Human Allergic Diseases Research Articles

A human monoclonal antibody based immunoenzymetric assay to measure Fel d 1 concentrations in cat hair and pelt allergenic extracts.

In the United States, 19 allergen extracts of different specificities are standardized, which means that their potencies are determined in comparison to a US reference standard. For cat allergen extracts, potency is determined by measuring Fel d 1 content expressed in in Fel d 1 units, and with a unitage that correlates with skin test reactions (bioequivalent allergy units or BAU). Currently, Fel d 1 content is measured with a radial immunodiffusion (RID) assay that uses polyclonal sheep antisera to detect the allergenic protein by producing a white precipitin line in agar gel. However, the RID is considered cumbersome, and the polyclonal sera may qualitatively vary among animals and may recognize epitopes irrelevant to human allergic disease. In this report, we describe a quantitative two-site immunoenzymetric assay (IEMA) for Fel d 1 that uses immobilized capture and soluble biotin-labeled detection Fel d 1-specific human IgE monoclonal antibodies (mAb) that have been class-switched to IgG4. Together, they sandwich Fel d 1 molecules from extracts. Using purified natural Fel d 1 as a calibrator, the historically reported ∼4 micrograms Fel d 1/Fel d 1 unit assignment was directly measured in this mAb-based IEMA at 3.12 ± 0.24 micrograms of Fel d 1 per Fel d 1 unit. This IEMA appears to be equivalent to RID in the measurement of biological potencies of commercial cat hair and cat pelt extracts marketed in the United States.

ITRAQ-based proteomics revealed the acaricidal mechanism of methyl isoeugenol against Dermatophagoides farinae

ABSTRACT Dermatophagoides farinae is a major source of allergens that are the cause of allergic diseases in humans. Methyl isoeugenol (MIE) is a naturally occurring compound in plants and its strong acaricidal toxicity has been established; however, the mechanism behind its acaricidal properties remains unclear. In this study, to investigate the action mechanism of MIE on D. farinae the isobaric tags for relative and absolute quantification (iTRAQ) proteomics method is used. Protein samples were extracted from D. farinae before and after 24 h of MIE treatment for iTRAQ technique analysis. Bioinformatics analysis revealed that most differentially expressed proteins were located in the cytoplasm, impacting cytoplasmic function. Additionally, Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses showed significant enrichment of the differential proteins in the cellular autophagy signalling pathway. The autophagy-related proteins Autophagy-Related Protein 3 (ATG3) and Cathepsin L (CTSL) were up-regulated, disrupting the autophagic process. We found that the main action mechanism of MIE against D. farinae might be cellular autophagy. The transcriptional validation of six differential proteins demonstrated consistent expression trends with the proteomics data. This study reveals the acaricidal mechanism of MIE for the first time and lays the foundation for further research on D. farinae control.

TCF1-LEF1 co-expression identifies a multipotent progenitor cell (TH2-MPP) across human allergic diseases.

Repetitive exposure to antigen in chronic infection and cancer drives T cell exhaustion, limiting adaptive immunity. In contrast, aberrant, sustained T cell responses can persist over decades in human allergic disease. To understand these divergent outcomes, we employed bioinformatic, immunophenotyping and functional approaches with human diseased tissues, identifying an abundant population of type 2 helper T (TH2) cells with co-expression of TCF7 and LEF1, and features of chronic activation. These cells, which we termed TH2-multipotent progenitors (TH2-MPP) could self-renew and differentiate into cytokine-producing effector cells, regulatory T (Treg) cells and follicular helper T (TFH) cells. Single-cell T-cell-receptor lineage tracing confirmed lineage relationships between TH2-MPP, TH2 effectors, Treg cells and TFH cells. TH2-MPP persisted despite in vivo IL-4 receptor blockade, while thymic stromal lymphopoietin (TSLP) drove selective expansion of progenitor cells and rendered them insensitive to glucocorticoid-induced apoptosis in vitro. Together, our data identify TH2-MPP as an aberrant T cell population with the potential to sustain type 2 inflammation and support the paradigm that chronic T cell responses can be coordinated over time by progenitor cells.

Fungal Sensitization and Human Allergic Disease.

