Mould allergen Alt a 1 spiked with the micronutrient retinoic acid reduces Th2 response and ameliorates Alternaria allergy in BALB/c mice.

BACKGROUND: We investigated the biological function of the mould allergen Alt a 1 as a carrier of micronutrients, such as the vitamin A metabolite retinoic acid (RA) and the influence of RA binding on its allergenicity in vitro and in vivo. METHODS: Alt a 1-RA complex formation was analyzed in silico and in vitro. PBMCs from Alternaria-allergic donors were stimulated with Alt a 1 complexed with RA (holo-Alt a 1) or empty apo-Alt a 1 and analyzed for cytokine production and CD marker expression. Serum IgE-binding and crosslinking assays to apo- and holo-protein were correlated to B-cell epitope analysis. Female BALB/c mice already sensitized to Alt a 1 were intranasally treated with apo-Alt a 1, holo-Alt a 1 or RA alone before measuring anaphylactic response, serum antibody levels, splenic cytokines and CD marker expression. RESULTS: In silico docking calculations and in vitro assays showed that the extent of RA binding depended on the higher quaternary state of Alt a 1. Holo-Alt a 1 loaded with RA reduced IL-13 released from PBMCs and CD3+CD4+CRTh2 cells. Complexing Alt a 1 to RA masked its IgE B-cell epitopes and reduced its IgE-binding capacity. In a therapeutic mouse model of Alternaria allergy nasal application of holo-Alt a 1, but not of apo-Alt a 1, significantly impeded the anaphylactic response, impaired splenic antigen-presenting cells and induced IL-10 production. CONCLUSION: Holo-Alt a 1 binding to RA was able to alleviate Th2 immunity in vitro, modulate an ongoing Th2 response and prevent anaphylactic symptoms in vivo, presenting a novel option for improving allergen-specific immunotherapy in Alternaria allergy.

Impact of Fungal Spores on Asthma Prevalence and Hospitalization.

Despite making up a significant proportion of airborne allergens, the relationship between fungal spores and asthma is not fully explored. Only 80 taxa of fungi have so far been observed to exacerbate respiratory presentations, with Cladosporium spp., Aspergillus spp., Penicillium spp., and Alternaria spp. found to comprise the predominant allergenic airborne spores. Fungal spores have been found in indoor environments, such as hospitals and housing due to poor ventilation. Meanwhile, outdoor fungal spores exhibit greater diversity, and higher abundance and have been associated with hospitalizations from acute asthma presentations. In addition, fungal spores may be the underlying, and perhaps the "missing link", factor influencing the heightened rate of asthma presentations during epidemic thunderstorm asthma events. To improve our knowledge gap on fungal spores, airborne allergen monitoring must be improved to include not only dominant allergenic fungi but also provide real-time data to accurately and quickly warn the general public. Such data will help prevent future asthma exacerbations and thus save lives. In this review, we examine the health risks of prominent allergenic fungal taxa, the factors influencing spore dispersal and distribution, and why improvements should be made to current sampling methods for public health and wellbeing.

Double-blind, randomized, placebo-controlled trial of allergen-specific immunotherapy with the major allergen Alt a 1.

BACKGROUND: There have been few studies conducted on the efficacy and safety of specific immunotherapy with allergen extracts of fungi compared with other allergen extracts, and there are no data on the major allergen Alt a 1 of the fungus Alternaria alternata. OBJECTIVES: We sought to evaluate the efficacy and safety of subcutaneous immunotherapy with 2 different doses of Alt a 1 in patients with rhinoconjunctivitis caused by sensitization to A alternata. METHOD: We performed a multicenter, randomized, double-blind, placebo-controlled trial with Alt a 1 administered subcutaneously in patients with allergic rhinoconjunctivitis with or without controlled asthma aged 12 to 65 years. Three groups were included: the placebo group and active groups receiving 0.2 or 0.37 µg of Alt a 1 per dose. The main end point was the combined symptom and medication score. Secondary end points were cutaneous reactivity and serum IgE and IgG4 levels to Alt a 1. Recorded adverse reactions were graded according to World Allergy Organization criteria. RESULTS: There were significant reductions in the combined symptom and medication score for the 0.37-µg dose of Alt a 1 compared with placebo at 12 months of treatment. Reduced cutaneous reactivity and IgE levels, together with increased IgG4 levels, were demonstrated for the 2 active groups versus the placebo group. A similar safety profile was found for both active groups compared with the placebo group. No serious adverse drug reactions were reported. CONCLUSION: Immunotherapy with Alt a 1 was efficacious and safe, reducing the symptoms and medication consumption associated with rhinoconjunctivitis after only 1 year of treatment. The clinical benefits were associated with reduced skin reactivity and specific IgE levels and increased IgG4 levels.

