Determination of Allergen Levels, Isoforms, and Their Hydroxyproline Modifications Among Peanut Genotypes by Mass Spectrometry

The recently published reference genome of peanuts enables a detailed molecular description of the allergenic proteins of the seed. We used LC-MS/MS to investigate peanuts of different genotypes to assess variability and to better describe naturally occurring allergens and isoforms. Using relative quantification by mass spectrometry, minor variation of some allergenic proteins was observed, but total levels of Ara h 1, 2, 3, and 6 were relatively consistent among 20 genotypes. Previously published RPHPLC methodology was used for comparison. The abundance of three Ara h 3 isoforms were variable among the genotypes and contributed to a large proportion of total Ara h 3 where present. Previously unpublished hydroxyproline sites were identified in Ara h 1 and 3. Hydroxylation did not vary significantly where sites were present. Peanut allergen composition was largely stable, with only some isoforms displaying differences between genotypes. The resulting differences in allergenicity are of unknown clinical significance but are likely to be minor. The data presented herein allow for the design of targeted MS methodology to allow the quantitation and therefore control of peanut allergens of clinical relevance and observed variability

The central role of IL-33/IL-1RL1 pathway in asthma:From pathogenesis to intervention

Interleukin-33 (IL-33), a member of the IL-1 family, and its cognate receptor, Interleukin-1 receptor like-1 (IL-1RL1 or ST2), are susceptibility genes for childhood asthma. In response to cellular damage, IL-33 is released from barrier tissues as an & lsquo;alarmin & rsquo; to activate the innate immune response. IL-33 drives type 2 responses by inducing signalling through its receptor IL-1RL1 in several immune and structural cells, thereby leading to type 2 cytokine and chemokine production. IL-1RL1 gene transcript encodes different isoforms generated through alternative splicing. Its soluble isoform, IL-1RL1-a or sST2, acts as a decoy receptor by sequestering IL-33, thereby inhibiting IL1RL1-b/IL-33 signalling. IL-33 and its receptor IL-1RL1 are therefore considered as putative biomarkers or targets for pharmacological intervention in asthma. This review will provide an overview of the genetics and biology of the IL-33/IL-1RL1 pathway in the context of asthma pathogenesis. It will discuss the potential and complexities of targeting the cytokine or its receptor, how genetics or biomarkers may inform precision medicine for asthma targeting this pathway, and the possible positioning of therapeutics targeting IL-33 or its receptor in the expanding landscape of novel biologicals applied in asthma management. (c) 2021 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license (http:// creativecommons.org/licenses/by/4.0/)

Substructure in the stellar halo near the Sun:I. Data-driven clustering in integrals-of-motion space

Aims: Develop a data-driven and statistically based method for finding such clumps in Integrals of Motion space for nearby halo stars and evaluating their significance robustly. Methods: We use data from Gaia EDR3 extended with radial velocities from ground-based spectroscopic surveys to construct a sample of halo stars within 2.5 kpc from the Sun. We apply a hierarchical clustering method that uses the single linkage algorithm in a 3D space defined by the commonly used integrals of motion energy $E$, together with two components of the angular momentum, $L_z$ and $L_\perp$. To evaluate the statistical significance of the clusters found, we compare the density within an ellipsoidal region centered on the cluster to that of random sets with similar global dynamical properties. We pick out the signal at the location of their maximum statistical significance in the hierarchical tree. We estimate the proximity of a star to the cluster center using the Mahalanobis distance. We also apply the HDBSCAN clustering algorithm in velocity space. Results: Our procedure identifies 67 highly significant clusters ($> 3\sigma$), containing 12\% of the sources in our halo set, and in total 232 subgroups or individual streams in velocity space. In total, 13.8\% of the stars in our data set can be confidently associated to a significant cluster based on their Mahalanobis distance. Inspection of our data set reveals a complex web of relationships between the significant clusters, suggesting that they can be tentatively grouped into at least 6 main structures, many of which can be associated to previously identified halo substructures, and a number of independent substructures. This preliminary conclusion is further explored in an accompanying paper by Ruiz-Lara et al., where we also characterize the substructures in terms of their stellar populations. Conclusions: We find... (abridged version)Comment: 16 pages, 14 figures, 2 tables. Accepted for publication in A&A. This is the first in a series of papers, the second (Ruiz-Lara et al.) can be found in https://ui.adsabs.harvard.edu/abs/2022arXiv220102405R/abstract Code of the clustering algorithm can be found in https://github.com/SofieLovdal/IOM_clusterin