Fungal sensitizations have been associated with hypersensitivity reactions with variable levels of evidence available to link types of fungi with human disease. We conducted systematic reviews of the literature to identify the strength of evidence linking lesser-studied fungi for which there are commercially available extracts to identify populations in which they were useful in clinical practice. Excluding five fungi for which hundreds of articles were identified, there are 54 articles on the remaining fungi with clinical data. For 12 of the fungi, the prevalence of fungal sensitization varies in different hypersensitivity disorders due to factors related to geographic areas, age, and other underlying medical conditions. There were no studies linking seven genera to human disease. Most of the commercially available fungal extracts are uncommonly associated with hypersensitivity reactions in humans. Specific extracts may be useful in particular disease states such as allergic fungal sinusitis or allergic bronchopulmonary mycosis, or when routine testing fails to identify a cause of uncontrolled disease, such as in asthma.

Lymphatic-Dependent Modulation of the Sensitization and Elicitation Phases of Contact Hypersensitivity

Allergic contact dermatitis is a common inflammatory skin disease comprising 2 phases. During sensitization, immune cells are activated by exposure to various allergens, whereas repeated antigen exposure induces local inflammation during elicitation. In this study, we utilized mouse models lacking lymphatics in different skin regions to characterize the role of lymphatics separately in the 2 phases, using contact hypersensitivity as a model of human allergic inflammatory skin diseases. Lymphatic-deficient mice exhibited no major difference to single antigen exposure compared to controls. However, mice lacking lymphatics in both phases displayed reduced inflammation after repeated antigen exposure. Similarly, diminished immune response was observed in mice lacking lymphatics only in sensitization, whereas the absence of lymphatics only in the elicitation phase resulted in a more pronounced inflammatory immune response. This exaggerated inflammation is driven by neutrophils impacting regulatory T cell number. Collectively, our results demonstrate that skin lymphatics play an important but distinct role in the 2 phases of contact hypersensitivity. During sensitization, lymphatics contribute to the development of the antigen-specific immunization, whereas in elicitation, they moderate the inflammatory response and leukocyte infiltration in a neutrophil-dependent manner. These findings underscore the need for novel therapeutic strategies targeting the lymphatics in the context of allergic skin diseases.

Genetic diversity and differentiation analysis reveals geographical structure characteristics of Dermatophagoides farinae (Acari: Pyroglyphidae).

Dermatophagoides farinae (Acari: Pyroglyphidae) has been reported as one of the major sources of indoor allergens that trigger allergic disease in humans. In this study, the genetic diversity and differentiation of nine geographic populations of D. farinae were investigated by analyzing mitochondrial and nuclear genes (COI, Cytb, COI+Cytb, and ITS). The results showed high genetic diversity across the D. farinae populations. The BX (Benxi) population showed the lowest genetic diversity, possibly due to climatic causes. Significant genetic differentiation was observed among D. farinae populations based on mitochondrial genes. The analysis of molecular variance (AMOVA) results elucidated that the contribution to the rate of variation was primarily from among populations. Phylogenetic analysis and haplotype network based on mitochondrial genes both indicated significant geographic structure among D. farinae populations. The nine geographic populations of D. farinae were divided into two groups with the Qinling Mountains-Huai River Line serving as the boundary for spatial analysis of molecular variance analysis (SAMOVA). However, the Mantel test analysis showed no association between genetic differentiation and geographic distance because of the high level of gene flow among some populations through the transportation of stored food. Overall, these results indicate both significant genetic differentiation among D. farinae populations, but also significant gene exchange between them. Results from the analysis of the nuclear gene ITS differed from the mitochondrial genes due to differences in molecular markers between mitochondrial genes and nuclear genes. These observations improve our understanding of the genetic diversity and structure of D. farinae populations.

Could mosquito bites facilitate IgE sensitization to food and inhalant allergens?