Temporal variability in the allergenicity of airborne Alternaria spores.

The concentration of fungal spores in the air is traditionally considered as a proxy of allergen exposure. However, in vitro experiments have shown that the allergenicity of Alternaria spores varies depending on ecophysiological and developmental factors. Despite the potential clinical significance of these findings, it has never been verified in outdoor environments. This study, therefore, aims to investigate variability in the amount of the major allergen (Alt a 1) released from Alternaria spores in outdoor air. During the 3-year monitoring study (2014-2016), the median seasonal allergenicity of Alternaria spores exceeded 8.6 × 10-3 pg Alt a 1/spore. The most allergenic spores were collected during the driest and the most polluted season (with respect to seasonal concentrations of ozone, sulphur dioxide, and particulate matter). Within the season, daily spore allergenicity ranged from 2.4 to 34.7 × 10-3 pg Alt a 1/spore (5th-95th percentile). No repeatable effects of weather and pollution on short-term variations in Alternaria spore allergenicity were found. However, during the episodes when high-potency spores were recorded, the air masses arrived from eastern directions. Contrary, the spores with the lowest allergenicity were related to western winds. This suggests that factors such as source area (habitat types) and species diversity could be responsible for the varying exposure to Alternaria allergens. Our findings show that high and low-potency spores are recorded in the air; therefore, the airborne concentrations of fungal spores alone may not be sufficient to provide allergy sufferers and healthcare professionals with information about allergen exposure.

Allergic fungal rhinosinusitis.

Allergic fungal rhinosinusitis (AFRS) is a subset of chronic rhinosinusitis with nasal polyps (CRSwNP) characterized by antifungal IgE sensitivity, eosinophil-rich mucus (ie, allergic mucin), and characteristic computed tomographic and magnetic resonance imaging findings in paranasal sinuses. AFRS develops in immunocompetent patients, with occurrence influenced by climate, geography, and several identified host factors. Molecular pathways and immune responses driving AFRS are still being delineated, but prominent adaptive and more recently recognized innate type 2 immune responses are important, many similar to those established in patients with other forms of CRSwNP. It is unclear whether AFRS represents merely a more extreme expression of pathways important in patients with CRSwNP or whether there are other disordered immune responses that would define a distinct endotype or endotypes. Although AFRS and allergic bronchopulmonary aspergillosis share some analogous immune mechanisms, the 2 conditions do not occur commonly in the same patient. Treatment of AFRS almost always requires surgical debridement of the involved sinuses. Oral corticosteroids decrease recurrence after surgery, but other adjunctive pharmacologic agents, including topical and oral antifungal agents, do not have a firm evidence basis for use. There is good rationale for use of biologic agents that target eosinophilic inflammation or other type 2 responses, but studies in patients with AFRS are required.

[Allergic bronchopulmonary aspergillosis in patients with asthma: Results of a prospective study]. / Allergicheskii bronkholegochnyi aspergillez u bol'nykh bronkhial'noi astmoi: rezul'taty prospektivnogo issledovaniia.

AIM: To estimate the frequency of fungal sensitization and the incidence of allergic bronchopulmonary aspergillosis (ABPA) in asthmatic patients. SUBJECTS AND METHODS: A total of 140 asthmatic patients were examined. They underwent allergologic (skin tests for fungal allergens, estimation of total and fungal allergen-specific IgE levels) and mycological (microscopy and inoculation of respiratory biosubstrates) examinations. Chest computed tomography, when indicated, was done. A group of patients with ABPA and that of patients with severe asthma and fungal sensitization were identified. RESULTS: The frequency of fungal sensitization in asthmatic patients was 36%; the main allergenic fungi were Aspergillus and Alternaria. The incidence of ABPA was as high as 4% in the patients with asthma and 11% in those with severe asthma and fungal sensitization. CONCLUSION: The given current diagnostic criteria will assist practitioners to identify ABPA, to prevent its progression, and to initiate specific anti-inflammatory and antifungal therapy in due time.

