Commercial Allergen Extracts Research Articles

Distinct proteomes and allergen profiles appear across the life-cycle stages of Alternaria alternata

Alternaria alternata is associated with allergic respiratory diseases, which can be managed with allergen extract-based diagnostics and immunotherapy. It is not known how spores and hyphae contribute to allergen content. Commercial allergen extracts are manufactured by extracting proteins without separating the different forms of the fungus. We sought to determine differences between spore and hyphae proteomes and how allergens are distributed in Aalternata. Data-independent acquisition mass spectrometry was used to quantitatively compare the proteomes of asexual spores (nongerminating and germinating) with vegetative hyphae. We identified 4515 proteins in nongerminating spores, germinating spores, and hyphae; most known allergens are more abundant in nongerminating spores. On comparing significant protein fold-change differences between nongerminating spores and hyphae, we found that 174 proteins were upregulated in nongerminating spores and 80 proteins in hyphae. Among the spore proteins are ones functionally involved in cell wall synthesis, responding to cellular stress, and maintaining redox balance and homeostasis. On comparing nongerminating and germinating spores, 25 proteins were found to be upregulated in nongerminating spores and 54 in germinating spores. Among the proteins specific to germinating spores were proteases known to be virulence factors. One of the most abundant proteins in the spore proteome is sialidase, which has not been identified as an allergen but may be important in the pathogenicity of this fungus. Major allergen Alt a 1 is present at low levels in spores and hyphae and appears to be largely secreted into growth media. Spores and hyphae express overlapping but distinct proteomes. Most known allergens are found more abundantly in nongerminating spores.

Sensitization to Rice in Allergic Children Pediatric Allergy and Immunology Unit, Children's Hospital, Ain Shams University

Background The prevalence of food allergy among Egyptian children is largely unknown, more so for the different food allergens. Although there are several studies reporting on the common food allergens, yet none was done to explore the allergy to rice, a food allergen that is much less recognized both by pediatricians and allergic patients. Objectives To estimate the frequency of sensitization to rice in pediatric allergic patients in Egypt. Patients and Methods This analytical cross-sectional study was carried out in the period from April 2019 to September 2020. The study included 150 physician-diagnosed allergic patients aged 6 months to 18 years, enrolled consecutively from the Pediatric Allergy and Immunology Unit, Children's Hospital, Ain Shams university. They were 90 (60%) males and 60 (40%) females. SPT was done using a commercial rice allergen extract, Prick-to-prick test with cooked rice. Patients with clinical and /or SPT results suggestive of rice allergy were subjected to oral challenge test under medical supervision in hospital. Results None of the screened allergic children (n = 150) gave a self/parentally reported history of allergy to rice, nor after inquiry by the investigator. SPT using commercial rice extract revealed that only 8 reacted positively (>3mm) and were considered sensitized. PPT was done to all patients using a home prepared cooked rice extract and the test was positive in 5 patients; 2 of these were positive also by SPT. A total of 11 patients (7.3%) were considered sensitized and were subjected to OCT and none had a positive test. There was statistically significant difference in age between rice sensitized and non sensitized, being higher in rice-sensitized patients, with p-value = 0.043. Conclusion Our study is a pilot research which draws the attention to the existence of sensitization to rice in allergic children. PPT is a useful test with a reliable specificity which when positive, would aid in diagnosing rice allergy in settings with limited resources and unavailability of rice protein extracts.

Comparison of commercial allergen extract with various milk products of skin prick tests in diagnosing cow's milk protein allergy

