Allergy Asthma Immunol Res.  2016 Sep;8(5):428-437. 10.4168/aair.2016.8.5.428.

Orthologous Allergens and Diagnostic Utility of Major Allergen Alt a 1

• Moreno A
• Pineda F
• Alcover J
• Rodríguez D
• Palacios R
• Martínez-Naves E

• KMID: 2295152
• DOI: http://doi.org/10.4168/aair.2016.8.5.428

Abstract

PURPOSE
Hypersensitivity to fungi is associated with rhinoconjunctivitis and asthma. For some fungi, such as Alternaria alternata (A. alternata), the symptoms of asthma are persistent, increasing disease severity and the risk of fatal outcomes. There are a large number of species of fungi but knowledge of them remains limited. This, together with the difficulties in obtaining adequate standardized extracts, means that there remain significant challenges in the diagnosis and immunotherapy of allergy associated with fungi. The type of indoor fungi related to asthma/allergy varies according to geographic, climatic, and seasonal factors, making their study difficult. The aim of this study was to determine hypersensitivity to indoor fungi in a population from Cuenca, Spain.
METHODS
Thirty-five patients with symptoms compatible with rhinitis or asthma who showed clear worsening of their symptoms in their homes or workplace were included. In vivo and in vitro tests were made with a battery of fungal allergens, including the species isolated in the home or workplace.
RESULTS
Ulocladium botrytis (U. botrytis) and A. alternata were the most representative species as a source of home sensitization. These species showed very high concordance in skin tests, specific IgE, and histamine release. The allergen Alt a 1, which was recognized in all patients, was detected in A. alternata, U. botrytis, and Stemphylium botryosum (S. botryosum).
CONCLUSIONS
U. botrytis and A. alternata were the most representative species as a source of home sensitization. Alt a 1 was recognized in all patients and may be considered a non-species-specific allergen that could be used as a diagnostic source of sensitization to some species of the Pleosporaceae family.

Keyword

Alternaria alternata; Ulocladium botrytis; Alt a 1; orthologous allergens; biological standardization; pleosporaceae

MeSH Terms

Allergens*
Alternaria
Asthma
Botrytis
Diagnosis
Fatal Outcome
Fungi
Histamine Release
Humans
Hypersensitivity
Immunoglobulin E
Immunotherapy
In Vitro Techniques
Rhinitis
Seasons
Skin Tests
Spain
Allergens
Immunoglobulin E

Figure

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  Fig. 1 Home collection and identification of fungal samples.

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  Fig. 2 Allergogram of allergenic extracts of fungi vs pooled sera, high, medium and low specific IgE of selected patients under reducing and non-reducing conditions. Lane 1: Candida albicans; Lane 2: Ulocladium botrytis; Lane 3: Mucor mucedo; Lane 4: Fusarium sp.; Lane 5: Trichophyton rubrum; Lane 6: Aspergillus niger; Lane 7: Alternaria alternata; Lane 8: Stachybotrys chartarum; Lane 9: Stemphylium botryosum; Lane 10: Penicillium notatum; Lane 11: Rhizopus sp.; Lane 12: Aspergillus fumigatus; Lane 13: Botrytis sp.; Lane 14: Cladosporium herbarum.

• 

  Fig. 3 (A) Dot blot of selected extracts vs pooled sera, high, medium and low specific IgE of selected patients. (duplicate samples are shown) Row A: Candida albicans, Ulocladium botrytis, Mucor mucedo, Fusarium oxysporum, Trichophyton rubrum/Row B: Aspergillus niger, Alternaria alternata, Stachybotrys chartarum, Stemphylium botryosum, Rhizopus nigricams/Row C: Aspergillus fumigatus, Botrytis cinerea, Cladosporium herbarum, Penicillium notatum, (B1) Immunoblotting of allergenic extract of Ulocladium botrytis. (B2) Immunoblotting of allergenic extract of Alternaria alternata.

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  Fig. 4 (A) ELISA inhibition of allergenic extract of Ulocladium botrytis versus Ulocladium botrytis, Alternaria alternata, Stemphylium botryosum and (B) ELISA inhibition allergenic extract of Ulocladium botrytis versus Ulocladium botrytis, Aspergillus fumigatus and Cladosporium herbarum.

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  Fig. 5 Inhibition of immunoblot with pooled sera, high, medium and low specific IgE of selected patients under reducing and non-reducing conditions: (-) No inhibition with Alt a 1; (+) Inhibition with Alt a 1; Lane 1: Candida albicans, Lane 2: Ulocladium botrytis, Lane 3: Alternaria alternata, Lane 4: Stemphylium botryosum, Lane 5: Penicillium notatum, Lane 6: Aspergillus fumigatus, Lane 7: Cladosporium herbarum.

• 

  Fig. 6 (A) 2D electrophoresis of Ulocladium botrytis extract. (B1) immunoblotting with pooled sera of selected patients. Arrows indicate the possible orthologue of Alt a 1. (B2) immunoblotting with high level of specific IgE to Ulocladium botrytis extract. (B3) immunoblotting with medium level of specific IgE to Ulocladium botrytis extract. (B4) immunoblotting with low level of specific IgE to Ulocladium botrytis extract.

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  Fig. 7 Histamine release test. Results obtained after stimulation with Alt a 1 and Ulocladium botrytis.

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