Th2 and Th9 responses in patients with chronic mucocutaneous candidiasis and hyper-IgE syndrome.

BACKGROUND: STAT1 mutations cause chronic mucocutaneous candidiasis (CMC), while STAT3 mutations cause hyper-IgE syndrome (HIES). CMC and HIES patients have T helper (Th) 17 defects suffering from mucosal Candida infections, but only patients with HIES show an allergic phenotype with eczema, eosinophilia and high IgE levels. OBJECTIVE: We investigated whether differential Th2 and Th9 responses may explain the clinical differences. METHODS: Peripheral blood mononuclear cells of patients with CMC (n = 4), patients with HIES (n = 4), patients with atopic dermatitis (n = 4) and healthy volunteers (n = 13) were stimulated with Candida and Staphylococcus aureus, with and without IL-4. The cytokines IL-5, IL-13, IL-9, IL-17 and TGFß and regulatory T cells were measured in cell culture supernatants by ELISA or flow cytometry, respectively. RESULTS: Peripheral blood mononuclear cells of patients with CMC showed a significantly impaired production of the Th2 cytokines IL-5 and IL-13, especially in the presence of IL-4. Moreover, IL-9 production was significantly lower in patients with CMC compared to healthy controls. In contrast, patients with HIES and patients with AD showed normal IL-5 and IL-13 production, while IL-9 production was significantly lower in patients with HIES compared to healthy controls. Although TGFß was involved in the IL-4-induced IL-9 production, TGFß levels and the frequency of regulatory T cells did not differ between patients with HIES and controls. Flow cytometry analysis demonstrated an IL-9+ IL-17+ CD4+ subset in healthy controls after stimulation with Candida which was less present in patients with HIES. CONCLUSION: Patients with CMC have a general Th defect including Th2 and Th9, while patients with HIES have normal Th2 cytokines. These differences are in line with their clinical presentation. Surprisingly, the allergic cytokine IL-9 was deficient in both HIES and CMC, suggesting a Th-17-derived origin.

Fungal compositions and diversities on indoor surfaces with visible mold growths in residential buildings in the Seoul Capital Area of South Korea.

Indoor visible mold growths are known to be associated with allergies and respiratory illnesses. However, a question remains of their compositions and diversities. Using swab sampling and high-throughput DNA sequencing, this study analyzed taxonomic compositions and diversities of fungi on indoor surfaces laden with visible mold growths in residential apartments in South Korea. The sequencing results showed low species diversities with Shannon indices ranging from 0.14 to 2.29 (mean = 1.11). Several allergy-related genera were detected on the same surface, where the most abundant Cladosporium with a mean relative abundance of 41% co-occurred with less abundant Aspergillus (0.094%), Rhodotorula (6.3%), Cryptococcus (3.7%), Alternaria (4.1%), and Crivellia (17%). ß diversity analyses showed significant differences in the fungal communities between enclosed balconies and other indoor areas (P < 0.05, ANOSIM), emphasizing a need to sample at multiple indoor locations when assessments are made for indoor visible mold growths. High-throughput sequencing is powerful in characterizing compositions and diversities of fungal communities. Future studies should examine the relationships between taxonomic compositions and diversities of indoor visible molds and health outcomes of allergies and respiratory illnesses in residential buildings.

Forecasting methodologies for Ganoderma spore concentration using combined statistical approaches and model evaluations.

High concentration levels of Ganoderma spp. spores were observed in Worcester, UK, during 2006-2010. These basidiospores are known to cause sensitization due to the allergen content and their small dimensions. This enables them to penetrate the lower part of the respiratory tract in humans. Establishment of a link between occurring symptoms of sensitization to Ganoderma spp. and other basidiospores is challenging due to lack of information regarding spore concentration in the air. Hence, aerobiological monitoring should be conducted, and if possible extended with the construction of forecast models. Daily mean concentration of allergenic Ganoderma spp. spores in the atmosphere of Worcester was measured using 7-day volumetric spore sampler through five consecutive years. The relationships between the presence of spores in the air and the weather parameters were examined. Forecast models were constructed for Ganoderma spp. spores using advanced statistical techniques, i.e. multivariate regression trees and artificial neural networks. Dew point temperature along with maximum temperature was the most important factor influencing the presence of spores in the air of Worcester. Based on these two major factors and several others of lesser importance, thresholds for certain levels of fungal spore concentration, i.e. low (0-49 s m(-3)), moderate (50-99 s m(-3)), high (100-149 s m(-3)) and very high (150 < n s m(-3)), could be designated. Despite some deviation in results obtained by artificial neural networks, authors have achieved a forecasting model, which was accurate (correlation between observed and predicted values varied from r s = 0.57 to r s = 0.68).