Introduction: In this study, we aimed to solve the incompatibilities between the skin prick test (SPT) result and the clinical picture encountered among patients. Thus, we used various milk products (raw, ultrahigh temperature [UHT], and pasteurized cow's milk) with commercial milk allergen extract in patients we think may have cow's milk protein allergy (CMPA) and made a comparison among the milk types. Methods: This study was conducted retrospectively between February 1, 2019, to June 1, 2019, at a pediatric allergy polyclinic of the university training/research hospital. A pair of 79 individuals, as control and patient groups, ages between 1 month and 21 years who were presented with a suspicion of CMPA and defined with SPT and/or allergen-specific immunoglobulin E (IgE) levels were included in the study. Allergen-specific IgE tests (specific IgE cow's milk, β-lactoglobulin, α-lactalbumin, casein) were evaluated. SPT was performed with fresh/raw, UHT, and pasteurized (daily) cow's milk besides the standard used commercial extract. Results: In the group included in the study and defined as IgE-mediated CMPA, 52.9% of the 79 patients were boys (n = 46), 47.1% were girls (n = 33), and the median age was 6 months (5.0-8.0 months). In the group of specific IgE cow's milk reaction that was sought, frequency and wheal (induration) size of SPT reaction to fresh/raw milk was also found to be statistically significant between positive- and negative-reaction groups, not detected against other milk products. The area under the curve for the diagnostic value of fresh milk was 93.1%. For the cutoff value of 2.50 mm, 88.1% sensitivity and 90.8% specificity were determined. In the group of specific IgE β-lactoglobulin reaction searched, there was a statistically significant difference in the frequency of SPT positivity and wheal size against UHT, fresh, and daily/pasteurized milk between positive and negative reaction groups. When the correlations of the mean induration axis in the tests were examined, it was found that, among raw milk, daily milk, cow's milk allergen extract, and UHT milk, a strong positive correlation was also statistically significant. Conclusion: The positivity that could not be detected with SPT when using allergen extract in a patient with positive specific IgE cow's milk result in the clinic can be detected with SPT made with fresh (raw) milk, which can be an alternative. In addition, specific IgE β-lactoglobulin seems to be the most compatible with the SPT wheal size (induration) of other milk types rather than extract. Comparative studies of SPT allergen extracts and various milk types used in the diagnosis of CMPA are very scarce in the literature, and more studies are needed on this subject.

Prevalence of rat and mouse sensitization in Asian patients with respiratory allergy.

Skin prick test (SPT) is useful in identifying rat and mouse sensitization. To determine the prevalence of rat and mouse sensitization by using local and commercial allergen extracts. Patients with allergic rhinitis or asthma were recruited. SPT of local and commercial rat and mouse allergen extracts were performed. The level of rat and mouse specific IgE (sIgE) was quantified in all patients with positive SPT and randomized patients with negative SPT. Two hundred and thirty patients, 108 male (47%) and median age 14 years (3.2-63.5 years), were enrolled. Rat sensitization by SPT was 11.7% and mouse sensitization was 17.8%. SPT result to local rat and commercial rat allergen extracts were moderately correlated (rs = 0.51, p < 0.001), while SPT result to local mouse and commercial mouse allergen extracts showed low correlation (rs = 0.38, p < 0.001). The concordance of SPT results between local rat and commercial rat allergen extracts was 90.4%. Concordance between the local mouse and commercial mouse allergen extracts was 85.2%. When compared with rat and mouse sIgE, the concordance of local rat, commercial rat and commercial mouse allergen extract were > 80% while that of local mouse was 54.4%. No adverse effect was observed in SPT with any allergen or extract. The prevalence of rat and mouse sensitization was low compared to the study in USA. SPT with local rat and mouse allergen extract was safe and showed good concordance with the SPT result of commercial allergen extracts and rat and mouse sIgE levels.

Sensitisation to Imbrasia belina (mopane worm) and other local allergens in rural Gwanda district of Zimbabwe

BackgroundThe prevalence of allergic diseases is increasing in Zimbabwe and the data relate to local as well as exotic allergen sources. As entomophagy, the practice of eating insects, is a recognised source of local allergens, we sought to measure the prevalence of and risk factors for sensitisation to Imbrasia belina (mopane worm), a popular edible insect. This was investigated alongside other locally relevant allergens in a rural community in Gwanda district, south of Zimbabwe.MethodsA cross sectional study was conducted among 496 adults and children aged 10 years and above in Gwanda district, a mopane worm harvesting area in Zimbabwe. Data on individual characteristics and mopane worm exposure factors were collected using questionnaires. Sensitivity to allergens was assessed by performing skin prick tests at a local clinic using 10 different commercial allergen extracts (Stallergenes, France) and in-house extracts of mopane worm (Imbrasia belina) and mopane leaves (Colophospermum mopane). Data were analysed using Stata version 13 software.ResultsThe prevalence of sensitisation to at least one allergen was 31.17% (n = 144). The prevalence of atopy was higher in adults (33.33%) than in children (23.53%) (p = 0.059). The commonest inhalant allergen sources were mopane worm (14.29%), Tyrophagus putrescentiae (14.29%), mopane leaves (13.42%), Alternaria alternata (6.49%) and Dermatophagoides pteronyssinus (6.49%). Polysensitisation was demonstrated in the study population and of the 108 participants (75%) who were sensitised to two or more allergens, 66 (61%) were women. Sensitisation to mopane worm and mopane leaves often clustered with Tyrophagus putrescentiae amongst adults. Adjusted logistic regression analyses between mopane worm sensitisation and self-reported exposure variables showed that sensitisation was more likely amongst mopane worm harvesters (OR = 1.92, 95%CI = 0.77–4.79), those who cooked or roasted mopane worms during harvesting (OR = 2.69, 95%CI = 0.78–9.31) and harvesting without personal protective equipment (PPE) (OR = 2.12, 95%CI = 0.83–5.44) compared to non-harvesters.ConclusionAtopic sensitization was common in this mopane worm harvesting community in Gwanda district of Zimbabwe. There was frequent co-sensitisation of mopane worm and mopane leaves with Tyrophagus putrescentiae in children and adults. It is important to determine the clinical relevance of our findings, particularly relating to mopane worm sensitisation.