We present an hypothesis that arthropod bites and stings can induce sensitization to concomitantly introduced food and inhalant allergens in human, and that contents of the sting and bite such as venom and saliva proteins act as adjuvants further facilitating sensitization. In food allergen sensitization, evidence presently supports the hypothesis that exposure through a disrupted skin-barrier is allergenic, and exposure to food proteins through ingestion is tolerogenic. Stinging and biting arthropods that breach skin barrier are known to provide IgE sensitization to their own venom and salivary allergens e. g. Hymenoptera and mosquito. Murine experiments have shown that content of venom from honeybee (phospholipase 2 (PLA2) and hyaluronidase) facilitate IgE sensitization to a concomitantly introduced food allergen in the skin. Conserved biomolecules in saliva and venom present in several arthropod species (e.g. PLAs and hyaluronidases) are suggested to play a role as adjuvants in sensitization to concomitantly presented allergens during arthropod stings and bites. Tick bites have been shown to provide IgE sensitization to tick salivary allergens and IgE sensitization to non-primate mammalian red blood cell allergen Galactose-alpha-1,3-galactose, causing allergy to non-primate red meat, termed Alpha-gal syndrome (AGS).Mosquitos may take up allergens such as pathogenesis related proteins (e.g. lipid transfer proteins (LTPs), and PR-10) when feeding on plant nectar and sap and may take up animal derived allergens such as immunoglobulins, serum albumins and lipocalins from feeding on blood. It is proposed that plant and animal allergens, important in human allergic disease are carried on mouthparts or in gut of mosquitos and transferred into the skin of a human host during a mosquito bite. We hypothesize that sensitization to common food and inhalant allergens can occur facilitated by a mosquito bite, possibly facilitated by adjuvant factors such as PLA2 and hyaluronidase in mosquito saliva. The hypothesis may provide insight into basic mechanisms in IgE sensitization.

The re-emerging role of linoleic acid in paediatric asthma.

Asthma is the most common chronic disease within the paediatric population. Although it is multifactorial, its onset may be linked to early-life exposures with subsequent impact on immune system development. Microbial and dietary metabolic products have been implicated in the development and exacerbation of paediatric asthma. Linoleic acid is the most common omega-6 polyunsaturated fatty acid in the Western diet. In this review, we summarise the literature regarding the involvement of linoleic acid in the development of and its impact on existing paediatric asthma. First, we summarise the existing knowledge surrounding the relationship between human microbial metabolism and allergic diseases in children. Next, we examine cellular or animal model-based mechanistic studies that investigated the impact of dietary- and microbial-derived linoleic acid metabolites on asthma. Finally, we review the literature investigating the impact of linoleic acid metabolites on the development and exacerbation of childhood asthma. While there is conflicting evidence, there is growing support for a role of linoleic acid in the onset and pathophysiology of asthma. We recommend that additional cellular, animal, and longitudinal studies are performed that target linoleic acid and its metabolites.

Evolution Toward Chip-Based Arrays in the Laboratory Diagnosis of Human Allergic Disease.

Multiplex-based specific IgE antibody assays have emerged into the clinical immunology laboratory through the combined use of pure, recombinant allergenic molecules and new methods to simultaneously and accurately analyze specific IgE antibodies to hundreds of allergens. This review traces the historical development and examines outstanding questions related to the strengths and limitations of these new molecular allergen multipex technologies for the assessment of human allergic sensitization. Multiplexed technologies are poised to provide the most cost-effective and comprehensive evaluation of patients with suspected allergy as compared with the commonly used singleplex autoanalyzers. How analytically sensitive and quantitative are the multiplex technologies, down to 0.1 kUA/L? Because each allergen is viewed as a unique assay, how will analytical and clinical performance be documented at the manufacturing and clinical laboratory levels to guarantee reproducibility and obtain government regulatory clearance? Will interference by naturally occurring allergen-specific IgG compromise analytical performance? Successful resolution of these and other questions covered in this review will position multiplex technologies to become the single most-effective means of screening patients for allergic sensitization, assessing IgE antibody cross-reactivity, and planning therapy directed at the patient with allergy.

Quantitative control of Ets1 dosage by a multi-enhancer hub promotes Th1 cell differentiation and protects from allergic inflammation

Multi-enhancer hubs are spatial clusters of enhancers present across numerous developmental programs. Here, we studied the functional relevance of these three-dimensional structures in Tcell biology. Mathematical modeling identified a highly connected multi-enhancer hub at the Ets1 locus, comprising a noncoding regulatory element that was a hotspot for sequence variation associated with allergic disease in humans. Deletion of this regulatory element in mice revealed that the multi-enhancer connectivity was dispensable for Tcell development but required for CD4+ T helper 1 (Th1) differentiation. These mice were protected from Th1-mediated colitis but exhibited overt allergic responses. Mechanistically, the multi-enhancer hub controlled the dosage of Ets1 that was required for CTCF recruitment and assembly of Th1-specific genome topology. Our findings establish a paradigm wherein multi-enhancer hubs control cellular competence to respond to an inductive cue through quantitative control of gene dosage and provide insight into how sequence variation within noncoding elements at the Ets1 locus predisposes individuals to allergic responses.