Analysis of asthma patients for cryptococcal seroreactivity in an urban German area.

Cryptococcus neoformans is an opportunistic fungal pathogen that causes lung inflammation and meningoencephalitis in immunocompromised patients but is also able to asymptomatically infect immunocompetent individuals. C. neoformans is found ubiquitously especially in urban areas where it is spread by pigeons, and fungal exposure may predispose for asthma development already at an early age, as soon as confronted with pigeon droppings. In the study presented here, we investigated the presence of specific immunoglobulin G (IgG) against C. neoformans in sera from patients suffering from asthma in comparison to a healthy control cohort, accrued from the Leipzig Research Centre for Civilization Diseases (LIFE). For serological analysis we developed a flow cytometry (FACS) based assay specific for an acapsular strain of C. neoformans to comprehensively analyze different cryptococcal serotypes. Compared with the non-asthmatic cohort, asthmatics exhibited, as expected, an elevated level of total serum immunoglobulin E (IgE), whereas the IgG seroreactivity against C. neoformans was not significantly different among the two groups (P = .118). Nevertheless, there was a trend toward increased Cryptococcus-specific IgG antibodies in the serum of asthmatics. Additionally, in male asthmatics an increased IgG-mediated seroreactivity compared to female asthmatics was found. This points to a higher prevalence of subclinical C. neoformans infection in male asthmatics and may support the hypothesis of C. neoformans as a risk factor for the development of asthma in urban areas.

Going over Fungal Allergy: Alternaria alternata and Its Allergens.

Fungal allergy is the third most frequent cause of respiratory pathologies and the most related to a poor prognosis of asthma. The genera Alternaria and Cladosporium are the most frequently associated with allergic respiratory diseases, with Alternaria being the one with the highest prevalence of sensitization. Alternaria alternata is an outdoor fungus whose spores disseminate in warm and dry air, reaching peak levels in temperate summers. Alternaria can also be found in damp and insufficiently ventilated houses, causing what is known as sick building syndrome. Thus, exposure to fungal allergens can occur outdoors and indoors. However, not only spores but also fungal fragments contain detectable amounts of allergens and may function as aeroallergenic sources. Allergenic extracts of Alternaria hyphae and spores are still in use for the diagnosis and treatment of allergic diseases but are variable and insufficiently standardised, as they are often a random mixture of allergenic ingredients and casual impurities. Thus, diagnosis of fungal allergy has been difficult, and knowledge about new fungal allergens is stuck. The number of allergens described in Fungi remains almost constant while new allergens are being found in the Plantae and Animalia kingdoms. Given Alt a 1 is not the unique Alternaria allergen eliciting allergy symptoms, component-resolved diagnosis strategies should be applied to diagnose fungal allergy. To date, twelve A. alternata allergens are accepted in the WHO/IUIS Allergen Nomenclature Subcommittee, many of them are enzymes: Alt a 4 (disulfide isomerase), Alt a 6 (enolase), Alt a 8 (mannitol de-hydrogenase), Alt a 10 (aldehyde dehydrogenase), Alt a 13 (glutathione-S-transferase) and Alt a MnSOD (Mn superoxide dismutase), and others have structural and regulatory functions such as Alt a 5 and Alt a 12, Alt a 3, Alt a 7. The function of Alt a 1 and Alt a 9 remains unknown. Other four allergens are included in other medical databases (e.g., Allergome): Alt a NTF2, Alt a TCTP, and Alt a 70 kDa. Despite Alt a 1 being the A. alternata major allergen, other allergens, such as enolase, Alt a 6 or MnSOD, Alt a 14 have been suggested to be included in the diagnosis panel of fungal allergy.

Survey of Sensitization to Common Fungi in an Allergic Dog Population: The Need for Further Focus on Sensitization and Allergy to Fungi in Veterinary Medicine.