Proteomic evaluation of Alternaria alternata spores, hyphae, and commercial allergen extracts

Alternaria alternata is associated with allergic respiratory diseases, which can be managed with allergen extract-based diagnostics and immunotherapy. It is not known how asexual spores or the vegetative hyphae contribute to patient allergen reactivity and commercial allergen extracts are manufactured by extracting proteins without separating these components. Immunotherapy and diagnosis with existing products, while safe and effective, could be improved with better characterization of these elements.

Homemade Food Allergen Extracts for Use in Skin Prick Tests in the Diagnosis of IgE-Mediated Food Allergy: A Good Alternative in the Absence of Commercially Available Extracts?

Introduction: The skin prick test (SPT) is the first step in the diagnosis of an immunoglobulin E (IgE)-mediated food allergy. The availability of commercial food allergen extracts is very limited, resulting in a need for alternative extraction methods of food allergens. The objective of this study was to compare the SPT results of homemade food allergen extracts with commercially available extracts. Methods: Adult patients with a suspected food allergy were included. Food allergen-specific symptoms were scored using a questionnaire. SPTs were performed with homemade and commercially available extracts (ALK-Abelló, Kopenhagen, Denmark) from almond, apple, hazelnut, peach, peanut, and walnut. Serum-specific IgE was measured with ISAC or ImmunoCAP™. Intra-class correlation coefficients (ICC) between the SPT results of both extract methods were calculated. The proportion of agreement with food allergen-specific symptoms was analyzed. Results: Fifty-four patients (mean age 36; range 19–69 years; female/male: 42/12) were included. The intra-class correlation coefficient (ICC) between the SPT results of both extract methods were strong for hazelnut 0.79 (n = 44) and walnut 0.78 (n = 31), moderate for apple 0.74 (n = 21) and peanut 0.66 (n = 28), and weak for almond 0.36 (n = 27) and peach 0.17 (n = 23). The proportion of agreement between SPT results and food allergen-specific symptoms was comparable for homemade and commercially available extracts, except for peach; 0.77 versus 0.36, respectively. Conclusion: In the diagnostic procedures to identify an IgE-mediated food allergy, homemade extracts from hazelnut and walnut appear to be a good alternative in the absence of commercially available food allergen extracts.

Diversity and complexity of mouse allergens in urine, house dust, and allergen extracts assessed with an immuno-allergomic approach.

Mouse allergy is an important cause of indoor asthma and allergic rhinoconjunctivitis. The major mouse allergen, Mus m 1, is a complex of homologous pheromone-binding lipocalins called major urinary proteins (MUPs). We analyzed the proteome of MUPs in mouse urine, commercial mouse epithelial extracts, and environmental samples using several approaches. These include as follows: two-dimensional electrophoresis and immunoblotting; liquid chromatography-high-resolution mass spectrometry (LC/HRMS); multiple reaction monitoring (MRM) mass spectrometry; and LC/HRMS analysis of glycans at the N-66 residue of MUP3. Albumin is predominant in the extracts, while MUPs are predominant in urine. LC/HRMS of 4 mouse allergen extracts revealed surprising heterogeneity. Of 22 known mouse MUPs, only 6 (MUP3, MUP4, MUP5, MUP13, MUP20, and MUP21) could be identified with MRM using unique peptides. Assessment of MUP content in urine, extracts, and dust samples showed good correlation between MRM and other methods working with different detection principles. All 6 identifiable MUPs were found in electrophoretically separated urine bands, but only MUP3 and MUP20 were above LOQ in unseparated mouse urine, and only MUP3, MUP4, and MUP20 were found in mouse epithelial extracts. Glycan heterogeneity was noted among 4 individual inbred mice: of 13 glycan structures detected, 8 were unique to one mouse, and only 2 glycan modifications were present in all 4 mice. Using mass spectrometry and MRM, mouse allergen extracts and urine samples are shown to be complex and heterogeneous. The efficacy and safety of commercial mouse allergen extracts will be improved with better controls of allergen content.