18 Can the Indoor Environment Influence the Toxicity of Indoor and Outdoor Air Pollution Particulates?

Abstract When considering air pollution particles (AP), they are often assessed in isolation, when this is not how the population is exposed to them. Within the indoor environment, there are bioaerosols present, which contain various allergens including house dust mite (HDM) and lipopolysaccharides (LPS). This RESPIRE study aims to apply a well-characterised in vitro model of the alveolar barrier (type I and II epithelial cells (NCI-H441 and hAELVi at relevant in vivo ratios) and macrophage-like cells) to determine the effects of these bioaerosols on the toxicological and immunological responses to AP. To achieve this, we will investigate cytotoxicity, inflammatory responses, morphology, oxidative stress, membrane integrity and genetic toxicology of these models with specific focus on responses from human allergic responses and airway diseases (i.e., bronchiolitis-interstitial lung disease and sarcoidosis) to determine the additional health considerations the addition of bioaerosols to AP has on our models. We will utilise a physiologically relevant aerosolization exposure method) and a standardised indoor (NIST2583) particulate at an environmentally relevant concentration to mimic air pollution along with HDM and LPS to determine if the addition of these environmentally appropriate bioaerosols influence the responses to IAP. This project is funded by the recently initiated RESPIRE project (NE/W002264/1), as part of the SPF Clean Air Programme.

Polysaccharide BAP1 of Bifidobacterium adolescentis CCDM 368 is a biologically active molecule with immunomodulatory properties

Bifidobacteria are among the most common bacteria used for their probiotic properties and their impact on the maturation and function of the immune system has been well-described. Recently, scientific interest is shifting from live bacteria to defined bacteria-derived biologically active molecules. Their greatest advantage over probiotics is the defined structure and the effect independent of the viability status of the bacteria. Here, we aim to characterize Bifidobacterium adolescentis CCDM 368 surface antigens that include polysaccharides (PSs), lipoteichoic acids (LTAs), and peptidoglycan (PG). Among them, Bad368.1 PS was observed to modulate OVA-induced cytokine production in cells isolated from OVA-sensitized mice by increasing the production of Th1-related IFN-γ and inhibition of Th2-related IL-5 and IL-13 cytokines (in vitro). Moreover, Bad368.1 PS (BAP1) is efficiently engulfed and transferred between epithelial and dendritic cells. Therefore, we propose that the Bad368.1 PS (BAP1) can be used for the modulation of allergic diseases in humans. Structural studies revealed that Bad368.1 PS has an average molecular mass of approximately 9,99 × 106 Da and it consists of glucose, galactose, and rhamnose residues that are creating the following repeating unit:→2)‐β‐D‐Glcp‐1→3‐β‐L‐Rhap‐1→4‐β‐D‐Glcp‐1→3‐α‐L‐Rhap‐1→4‐β‐D‐Glcp‐1→3‐α‐D‐Galp‐(1→n

Exposure to the anti-microbial chemical triclosan disrupts keratinocyte function and skin integrity in a model of reconstructed human epidermis

Triclosan is an anti-microbial chemical incorporated into products that are applied to the skin of healthcare workers. Exposure to triclosan has previously been shown to be associated with allergic disease in humans and impact the immune responses in animal models. Additionally, studies have shown that exposure to triclosan dermally activates the NLRP3 inflammasome and disrupts the skin barrier integrity in mice. The skin is the largest organ of the body and plays an important role as a physical barrier and regulator of the immune system. Alterations in the barrier and immune regulatory functions of the skin have been demonstrated to increase the risk of sensitization and development of allergic disease. In this study, the impact of triclosan exposure on the skin barrier and keratinocyte function was investigated using a model of reconstructed human epidermis. The apical surface of reconstructed human epidermis was exposed to triclosan (0.05–0.2%) once for 6, 24, or 48 h or daily for 5 consecutive days. Exposure to triclosan increased epidermal permeability and altered the expression of genes involved in formation of the skin barrier. Additionally, exposure to triclosan altered the expression patterns of several cytokines and growth factors. Together, these results suggest that exposure to triclosan impacts skin barrier integrity and function of human keratinocytes and suggests that these alterations may impact immune regulation.