Most fungal species are commensals and non-pathogenic to plants, humans, or animals. However, several species of the Alternaria, Aspergillus, Trichophyton, and Microsporum genera are common causes of disease, even for immunocompetent individuals. Besides mucosal damage, fungi may contribute to a skin barrier impairment, favoring sensitization and allergy development. A total of 68 allergic dogs were selected from a veterinary dermatology and allergy outpatient consultation for conditions related to both Malassezia overgrowth and other fungal complications. The allergy diagnosis was made through anamnesis and current clinical criteria, with the involved allergenic species being identified by intradermal tests (IDTs) and allergen-specific immunoglobulin E (sIgE) determination in serum. Dermatophagoides farinae, Dactylis glomerata, and Malassezia pachydermatis showed as the higher sensitization species from house dust mites, grass pollen, and fungi, respectively. Significant correlations at p < 0.05 were found between sensitization to Dactylis glomerata and Phleum pratense grass pollens, Dermatophagoides farinae and Dermatophagoides pteronyssinus, Acarus siro, Tyrophagus putrescentiae, and Lepidoglyphus destructor dust/storage mites, and between fungi like Aspergillus mix and Penicillium or Alternaria alternata. A significant correlation was also found between sensitization to the Aspergillus mix and D. farinae, D. pteronyssinus, or A. siro. Rather severe dermatitis was observed when a positive IDT to Malassezia pachydermatis was found, regardless of the detection of circulating sIgE, allowing us to consider the usefulness of both the IDT and the sIgE for a systematic diagnosis of allergy to fungi.

Problems Encountered Using Fungal Extracts as Test Solutions for Fungal Allergy Diagnosis.

Fungal allergy is a worldwide public health burden, and problems associated with a reliable allergy diagnosis are far from being solved. Especially, the lack of high-quality standardized fungal extracts contributes to the underdiagnosis of fungal allergy. Compared to the manufacturing processes of extracts from other allergen sources, the processes used to manufacture extracts from fungi show the highest variability. The reasons for the high variability are manifold as the starting material, the growth conditions, the protein extraction methods, and the storage conditions all have an influence on the presence and quantity of individual allergens. Despite the vast variety of studies that have analyzed the impact of the different production steps on the allergenicity of fungal allergen extracts, much remains unknown. This review points to the need for further research in the field of fungal allergology, for standardization and for generally accepted guidelines on the preparation of fungal allergen extracts. In particular, the standardization of fungal extracts has been and will continue to be difficult, but it will be crucial for improving allergy diagnosis and therapy.

Effect of yeast probiotic Saccharomyces boulardii cell wall extract on Aspergillus fumigatus allergenicity in A549 cells.

Background and Purpose: Interest in probiotic use for respiratory allergies has increased. In this regard, the present study aimed to evaluate the effect of cell wall extract of Saccharomyces boulardii on Aspergillus fumigatus as an allergenic fungus and its effectiveness in reducing inflammatory cytokines in A549 cells sensitized with A. fumigatus conidia. Materials and Methods: Cell wall of S. boulardii was prepared and challenged by A. fumigatus conidia at various concentrations. Secretory protease activity was tested using the Casein method. The A. fumigatus allergen 1 (Asp f1) gene expression was calculated by quantitative real-time polymerase chain reaction (qRT-PCR). In another experiment, qRT-PCR was used to examine gene expression of interleukin 13 and interleukin 17 by A549 lung epithelial cells exposed to A. fumigatus conidia and treated with different concentrations of S. boulardii cell wall extract. Results: Saccharomyces boulardii cell wall extract significantly reduced the protease activity of A. fumigatus at concentrations of 10 and 20 mg/ml (P<0.05). The Asp f1 gene expression was significantly down-regulated in each concentration of S. boulardii cell wall extract (P<0.05). Aspergillus fumigatus conidia upregulated the expression of IL-13 and IL-17 in A549 cells, and S. boulardii cell wall extract could downregulate the expression of the mentioned cytokines at concentrations of 10 and 20 mg/ml (P<0.05). Conclusion: According to the results, it can be concluded that S. boulardii cell wall extract could be a candidate for IL-13- and IL-17-induced Aspergillus-mediated allergy and asthma therapies. Nevertheless, future studies need to be conducted on the safety of S. boulardii cell wall extract in vivo and its effects on other arms of allergic hypersensitivity.