Proteomic evaluation of Alternaria alternata spores, hyphae, and commercial allergen extracts

Alternaria alternata is associated with allergic respiratory disease which has led to the need for allergen extract-based immunotherapies and diagnostics. Available commercial Alternaria allergen extracts are neither standardized nor well-characterized with regard to allergen content. Immunotherapy and diagnosis with existing products, while safe and effective, could be improved with better characterization and manufacturing consistency. The goal of this study is to apply analytical methods, including quantitative mass spectrometry, for comparative and comprehensive characterization of Alternaria allergen extracts from various source materials.

Prick and intradermal skin tests in patients with severe hymenoptera sting allergy using commercial versus in-house allergen extracts.

Fire ant, honey bee, and wasp allergen extracts are useful in the diagnosis and treatment of severe Hymenoptera allergic patients. To evaluate the result of skin prick test (SPT) and intradermal test (ID) compared between local and commercial insect allergen extracts in patients with severe Hymenoptera sting allergy. SPT and ID using local and commercial insect allergen extracts were performed. Specific IgE (sIgE) to honey bee, wasp, and fire ant; component-resolved diagnosis (CRD); (rApi m1, rApi m2, rApi m3, rApi m5, rApi m10, rVes v5, rPol d5, and rVes v1); and, cross-reactive carbohydrate determinant (CCD) were performed. Twenty-seven patients were included. Twenty-five had anaphylaxis, and 2 had severe systemic skin reaction. Positive skin test (SPT and/or ID) result from local and commercial allergen extracts was 74% vs. 67% for fire ant, 48% vs. 59% for honey bee, and 52% vs. 74% for yellowjacket. Local and commercial allergen extracts showed substantial agreement for fire ant (k = 0.647, p = 0.001) and honey bee (k = 0.632, p = 0.001), and moderate agreement for wasp (k = 0.547, p = 0.001). When compared with sIgE subtracted with CCD and/or CRD, skin test results of local fire ant allergen extract showed higher sensitivity (87% vs. 67%), specificity (42% vs. 33%), and accuracy (67% vs. 52%) than commercial extract. Commercial honey bee and wasp showed higher sensitivity (62% vs. 50%, 85% vs. 65%) and accuracy (63% vs. 52%, 78% vs. 70%), respectively. SPT and ID with local or commercial insect venoms could help in confirming and/or identifying the causative insects.

Measurement of German cockroach allergens and their isoforms in allergen extracts with mass spectrometry.

Allergen extracts are the primary tool for diagnosis and treatment of allergic diseases. In the United States, most allergen extracts are non-standardized. More sophisticated analytical approaches are needed to characterize these products and enable manufacturers and regulators to better determine potency. To expand the multiple reaction monitoring (MRM) assay for an in-depth characterization of German cockroach (GCr; Blattella germanica) allergen extracts. We applied advanced liquid chromatography (LC) and mass spectrometry (MS) techniques including MRM. The expanded LC/MRM-MS method was optimized to measure known GCr allergens and their isoforms/variants in commercial extracts and environmental samples. We performed isoform-specific allergen measurements in multiple extracts from four commercial sources and extracts prepared using environmental samples from urban homes. To investigate causes of heterogeneity, we examined over 30 extraction process variables. Evaluation of the commercial extracts confirmed the variability of production lots and commercial sources. Commonly used defatting and extraction protocols yielded extracts with comparable allergen profiles and content. However, the identity and quality of source materials was a major contributor to variability. In comparing commercial GCr extracts to environmental samples, relative quantities of Bla g 1, Bla g 2, Bla g 3, Bla g 4 and Bla g 11 were similar, while Bla g 5, Bla g 6, Bla g 7 and Bla g 8 were present in the environmental samples but largely absent for the commercial extracts. LC/MRM-MS can be used to measure all known GCr allergens in commercial allergen extracts and environmental samples. Significant differences exist between allergen profiles of commercial extracts and the profiles of environmental samples from dwellings. This analytical platform can serve as a template to achieve better product characterization of similarly complex products.

