key. Furthermore, broadening of a serum IgE response is indicative of ‘molecular spreading’. Dramburg considered that digital technologies have the potential to enhance medical decisions at the point-of-care via targeted patient information, guideline- and evidence-based clinical knowledge, and prospectively collected patient- and sensor-generated data. Marek Jutel, Medical Professor at Wroclaw Medical University, Poland, and the ALL-MED Medical Research Institute, Wroclaw, Poland, described that patients with allergies show different clinical pictures due to differing sensitisation profiles assessed at the molecular level. He noted that patients with allergies react differently to different allergen doses and allergen immunotherapy (AIT) preparations, and minor/intermediate allergens are necessary, in addition to major allergens, for optimal clinical efficacy. Jutel described that allergens that are decisive for AIT efficacy are defined in grass pollen but are not yet determined for other allergen sources such as birch and house dust mite (HDM). Petra Zieglmayer, Medical Professor at Karl Landsteiner University, Krems, Austria, and Head of Vienna Challenge Chamber, Austria, discussed that patients with allergies show complex molecular sensitisation profiles and that extract preparations from different manufacturers vary in terms of allergen composition, with major and intermediate allergens not always detectable. She clarified that optimal efficacy of AIT may only be expected from preparations containing all relevant allergen components in sufficient amounts. Zieglmayer proposed that the target should be to find a match between the patient molecular sensitisation profile and the allergen preparation and that this can be achieved.

Reconsider the Art of Allergen Immunotherapy

This symposium took place during the European Academy of Allergy and Clinical Immunology (EAACI) Hybrid Congress, held in Prague, Czechia, in July 2022. Stephanie Dramburg, Medical Doctor at Charité Universitätsmedizin, Berlin, Germany, explained that the traditional diagnostic work-up for allergic rhinoconjunctivitis (AR) comprises retrospective clinical history and allergen exposure assessment, extract-based diagnostics, component-resolved diagnostics (CRD), which enables markers of genuine sensitisation to be distinguished from markers of cross-reactivity, and confirmation of clinical relevance. She highlighted that molecular IgE assessment supports the diagnostic work-up and personalised risk assessment in complex cases and that confirmation of clinical relevance of IgE results is key. Furthermore, broadening of a serum IgE response is indicative of ‘molecular spreading’. Dramburg considered that digital technologies have the potential to enhance medical decisions at the point-of-care via targeted patient information, guideline- and evidence-based clinical knowledge, and prospectively collected patient- and sensor-generated data. Marek Jutel, Medical Professor at Wroclaw Medical University, Poland, and the ALL-MED Medical Research Institute, Wroclaw, Poland, described that patients with allergies show different clinical pictures due to differing sensitisation profiles assessed at the molecular level. He noted that patients with allergies react differently to different allergen doses and allergen immunotherapy (AIT) preparations, and minor/intermediate allergens are necessary, in addition to major allergens, for optimal clinical efficacy. Jutel described that allergens that are decisive for AIT efficacy are defined in grass pollen but are not yet determined for other allergen sources such as birch and house dust mite (HDM). Petra Zieglmayer, Medical Professor at Karl Landsteiner University, Krems, Austria, and Head of Vienna Challenge Chamber, Austria, discussed that patients with allergies show complex molecular sensitisation profiles and that extract preparations from different manufacturers vary in terms of allergen composition, with major and intermediate allergens not always detectable. She clarified that optimal efficacy of AIT may only be expected from preparations containing all relevant allergen components in sufficient amounts. Zieglmayer proposed that the target should be to find a match between the patient molecular sensitisation profile and the allergen preparation and that this can be achieved.

Topical Application of Metal Allergens Induces Changes to Lipid Composition of Human Skin.