Fungal Allergen and Mold Allergy Diagnosis: Role and Relevance of Alternaria alternata Alt a 1 Protein Family.

Alternaria is a genus of worldwide fungi found in different habitats such as soil, the atmosphere, plants or indoor environments. Alternaria species are saprobic-largely involved in the decomposition of organic material-but they can also act as animal pathogens, causing disease in humans and animals, developing infections, toxicosis and allergic diseases. A. alternata is considered one of the most important sources of fungal allergens worldwide and it is associated with severe asthma and respiratory status. Among the A. alternata allergens, Alt a 1 is the main sensitizing allergen and its usefulness in diagnosis and immunotherapy has been demonstrated. Alt a 1 seems to define a protein family that can be used to identify related pathogenic fungi in plants and fruits, and to establish taxonomic relationships between the different fungal divisions.

Assessing global fungal threats to humans.

Fungi are an integral part of the earth's biosphere. They are broadly distributed in all continents and ecosystems and play a diversity of roles. Here, I review our current understanding of fungal threats to humans and describe the major factors that contribute to various threats. Among the 140,000 or so known species out of the estimated six million fungal species on Earth, about 10% directly or indirectly threaten human health and welfare. Major threats include mushroom poisoning, fungal allergies, infections of crop plants, food contamination by mycotoxins, and mycoses in humans. A growing number of factors have been identified to impact various fungal threats, including human demographics, crop distributions, anthropogenic activities, pathogen dispersals, global climate change, and/or the applications of antifungal drugs and agricultural fungicides. However, while models have been developed for analyzing various processes of individual threats and threat managements, current data are primarily descriptive and incomplete, and there are significant obstacles to integration of the diverse factors into accurate quantitative assessments of fungal threats. With increasing technological advances and concerted efforts to track the spatial and temporal data on climate and environmental variables; mycotoxins in the feed and food supply chains; fungal population dynamics in crop fields, human and animal populations, and the environment; human population demographics; and the prevalence and severities of fungal allergies and diseases, our ability to accurately assess fungal threats will improve. Such improvements should help us develop holistic strategies to manage fungal threats in the future.

Prevalence of Aspergillus fumigatus skin positivity in adults without an apparent/known atopic disease in Uganda.

BACKGROUND: Skin prick testing (SPT) is an important investigation in the evaluation of allergy to fungal pathogens. However, the background sensitivity to fungal allergens among healthy people in Uganda is unknown. Our aim was to assess the background prevalence of Aspergillus fumigatus SPT positivity in apparently healthy adults without known atopic disease in Uganda. METHODS: For this pilot study, we recruited 50 healthy volunteers using convenience sampling, 56% of whom were health workers. We performed the SPT for A. fumigatus according to manufacturer's instructions. A wheal diameter of ⩾3 mm was considered positive. RESULTS: The prevalence of A. fumigatus skin positivity was 60% (30/50). Participants with a positive A. fumigatus SPT were significantly younger than those with a negative result [median age (years): 28 versus 35; p = 0.005]. CONCLUSION: There is a high skin positivity against A. fumigatus among non-atopic healthy Ugandan adults. There is an urgent need to establish a normal wheal cut-off value for this population. SPT alone may be an unreliable test for the diagnosis of A. fumigatus associated allergic syndromes. More studies are needed to define the prevalence of A. fumigatus skin positivity among non-atopic healthy population in Africa.

Fungi Sensitization in Spain: Importance of the Alternaria alternata Species and Its Major Allergen Alt a 1 in the Allergenicity.