The effect of probiotics on the Californian rabbits' productivity

Against the background of an increase in allergic diseases in humans, the issue of safety and bioavailability of meat products is very relevant today. The diversity of the biochemical composition of rabbit meat and the high percentage of its nutrients' digestibility by the human body determines the value of rabbit meat. In this regard, the study of the issues of increasing the rabbits' productivity is relevant. The aim of the work was to study the effect of the Bio-Mos probiotic on the productivity of Californian rabbits.The study was conducted on the basis of the FSBEI HE SAU of the Northern Trans-Urals and on the basis of the rabbit breeding complex of CJSC APPK "Roshchinsky" of the Tyumen region. The object was male rabbits at the age of 4 months. 2 groups were formed: control (regular diet), experimental (regular diet + Bio-Mos probiotic at a dose of 2 g per kg of granulated feed). Further, the stomach contents of rabbits were evaluated; biochemical analysis of muscle tissue samples, slaughter yield, organoleptic and microbiological indicators of meat were carried out. The studies revealed a positive effect on the digestibility of nutrients (fiber, copper, and zinc), as well as on the slaughter yield (an increase of 2.5%), an increase in the amount of protein (18.30± 0.19g/100g), fat (8.5±0.64g/100g) and the energy value of meat (647.90±6.59kJ/100 g). All this indicates an increase in productivity against the background of introducing Bio-Mos into the diet due to stabilization of the digestive system.

Exposure to polycyclic aromatic hydrocarbons and serum total IgE in the Korean adults: the Third Korean National Environmental Health Survey (2015-2017).

Polycyclic aromatic hydrocarbons (PAHs) have become common pollutants with industrial development. Although the effect of exposure to PAHs on allergic disease in humans has been evaluated, evidence of an association is sparse. The association between PAH exposure and serum total immunoglobulin E (IgE) levels was evaluated in Korean adults. In total, this study included 3,269 participants in the Third Korean National Environmental Health Survey (2015-2017). Four urinary PAH metabolites were used to assessed exposure to PAHs: 1-hydroxypyrene, 1-hydroxyphenanthrene, 2-naphthol, and 2-hydroxyfluorene. The analyses were performed on 3 cutoff levels (100 IU/mL, 114 IU/mL, and 150 IU/mL) set as the total IgE elevation. Prevalence of total IgE elevation by PAH exposure group and general characteristics (age, sex, BMI, smoking, alcohol drinking, and occupation) were analyzed using the Rao-Scott χ2 test. Multiple logistic regression analyses were conducted to calculate adjusted odds ratios (ORs) for total IgE elevation by PAH exposure groups. Total IgE elevation differed significantly by age, sex, smoking status, alcohol drinking status, and occupation. For 2-hydroxyfluorene, the fourth quartile showed a significant association with IgE elevation compared to the first quartile in the analyses of cutoff-level 100 IU/mL (OR: 1.372, 95% confidence interval [CI]: 1.007-1.869) and 114 IU/mL (OR: 1.643, 95% CI: 1.167-2.312). In the analysis of cutoff-level 150 IU/mL, the adjusted ORs of the third and fourth quartile of 2-hydroxyfluorene were significantly higher than the first quartile (3rd quartile: OR: 1.478, 95% CI: 1.034-2.113; 4th quartile: OR: 1.715, 95% CI: 1.161-2.534). However, there were no significant positive associations for the other metabolites. This study implied that PAHs exposure is associated with total IgE elevation in Korean adults. More research is needed to confirm the effect of exposure to PAHs on serum IgE and allergic diseases.

Recent Advances in Allergen-Specific Immunotherapy in Humans: A Systematic Review.

Allergen-specific immunotherapy (AIT) is presumed to modulate the natural course of allergic disease by inducing immune tolerance. However, conventional AITs, such as subcutaneous immunotherapy and sublingual immunotherapy, require long treatment durations and often provoke local or systemic hypersensitivity reactions. Therefore, only <5% of allergy patients receive AIT as second-line therapy. Novel administration routes, such as intralymphatic, intradermal and epicutaneous immunotherapies, and synthetic recombinant allergen preparations have been evaluated to overcome these limitations. We will review the updated views of diverse AIT methods, and discuss the limitations and opportunities of the AITs for the treatment of allergic diseases in humans.