Lipids are an important constituent of skin and are known to be modified in many skin diseases including psoriasis and atopic dermatitis. The direct effects of common metallic contact allergens on the lipid composition of skin has never been investigated, to the best of our knowledge. We describe skin lipid profiles in the stratum corneum and viable epidermis of ex vivo human skin from a female donor upon exposure to three metal allergens (nickel, cobalt and chromium) visualised using time-of-flight secondary ion mass spectrometry (ToF-SIMS), which allows for simultaneous visualisation of both the allergen and skin components such as lipids. Multivariate analysis using partial least squares discriminant analysis (PLS-DA) indicated that the lipid profile of metal-treated skin was different to non-treated skin. Analysis of individual ions led to the discovery that cobalt and chromium induced increases in the content of diacylglycerols (DAG) in stratum corneum. Cobalt also induced increases in cholesterol in both the stratum corneum and viable epidermis, as well as monoacylglycerols (MAG) in the viable epidermis. Chromium caused an increase in DAG in viable epidermis in addition to the stratum corneum. In contrast, nickel decreased MAG and DAG levels in viable epidermis. Our results indicate that skin lipid content is likely to be altered upon topical exposure to metals. This discovery has potential implications for the molecular mechanisms by which contact allergens cause skin sensitization.

Sensitisation to molecular components of mites in atopic dermatitis patients

ABSTRACT Background and aim: To analyse the sensitisation profile to molecular components of mites allergens with the use of ALEX2 Allergy Explorer test in Atopic dermatitis (AD) patients. Method: The complete dermatological and allergological examination including the examination of the sensitisation to molecular components with ALEX2 Allergy Explorer testing was performed. For the statistical analysis, we used Fisher's Exact. Results and conclusion: The sensitisation to mites increases with AD severity. The central role in atopic march may play the molecular component Der f 2 and Der p 23. The results of our study confirm the important role of Der p 5 and Der p 7 in subgroup of patients suffering from bronchial asthma. The specific immunotherapy should focus on the components of NPC2 family and Der p 23 in AD patients. Our results may be helpful in planning hyposensitisation, treatment and immunotherapy.

Profiles of IgE sensitization to dust mite allergen components in patients with allergic rhinitis and asthma

Objective:To study the differences and clinical significance of dust mite allergen components in allergic rhinitis（AR） and allergic rhinitis with asthma syndrome（ARAS） patients. Methods:The clinical data of 42 AR patients were retrospectively analyzed and patients were divided into AR and ARAS group. The serum sIgE concentrations of house dust mites were detected by ImmunoCAP system. The allergen components of Der p（Der p 1, Der p 2, Der p 7, Der p 10, Der p 21, Der p 23） and Der f（Der f 1, Der f 2） were analyzed by protein microarray method. The concentration differences of dust mite allergen and its components in AR and ARAS groups were analyzed. Results:Thirty-one cases of AR and 11 cases of ARAS were included. The positive rate of Der p and Der f was 100.0% and 97.6%, respectively. The highest sensitization rates of Der p allergen components were as following: Der p 1（73.8%）, Der f 1（66.7%）, Der f 2（64.3%） and Der p 2（61.9%）. The sensitization rates of Der f 1（100.0% vs 54.8%, P=0.006）, Der p 2（90.9% vs 51.6%, P=0.021） and Der f 2（100.0% vs 51.6%, P=0.004） in ARAS group were significantly higher than those in AR group. The sIgE concentrations of Der p in AR group were significantly lower than those in ARAS group（[7.65±12.15]kUA/L vs[15.20±18.77]kUA/L, P<0.05）. The sIgE concentrations of Der p 1（[5.39±4.61]kUA/L vs[2.03±2.97]kUA/L, P=0.013）, Der p 2（[8.82± 13.58]kUA/L vs[2.78±5.80]kUA/L, P=0.001）, Der p 23（[1.76± 3.88]kUA/L vs[0.28±0.65]kUA/L, P<0.001） was significantly higher in ARAS group than that of AR group. Correlation analysis showed that Der p 1, Der p 2, Der f 1 and Der f 2 had high positive correlation（P<0.01）. The dust mite components sensitization showed a multiple-sensitized mode. 66.7% of the 42 patients were positive for two or more components while it was 58.1% of the AR group and 90.9% of the ARAS group. The sensitization rate of 3 or more components in ARAS group was significantly higher than that in AR group（54.6% vs 29.1%, P<0.05）. Conclusion:The concentration of dust mites allergens in ARAS group is higher than that in AR group. Der p 1, Der f 1, Der p 2 and Der f 2 are the main allergen components with a higher sensitization rate in ARAS group. The concentrations of Der p 1, Der p 2 and Der p 23 were higher in ARAS group. The ARAS group is prone to multi-sensitzed to allergen components.