title: Purification and immunological characterization of a 12-kDa allergen from Epicoccum purpurascens.
abstract: Exposure to Epicoccum purpurascens is implicated in respiratory allergies and asthma. Several allergens of clinical importance were identified in Epicoccum extract (EE), but only one allergen has been isolated and characterized. In the present study, a 12-kDa allergen was isolated from an Epicoccum spore-mycelial extract by concanavalin-A sepharose, reverse-phase hydrophobic and gel filtration chromatography. The purified protein was recognized as a single 12-kDa allergen on immunoblot with a serum pool of Epicoccum-sensitive patients. Of the 94 respiratory allergy patients tested intradermally, 17 showed marked positive skin reactions to EE and 12 of them reacted with the 12-kDa protein, indicating a diagnostic sensitivity of 70%. More than 80% patients' sera showed immunoglobulin E (IgE) reactivity to the purified protein in enzyme-linked immunosorbent assay and immunoblot, identifying it as a major allergen. Preincubation of pooled serum with the protein led to inhibition of IgE binding to solid-phase-bound EE (effective concentration 50%=180 ng). Twelve of the 17 serum samples showed significant basophil histamine release upon stimulation with purified protein. The protein induced significant proliferation of peripheral blood mononuclear cells of 13 patients. A high level of interleukin-4 in the culture supernatant of these cells indicated induction of a T-helper type 2 response. The purified 12-kDa protein is a clinically relevant allergen and has potential for the diagnosis and therapy of Epicoccum allergies.
authors.affiliation: Allergy and Immunology Section, Institute of Genomics and Integrative Biology (CSIR), Delhi University Campus, Delhi, India.
authors.forename: Neetu
authors.initials: N
authors.lastname: Kukreja
authors.forename: Susheela
authors.initials: S
authors.lastname: Sridhara
authors.forename: Bhanu Pratap
authors.initials: BP
authors.lastname: Singh
authors.forename: Shailendra Nath
authors.initials: SN
authors.lastname: Gaur
authors.forename: Naveen
authors.initials: N
authors.lastname: Arora
mesh_terms: D000485:Allergens / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification*; D001203:Ascomycota / Q000276:immunology*; D005656:Fungal Proteins / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification*; D006385:Hemagglutination Inhibition Tests; D006636:Histamine Release; D006801:Humans; D006967:Hypersensitivity / Q000097:blood / Q000276:immunology / Q000382:microbiology*; D007073:Immunoglobulin E / Q000097:blood; D007428:Intradermal Tests; D008213:Lymphocyte Activation; D008970:Molecular Weight; D053719:Tandem Mass Spectrometry
chemical_list: D000485:Allergens; D005656:Fungal Proteins; D007073:Immunoglobulin E