Comparative analysis of skin prick test with immunoblotting of different manufacturers in the context of determining the sensitization to birch allergen in patients with respiratory allergic diseases

The aim of the study was to evaluate the diagnostic parameters of various methods for determining birch allergen sensitization in patients with respiratory allergic diseases – allergic rhinitis and bronchial asthma. Materials and methods: During the study, 88 patients with allergic rhinitis and/or atopic asthma were examined with three different methods of specific allergic diagnosis (in vivo and in vitro) according to the recommendations of the ethics committee of the Pirogov Vinnitsa National Medical University, all of which were out of acute. Inclusion criteria were a diagnosis of allergic rhinitis (both intermittent and persistent) and/or atopic asthma. The pre-test was conducted according to the classic test method according to the normative documents with commercial allergen extracts. Western blots for the determination of IgE levels were performed using RIDA AllergyScreen test systems (R-Biopharm AG, Darmstadt, Germany) and Euroline (Euroimmun). Results and discussion: Systematic error of measurement results of immunoblot of different manufacturers is equal to 2.27 ku/l, which indicates the presence of a systematic difference. The distribution graph corresponds to the type of absolute systematic error graphs. The standard deviation of the difference was 11.28, which is significantly compared to the values themselves. Thus, the results of the two systems for the determination of specific IgE for birch allergen by the Rida AllergyScreen and Euroline methods are not well matched due to systematic divergence of indicators. There is a good agreement between the results of skin testing with birch allergens and the detection of specific IgE by the Rida AllergyScreen test, and between the results of skin testing with birch allergen and the detection of specific IgE by the Euroline method, there is a moderate agreement between the results.

Comparison of diagnostic parameters of methods for determination of sensitization to allergen D. pteronyssinus in patients with respiratory allergic diseases

The aim of the study was to evaluate the diagnostic parameters of various methods for determining allergy sensitization to the dust mite allergen Dermatophagoides pteronyssinus in patients with respiratory allergic diseases – allergic rhinitis and bronchial asthma. Materials and methods: During the study, 88 patients with allergic rhinitis and / or atopic asthma were examined with three different methods of specific allergic diagnosis (in vivo and in vitro) according to the recommendations of the ethics committee of the Pirogov Vinnitsa National Medical University, all of which were out of acute period. Inclusion criteria were a diagnosis of allergic rhinitis (both intermittent and persistent) and / or atopic asthma. The prick-test was conducted according to the classic test method by normative documents with commercial allergen extracts. Western blots for the determination of IgE levels were performed using RIDA AllergyScreen test systems (R-Biopharm AG, Darmstadt, Germany) and Euroline (Euroimmun). Results and discussion: The results of two systems for the determination of specific IgE to D. Pteronyssinus by the Rida AllergyScreen and Euroline methods have a systematic difference of values (-1.27 kU / l). There is a good agreement between the results of skin testing with D. Pteronyssinus allergens and the detection of specific IgE by the Rida AllergyScreen method, and there is a satisfactory agreement between the results of skin testing with D. Pteronyssinus allergens and detection of specific IgE by the Euroline method. According to the results of the curve of detection of specific IgE by AllergyScreen method for determination of sensitization to allergen D. Pteronyssinus has excellent diagnostic significance (AUC=0,953), detection of specific IgE by Euroline method for determination of sensitization to allergen D. Pteronyssinus – good AUC=0,7.

Sensitization to rye allergen in patients with allergic rhinitis and atopic bronchial asthma: comparison of the diagnostic parameters