IL-1RL1a serum levels and IL1RL1 SNPs in the prediction of food allergy

Food allergy is a common disorder in the Western world, with increasing prevalence and substantial healthcare costs(1). Food allergy is often accompanied by the presence of specific IgE against harmless proteins in food, but not all sensitized children show clinical reactions upon exposure. Therefore, double-blind placebo-controlled food challenges (DBPCFC) remain the gold standard to diagnose food allergy, yet this test is demanding. Biomarkers that can predict clinical response to food are urgently needed

Sandwich Enzyme-Linked Immunosorbent Assay for Detecting Sesame Seed in Foods

Small amounts of sesame can trigger allergic reactions in sesame-allergic patients. Because sesame is a widely used food ingredient, analytical methods are needed to support quality control and food safety programs in the food industry. In this study, polyclonal antibodies against sesame seed proteins were raised, and an enzyme-linked immunosorbent assay (ELISA) was developed for the detection and quantification of sesame seed residue in food. A comparison was made between this ELISA and other assays, particularly focusing on recovery of sesame seed residue from different food matrices. The developed ELISA is sensitive with a lower limit of quantification of 0.5 ppm and shows essentially no cross-reactivity with other foods or food ingredients (92 tested). The ELISA has a good recovery for analyzing sesame-based tahini in peanut butter, outperforming one other test. In a baked bread matrix, the ELISA has a low recovery, while two other assays perform better. We conclude that a sensitive and specific ELISA can be constructed based on polyclonal antibodies, which is suitable for detection of small amounts of sesame seed relevant for highly allergic patients. Furthermore, we conclude that different food products may require different assays to ensure adequate quantification of sesame

Sandwich Enzyme-Linked Immunosorbent Assay for Detecting Sesame Seed in Foods

Small amounts of sesame can trigger allergic reactions in sesame-allergic patients. Because sesame is a widely used food ingredient, analytical methods are needed to support quality control and food safety programs in the food industry. In this study, polyclonal antibodies against sesame seed proteins were raised, and an enzyme-linked immunosorbent assay (ELISA) was developed for the detection and quantification of sesame seed residue in food. A comparison was made between this ELISA and other assays, particularly focusing on recovery of sesame seed residue from different food matrices. The developed ELISA is sensitive with a lower limit of quantification of 0.5 ppm and shows essentially no cross-reactivity with other foods or food ingredients (92 tested). The ELISA has a good recovery for analyzing sesame-based tahini in peanut butter, outperforming one other test. In a baked bread matrix, the ELISA has a low recovery, while two other assays perform better. We conclude that a sensitive and specific ELISA can be constructed based on polyclonal antibodies, which is suitable for detection of small amounts of sesame seed relevant for highly allergic patients. Furthermore, we conclude that different food products may require different assays to ensure adequate quantification of sesame

Ara h 6 Complements Ara h 2 as an Important Marker for IgE Reactivity to Peanut

The similarities of two major peanut allergens, Ara h 2 and Ara h 6, in molecular size, amino acid sequence, and structure have made it difficult to obtain natural Ara h 6 free of Ara h 2. The objectives of this study were to purify natural Ara h 6 that is essentially free of Ara h 2 and to compare its IgE reactivity and potency in histamine release assays to Ara h 2. SDS-PAGE of the highly purified allergen (\u3c0.01% Ara h 2) revealed a single 14.5kD band and the identity of Ara h 6 was confirmed by LC-MS/MS. Ara h 6 showed a higher seroprevalence in chimeric-IgE ELISA (n=54), but a weaker biological activity in basophil histamine release assays than Ara h 2. Purified Ara h 6 will be useful for diagnostic IgE antibody assays, as well as molecular and cellular studies to investigate the immunological mechanisms of peanut allergy