A Magneto-Nanosensor Immunoassay for Sensitive Detection of Aspergillus Fumigatus Allergen Asp f 1

ABSTRACT Allergic bronchopulmonary aspergillosis (ABPA) is an immunologically complex allergic disorder caused by the fungal pathogen Aspergillus fumigatus. Elevated levels of total immunoglobulin E (IgE), specific IgE, and IgG antibodies in sera are important immunodiagnostic criteria for ABPA. International reference standards or standardized immunodiagnostic assays are not available due to a lack of well-defined diagnostic antigens. The present study was carried out to identify and evaluate the immunodiagnostic relevance of synthetic epitopic peptides of Asp f 1, a major allergen, antigen, or cytotoxin of A. fumigatus. Five overlapping peptides were synthesized from the N terminus of Asp f 1, one of the potential immunodominant regions predicted by algorithmic programs. The 11-amino-acid synthetic peptide (P1) significantly inhibited both IgG binding (89.10% ± 4.45%) and IgE binding (77.32% ± 3.38%) of the standardized diagnostic antigen (SDA) (a well-defined pool of diagnostically relevant allergens and antigens of A. fumigatus). With a panel of sera of ABPA patients, allergic patients with skin test negativity to A. fumigatus, and healthy individuals, P1 showed a higher diagnostic efficiency than SDA (specific IgG, 100%; specific IgE, 98.3%). The diagnostic efficiency of P1 could be attributed to the presence of homologous epitopes in various immunodominant allergens or antigens of A. fumigatus. The ability of P1 to induce histamine release from sensitized mast cells and a Th2 type of cytokine profile in peripheral blood mononuclear cells of ABPA patients suggests its potential for use in intradermal testing. P1 could be further explored for development of a standardized, specific, and sensitive immunodiagnostic test for aspergillosis.

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Vaccinations with Recombinant Variants of Aspergillus fumigatus Allergen Asp f 3 Protect Mice against Invasive Aspergillosis

Infection and Immunity ◽

10.1128/iai.00815-06 ◽

2006 ◽

Vol 74 (9) ◽

pp. 5075-5084 ◽

Cited By ~ 57

Author(s):

James I. Ito ◽

Joseph M. Lyons ◽

Teresa B. Hong ◽

Daniel Tamae ◽

Yi-Kuang Liu ◽

...

Keyword(s):

Invasive Aspergillosis ◽

Aspergillus Fumigatus ◽

Mononuclear Cells ◽

Protein Vaccine ◽

Novel Approach ◽

In Vivo Testing ◽

Asp F 1 ◽

Fungal Allergen ◽

Subcutaneous Immunization

ABSTRACT A vaccine that effectively protects immunocompromised patients against invasive aspergillosis is a novel approach to a universally fatal disease. Here we present a rationale for selection and in vivo testing of potential protein vaccine candidates, based on the modification of an immunodominant fungal allergen for which we demonstrate immunoprotective properties. Pulmonary exposure to viable Aspergillus fumigatus conidia as well as vaccination with crude hyphal extracts protects corticosteroid-immunosuppressed mice against invasive aspergillosis (J. I. Ito and J. M. Lyons, J. Infect. Dis. 186:869-871, 2002). Sera from the latter animals contain antibodies with numerous and diverse antigen specificities, whereas sera from conidium-exposed mice contain antibodies predominantly against allergen Asp f 3 (and some against Asp f 1), as identified by mass spectrometry. Subcutaneous immunization with recombinant Asp f 3 (rAsp f 3) but not with Asp f 1 was protective. The lungs of Asp f 3-vaccinated survivors were free of hyphae and showed only a patchy low-density infiltrate of mononuclear cells. In contrast, the nonimmunized animals died with invasive hyphal elements and a compact peribronchial infiltrate of predominately polymorphonuclear leukocytes. Three truncated versions of rAsp f 3, spanning amino acid residues 15 to 168 [rAsp f 3(15-168)], 1 to 142, and 15 to 142 and lacking the known bipartite sequence required for IgE binding, were also shown to be protective. Remarkably, vaccination with either rAsp f 3(1-142) or rAsp f 3(15-168) drastically diminished the production of antigen-specific antibodies compared to vaccination with the full-length rAsp f 3(1-168) or the double-truncated rAsp f 3(15-142) version. Our findings point to a possible mechanism in which Asp f 3 vaccination induces a cellular immune response that upon infection results in the activation of lymphocytes that in turn enhances and/or restores the function of corticosteroid-suppressed macrophages to clear fungal elements in the lungs.