The aim of the study was to evaluate the diagnostic parameters of various methods for determining sensitization to rye allergen in patients with respiratory allergic diseases — allergic rhinitis and bronchial asthma.Material and methods. In the study, 88 patients with allergic rhinitis and/or atopic asthma were examined with three different methods of specific allergic diagnosis (in vivo and in vitro). Inclusion criteria were a diagnosis of allergic rhinitis (both intermittent and persistent) and/or atopic asthma. The pre-test was conducted according to the classic test method according to the normative documents with commercial allergen extracts. Western blots for the determination of IgE levels were performed using RIDA AllergyScreen test systems (R-Biopharm AG, Darmstadt, Germany) and Euroline (Euroimmun).Results. The results of the two systems for the determination of specific IgE for rye allergen by the Rida AllergyScreen and Euroline methods are not in good agreement due to the significant systematic divergence of indicators. The systematic error of the measurement results is 2.6 ku/l, which indicates the presence of a systematic difference. The distribution graph corresponds to the type of absolute systematic error graphs. The standard deviation of the differences was 8.1, which is significantly compared to the values themselves; there is no dependence of the measurement difference on the amount of specific IgE in the blood.Conclusions. The results of the two systems for the determination of specific IgE for rye allergen by the Rida AllergyScreen and Euroline methods are not in good agreement due to the significant systematic divergence of indicators. Between skin test with rye allergens and detection of specific IgE by the Rida AllergyScreen method there is a moderate agreement between study results, between skin test with rye allergen and detection of specific IgE by the Euroline method there is moderate agreement between test results.

Allergen content in mouse allergenic samples and products is more complex than a single Mus m 1 value

Mus m 1 is a group of >21 polymorphic major urinary proteins (MUPs) and the specificity of available Mus m 1 immunoassays to any of these MUPs is unknown. In order to improve the diagnostic and therapeutic potency of commercial mouse allergen extracts, we need improved methods to characterize mouse allergens and the immune responses to them. We applied the following analytical methods to commercial extracts, urine from laboratory and wild mice, and environmental samples from homes of Baltimore asthma patients: isoelectric focusing, two-dimensional electrophoresis (2D-PAGE), liquid chromatography, immunostaining, mass spectrometry, and multiple reactions monitoring (MRM). By 2D-PAGE with immunostaining, MUPs are the major IgE-binding proteins (90-95%) in urine, but albumin is more abundant in commercial extracts. MUPs have complex migration patterns on 2D-PAGE, attributable to sequence polymorphisms and posttranslational modifications. For example, we identified at least 14 distinct MUP3-variants with variable glycosylation states. Using MRM, we determined that MUPs with >97% sequence similarity constitute 85-90% of the total MUPs in both commercial extracts and urine. Environmental dust samples, from bed, bedroom, and kitchen, contained MUP profiles consistent with those found in commercial extracts. Commercial extracts are highly variable, with up to 30-fold differences in specific MUP content. Using a full array of proteomic and mass spectrometric tools, we can assess the allergen content of commercial mouse allergen extracts and compare it to allergens in mouse urine and environmental dust samples. These tools will help guide further research, characterization, and standardization of mouse allergenic products.

Principles of Allergen Immunotherapy and Its Clinical Application in China: Contrasts and Comparisons with the USA.

Allergen immunotherapy (AIT) for allergic rhinitis (AR), asthma, and other allergic diseases has developed quickly. House dust mite (HDM), Artemisia (wormwood), Humulus japonicus (Japanese hop), Alternaria alternata, and Cladosporium herbarum are the five most common inhalant allergens in China. AIT has been performed in China for over 60years. With the support of the Chinese Medical Association (CMA) and the Chinese Medical Doctors Association (CMDA), the Chinese College of Allergy and Asthma (CCAA) was established in 2016 as a specialized branch of CDMA and is the main certification authority for AIT. Chinese allergists and scientists have made tremendous progress in the development of AIT. There have been many publications by Chinese allergists and scientists worldwide encompassing original research studies, systematic reviews, case studies, and clinical trials. Currently, conventional subcutaneous immunotherapy (SCIT) is the preferred AIT in China, but sublingual immunotherapy (SLIT) is beginning to gain recognition. An increasing number of clinical trials have been conducted to investigate the clinical efficacy and side effects of SLIT and SCIT. In China, HDM is the only commercial standardized allergen extracts in clinical use, whereas the others are crude allergen extracts. Besides standardized allergen extracts, other forms of hypoallergenic extracts are still being investigated and developed in China. Immunotherapy in China is similar to that in the USA in which allergen extracts can be mixed for SCIT. However, allergen extracts cannot be mixed for SCIT in Europe.

Variability of allergens in commercial fish extracts for skin prick testing.