Research Progress in Regulation of Allergic Diseases by Short-Chain Fatty Acids

Gut microbiota-derived metabolites play vital roles in the regulation of host-gut microbiota mutualism, gut homeostasis and the pathogenesis of multiple human diseases. Fermentation of indigestible dietary fibers by gut microbiota produces a variety of short-chain fatty acids (SCFAs) consisting mainly of acetate, propionate and butyrate. Despite high concentrations of SCFAs in the gut, it has been reported in a large number of studies that SCFAs are involved in the onset and development of multiple diseases, including colitis, diabetes mellitus, hepatic steatosis, and obesity. Recent studies including our work found that SCFAs regulates allergic immune reactions and the pathogenesis of allergic diseases via their action on allergic effector immune cells, including T helper 2 (Th2) cells, type 2 innate lymphoid cells (ILC2), eosinophils, mast cells and basophils. Herein, we reviewed the association of SCFAs with human allergic diseases, their role in regulating the animal model of allergic diseases and the effects of different SCFAs in regulating the functions of allergic effectors cells and the underlying mechanisms, aiming to provide research clues for in-depth investigation in the role played by SCFAs in regulating various allergic diseases.

Microbiota and human allergic diseases: the company we keep.

Environmental, maternal and early life microbial/immune networks program human developmental trajectories and health outcomes and strongly modify allergic disease risk. The effects of environmental microbiota are illustrated by the 'farm effect' (the protection against asthma and allergy conferred by growing up on a traditional farm) and other natural experiments in populations exposed to microbe-rich environments. The role of gut microbiome maturation in the asthma/allergy trajectory is demonstrated by the most recent farm studies, which identified microbial metabolites specifically associated with asthma protection, and studies in other cohorts, which defined dynamic microbial community profiles associated with allergic disease phenotypes. Current and future studies in germ-free mice associated with gut microbiota from human disease states are providing novel mechanistic insights into the role of microbiota in shaping immune function and allergic disease susceptibility.

Environmental allergens trigger type 2 inflammation through ripoptosome activation

Environmental allergens including fungi, insects and mites trigger type 2 immunity; however, the innate sensing mechanisms and initial signaling events remain unclear. Herein, we demonstrate that allergens trigger RIPK1-caspase 8 ripoptosome activation in epithelial cells. The active caspase 8 subsequently engages caspases 3 and 7, which directly mediate intracellular maturation and release of IL-33, a pro-atopy, innate immunity, alarmin cytokine. Mature IL-33 maintained functional interaction with the cognate ST2 receptor and elicited potent pro-atopy inflammatory activity in vitro and in vivo. Inhibiting caspase 8 pharmacologically and deleting murine Il33 and Casp8 each attenuated allergic inflammation in vivo. Clinical data substantiated ripoptosome activation and IL-33 maturation as likely contributors to human allergic inflammation. Our findings reveal an epithelial barrier, allergen-sensing mechanism that converges on the ripoptosome as an intracellular molecular signaling platform triggering type 2 innate immune responses. These findings have significant implications for understanding and treating human allergic diseases.

Protection against allergies: Microbes, immunity, and the farming effect

The prevalence of asthma and other allergic diseases has rapidly increased in "Westernized" countries over recent decades. This rapid increase suggests the involvement of environmental factors, behavioral changes or lifestyle, rather than genetic drift. It has become increasingly clear that the microbiome plays a key role in educating the host immune system and, thus, regulation of disease susceptibility. This review will focus on recent advances uncovering immunological and microbial mechanisms that protect against allergies, in particular, within the context of a farming environment. A whole body of epidemiological data disclosed the nature of the protective exposures in a farm. Current evidence points toward an important role of the host microbiome in setting an immunological equilibrium that determines progression toward, or protection against allergic diseases. Conclusive mechanistic insights on how microbial exposures prevent from developing allergic diseases in humans are still lacking but findings from experimental models reveal plausible immunological mechanisms. Gathering further knowledge on these mechanisms and confirming their relevance in humans is of great importance to develop preventive strategies for children at risk of developing allergies.