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Optimal Aspergillus fumigatus and Asp f 1 serum IgG cut-offs for the diagnosis of allergic bronchopulmonary aspergillosis

Allergology International ◽

10.1016/j.alit.2020.07.006 ◽

2021 ◽

Vol 70 (1) ◽

pp. 74-80

Author(s):

Yuto Hamada ◽

Yuma Fukutomi ◽

Eiji Nakatani ◽

Akemi Saito ◽

Kentaro Watai ◽

...

Keyword(s):

Aspergillus Fumigatus ◽

Allergic Bronchopulmonary Aspergillosis ◽

Serum Igg ◽

Asp F 1

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Indoor Exposure to Mould Allergens

Archives of Industrial Hygiene and Toxicology ◽

10.2478/10004-1254-62-2011-2126 ◽

2011 ◽

Vol 62 (4) ◽

pp. 371-380 ◽

Cited By ~ 12

Author(s):

Ljerka Prester

Keyword(s):

Aspergillus Fumigatus ◽

Alternaria Alternata ◽

Respiratory Diseases ◽

Polyclonal Antibodies ◽

Indoor Environments ◽

Indoor Exposure ◽

Poultry Farms ◽

Asp F 1 ◽

Immunosorbent Assays

Indoor Exposure to Mould AllergensHumid indoor environments may be colonised by allergenic filamentous microfungi (moulds),Aspergillusspp.,Penicilliumspp.,Cladosporiumspp., andAlternariaspp. in particular. Mould-induced respiratory diseases are a worldwide problem. In the last two decades, mould allergens and glucans have been used as markers of indoor exposure to moulds. Recently, mould allergens Alt a 1 (Alternaria alternata) and Asp f 1 (Aspergillus fumigatus) have been analysed in various environments (residential and occupational) with enzyme-linked immunosorbent assays, which use monoclonal or polyclonal antibodies. Household Alt a 1 and Asp f 1 levels were usually under the limit of the method detection. By contrast, higher levels of mould allergens were found in environments with high levels of bioaerosols such as poultry farms and sawmills. Data on allergen Alt a 1 and Asp f 1 levels in agricultural settings may provide information on possible colonisation of respective moulds and point out to mould-related diseases in occupants.

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Determination of Aspergillus Fumigatus Allergen 1 in Poultry Farms Using the Enzyme Immunoassay

Archives of Industrial Hygiene and Toxicology ◽

10.2478/10004-1254-61-2010-2004 ◽

2010 ◽

Vol 61 (2) ◽

pp. 167-173 ◽

Cited By ~ 6

Author(s):

Ljerka Prester ◽

Jelena Macan ◽

Kristina Matković ◽

Marija Vučemilo

Keyword(s):