Prevalence of allergy to fungi is around 3-10%. The most prevalent species involved in sensitizations are Alternaria alternata, Aspergillus fumigatus, Cladosporium herbarum, and Penicillium notatum. Our main objective was to estimate the prevalence of fungal sensitization and its variation across Spain. Following the ICH-GCP, we recruited 1156 patients from 15 allergy departments in Spain. Hospitals were selected by bioclimatic areas. Patients underwent a skin prick test (SPT) with fungi, pollens, house dust mites, and animal dander. Specific IgE to Alternaria alternata and Alt a 1 was assessed in patients with positive SPT to fungi. Of the 233 patients (20.2%) sensitized to at least one of the five fungi tested, 162 (69.5%) were sensitized to Alternaria alternata and Alt a 1, of whom 113 (69.8%) were children; 181 (77.7%) were also polysensitized to other allergens. Alternaria alternata and Alt a 1 sensitization was present in 25.4% of patients in the Continental area, 12.0% in the Mediterranean area, 7.0% in the Semidesertic area, and 2.3% in the Oceanic area. Prevalence of sensitization to the other tested sources was 63.8% to pollens, 60.5% to house dust mite, and 38.1% to animal dander. We concluded that the prevalence of fungal allergy is increasing. Fungi are still the fourth source of allergen sensitization. Alternaria alternata sensitization is the most prevalent in allergic patients to fungi. Alt a 1 is present in almost 90% of the patients sensitized to Alternaria alternata.

Changes in the Sensitization Pattern to Alternaria alternata Allergens in Patients Treated with Alt a 1 Immunotherapy.

Alternaria alternata is the most important allergenic fungus, with up to 20% of allergic patients affected. The sensitization profile of patients sensitized to A. alternata and how it changes when treated with immunotherapy is not known. Our objective is to determine the allergen recognition pattern of allergic patients to A. alternata and to study its association to the parameters studied in a clinical trial recently published. Sera of 64 patients from the clinical trial of immunotherapy with native major allergen Alt a 1 were analyzed by immunoblotting; 98. 4% of the patients recognized Alt a 1. The percentage of recognition for Alt a 3, Alt a 4, and/or Alt a 6, Alt a 7, Alt a 8, Alt a 10 and/or Alt a 15 was 1.6%, 21.9%, 12.5%, 12.5%, and 12.5% respectively. Of the 64 patients, 45 (70.3%) only recognized Alt a 1 among the allergens present in the A. alternata extract. Immunotherapy with Alt a 1 desensitizes treated patients, reducing their symptoms and medication consumption through the elimination of Alt a 1 sensitization, which is no longer present in the immunoblotting of some patients. There may be gender differences in the pattern of sensitization to A. alternata allergens, among others.

Identifying novel allergens from a common indoor mould Aspergillus ochraceus.

The increasing burden of respiratory disease is a rising concern in India. Although chronic colonisation is primarily caused by pathogenic fungi, the common environmental fungi also play an important role in developing sensitisation. This study aims to examine the allergenic potency of mycelial proteins of a common indoor fungus Aspergillus ochraceus to a selected atopic patient cohort as well as to identify the novel IgE-binding proteins through an immunoproteomic approach. 1-D and 2-D IgE specific western blot detected the IgE reactive proteins which were identified through MALDI-TOF/TOF and manual de novo peptide sequencing. The results revealed the detection of 10 cross-reactive IgE-binding proteins. Cluster analysis of 1-D immunoblot with individual patient sera identified NADP(+)-dependent glycerol dehydrogenase (GldB) homologous protein as a major allergen, which was further purified and the allergenicity was assessed. Other IgE-binding proteins showed homology with allergens like short-chain dehydrogenase, NAD-dependent mannitol dehydrogenase, and subtilisin-like serine protease. GldB purified under native conditions showed IgE reactivity amongst the selected patient cohort, which is reported for the first time in this study. The identified IgE-binding proteins can act as candidate molecules for developing hypoallergenic vaccines for designing specific immunotherapeutic techniques to fungal allergy. THE SIGNIFICANCE OF THE STUDY: Exposure to environmental fungal allergens is directly associated with promoting allergic response as well as complicating existing respiratory disease, leading to poor respiratory health. Amongst others, Aspergillus spp. contributes to the majority of the fungal derived atopic diseases. Aspergillus ochraceus is a common indoor mould in India, however, its allergenic potency was not explored till date. In this study, we establish A. ochraceus responsible to cause an allergic response to susceptible individuals and identified 10 IgE-binding proteins using an immunoproteomics approach for the first time. A. ochraceus being unsequenced, a homology-driven proteomics approach was used to identify the IgE-binding proteins which can be extended to identify proteins from other unsequenced species. The information on the IgE-binding proteins could be used as a step towards characterising them by molecular and structural methods to investigate the molecular basis of allergenicity. This will also help to enrich the existing database of allergenic proteins and pave a way towards developing therapeutic avenues.