Commercial allergen extracts for allergy skin prick testing (SPT) are widely used for diagnosing fish allergy. However, there is currently no regulatory requirement for standardization of protein and allergen content, potentially impacting the diagnostic reliability of SPTs. We therefore sought to analyse commercial fish extracts for the presence and concentration of fish proteins and invitro IgE reactivity using serum from fish-allergic patients. Twenty-six commercial fish extracts from five different manufacturers were examined. The protein concentrations were determined, protein compositions analysed by mass spectrometry, followed by SDS-PAGE and subsequent immunoblotting with antibodies detecting4 fish allergens (parvalbumin, tropomyosin, aldolase and collagen). IgE-reactive proteins were identified using serum from 16 children with confirmed IgE-mediated fish allergy, with focus on cod, tuna and salmon extracts. The total protein, allergen concentration and IgE reactivity of the commercial extracts varied over 10-fold between different manufacturers and fish species. The major fish allergen parvalbumin was not detected by immunoblotting in 6/26 extracts. In 7/12 extracts, five known fish allergens were detected by mass spectrometry. For cod and tuna, almost 70% of patients demonstrated the strongest IgE reactivity to collagen, tropomyosin, aldolase A or β-enolase but not parvalbumin. Commercial fish extracts often contain insufficient amounts of important allergens including parvalbumin and collagen, resulting in low IgE reactivity. A comprehensive proteomic approach for the evaluation of SPT extracts for their utility in allergy diagnostics is presented. There is an urgent need for standardized allergen extracts, which will improve the diagnosis and management of fish allergy.

Allergy in adolescent population (14-18 years) living in Campania region (Southern Italy). A multicenter study.

Adolescents (Ad) constitute a difficult to manage population among individuals suffering from asthma. The aim of our study was to assess the prevalence, clinical characteristics and age of onset of allergic sensitization and clinical symptoms in a sample of atopic Ad living in the Campania region (Southern Italy). Sixteen Allergy units or Centers belonging to the Italian Association of Hospital and Territorial Allergologists (AAIITO, Campania region) participated in this cross-sectional study. A case report form (CRF) was specifically designed for this study and commercial allergen extracts used for screening SPTs were provided by ALK-Abelló Group (Milan, Italy). A total of 443 patients were examined (females, f 220, 49.6 %; males, m 223, 50.3%). Dust mites represent the most common sensitizing agents in allergic Ad living in Campania region (Dermatoph. pteronyssinus 67.4% and Dermatoph. farinae 66.5%), followed by Parietaria (58.9%), grasses (45.8%), Artemisia vulgaris (16.7%), Olea Europaea (32.2%), dog dander (17.1%), cat dander (20.0%), Alternaria alternata (8.1%), Cupressus sempervirens (4.9%), Betula pendula (4.7%), other allergens (19.4%). An interesting comparison has been made between clinical data of our Ad with data of elderly patients (E). The role of allergic sensitization is significantly higher in Ad compared to E. Dermatophagoides pteronyssinus is the first sensitizing allergen in Ad and the last in E. Parietaria constitutes the first sensitizing pollen both in Ad and E, the percentage of sensitization is higher in Ad. Another important difference is the higher prevalence of As, as only symptom, in E compared to Ad (19.7% versus 7.6%). In conclusion, our findings confirm the high prevalence and clinical significance of airway allergic sensitization in the adolescents living in Campania region.

What Do We Really Know About Intralymphatic Immunotherapy?

Allergen-specific immunotherapy (AIT) is the only causal treatment method of IgE-mediated allergies that may lead to long-term symptom amelioration even after the end of treatment, positively interfere with the course of disease, and improve the immunological situation of the patient. While AIT in general requires treatment periods over 3 up to 5 years, intralymphatic immunotherapy (ILIT) needs only three ultrasound-guided injections of low allergen doses into inguinal lymph nodes with 4-week time interval making the entire treatment possible within 2 months. The number of published ILIT trials is continuously increasing and it has been mainly used for the indication of allergic rhinitis using commercially available grass pollen and birch extracts and Dermatophagoides farinae, Dermatophagoides pteronyssinus, dog or cat allergens, and moreover a recombinant MAT-Fel d 1 vaccine and autologous semen from a patient with post-orgasmic illness syndrome. ILIT is a very promising AIT technique that could widely improve patient treatment However, there is not enough convincing evidence for a routine use of ILIT and no authorized commercial allergen extracts exist for this approach, so far. Dose-escalation studies and prospective DBPC efficacy trials need to be performed in diverse allergens like insect venom. Moreover, pediatric populations have not been present in former ILIT studies.

Underestimation of house dust mite–specific IgE with extract-based ImmunoCAPs compared with molecular ImmunoCAPs

Underestimation of house dust mite–specific IgE with extract-based ImmunoCAPs compared with molecular ImmunoCAPs