Aspergillus Fumigatus ◽

Enzyme Immunoassay ◽

Limit Of Detection ◽

Major Allergen ◽

Indoor Dust ◽

Enzyme Linked Immunosorbent Assay ◽

Poultry Farms ◽

Asp F 1 ◽

Common Genus

Determination ofAspergillus FumigatusAllergen 1 in Poultry Farms Using the Enzyme ImmunoassayPoultry farms contain high levels of allergenic fungi, andAspergillusspp. is the most common genus of moulds.Aspergillus fumigatusantigens are responsible for the development of several respiratory diseases including asthma. The aim of this study was to measure the mass fraction of Asp f 1, a major allergen ofAsperillus fumigatusin 37 indoor dust samples collected from four poultry farms in a rural area of the Zagreb County (Croatia) using the enzyme-linked immunosorbent assay. More than 62 % of dust samples had detectable Asp f 1 levels (limit of detection 3.6 ng g-1). The overall mean Asp f 1 level was 17.9 ng g-1[range (3.8 to 72.4) ng g-1]. Satisfactory results were obtained for analytical within-run imprecision (6.7 %), between-run imprecision (10.5 %), and accuracy (91 % to 115 %). Microclimate parameters (air temperature, relative humidity, and velocity) were within the recommended ranges in all poultry farms. This study has shown that Asp f 1 settles on dust at poultry farms and that occupational exposure to this allergen deserves monitoring in livestock buildings.

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Immunodominant peptide epitopes of allergen, Asp f 1 from the fungus aspergillus fumigatus

Peptides ◽

10.1016/s0196-9781(98)00113-2 ◽

1998 ◽

Vol 19 (9) ◽

pp. 1469-1477 ◽

Cited By ~ 28

Author(s):

Viswanath P Kurup ◽

B Banerjee ◽

P.S Murali ◽

P.A Greenberger ◽

M Krishnan ◽

...

Keyword(s):

Aspergillus Fumigatus ◽

Peptide Epitopes ◽

Asp F 1

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Molecular allergen sensitization of Aspergillus fumigatus in allergic bronchopulmonary aspergillosis in Guangzhou, Southern China

10.21203/rs.3.rs-16458/v1 ◽

2020 ◽

Author(s):

Wenting Luo ◽

Haisheng Hu ◽

Zehong Wu ◽

Nili Wei ◽

Huimin Huang ◽

...

Keyword(s):

Aspergillus Fumigatus ◽

Southern China ◽

Allergic Bronchopulmonary Aspergillosis ◽

Strongly Correlated ◽

Guangzhou City ◽

Allergen Components ◽

Sige Level ◽

Asthma Patients ◽

Asp F 1 ◽

Fungal Allergen

Abstract Background Few studies have assessed the sensitization of mycotic allergens and Aspergillus fumigatus molecular allergen. This study aimed to investigate the connection between A.fumigatus components and mycotic allergens between allergic bronchopulmonary aspergillosis (ABPA) patients and A. fumigatus (Af)-sensitized asthma. Methods Serum Penicillium chrysogenum , Cladosporium herbarum , Mucor racemosus , Candida albicans , Alternaria alternata , Helminthosporium halodes and A.fumigatus allergen components (Asp f 1, Asp f 2, Asp f 3, Asp f 4, and Asp f 6) sIgE level were measured via ImmunoCAP assay in 18 ABPA patients and 54 Af-sensitized asthma patients in Guangzhou city. Results 94.44% of ABPA patients and 87.04% of Af-sensitized asthma patients were co-sensitization to at least one another fungal allergen. The positivity rates of C. albicans ( P < 0.05) and A.alternata ( P < 0.05) were higher in ABPA than that in Af-sensitized asthma patients. The positive rates of Asp f 1 (88.89% vs 59.26%, P < 0.05), Asp f 2 (66.67% vs 33.33%, P < 0.05), Asp f 4 (61.11% vs 33.33%, P < 0.05), and Asp f 6 (66.67% vs 14.81%, P < 0.001) in ABPA were higher than those in Af-sensitized asthma patients. Patients with ABPA had higher IgE levels of Asp f 1 ( P < 0.05), Asp f 4 ( P < 0.05) and Asp f 6 ( P < 0.001) than those of Af-sensitized asthma patients. A.fumigatus was strongly correlated with C.herbarum ( r s = 0.688) in ABPA and A.alternata ( r s = 0.692) in Af-sensitized asthma patients. Optimal scale analysis was show that ABPA was more relevant to Af-components. (Cronbach`s alpha = 90.7%) Conclusion The A.fumigatus components and it`s connection with various mycotic allergens were different in ABPA and Af-sensitized asthma patients. This findings can expected to help local doctors in the diagnosis and immunotherapy of fungal allergies.

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