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Identification of the reactive subunits ofAspergillus umbrosusinvolved in the antigenic response in farmer's lung

机译：Identification of the reactive subunits ofAspergillus umbrosusinvolved in the antigenic response in farmer's lung

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SummaryBackgroundFarmer's lung (FL) is the most common form of extrinsic allergic alveolitis and the fungusAspergillus umbrosushas frequently been associated with FL among Finnish farmers. IgG and IgA class antibodies againstAspergillus umbrosusare found in the serum of FL patients and healthy exposed farmers. So far the immunologically reactive subunits ofAspergillus umbrosusare unknown.ObjectivesThe purpose of the study was to identify and localize the reactive subunits ofAspergillus umbrosusthat are involved in the antigenic response in FL.MethodsThe distribution of the primary antibody on the outer wall layers of 4‐ and 11 day‐old fungiAspergillus umbrosuswas studied by pre‐embedding immunolabelling with fluorescein isothiocyanate (FITC)‐ and gold‐conjugated antibodies. The surface carbohydrates of the outer wall layers were studied using FITC‐conjugated lectins.ResultsFITC‐conjugated antibodies from FL patient sera reacted with spores (conidia), phialides and hyphae, whereas the fruiting body, cleistothecia, remained unstainable. Immunofluorescence labelling was confirmed by immunolabelling electron microscopy, and gold‐conjugated IgG antibodies were found on the outer cell walls of spinulose conidia, and in a less degree on the outer cell walls of hyphae and phialides. Only single gold particles were found scattered on the surfaces of cleistothecia. The surface carbohydrates ofAspergillus umbrosusconidia. vesicles, phialides and hyphae had a high affinity to GlcNAcβl‐ and Manαl‐/Glcαl‐ specific lectins and only GlcNAcβl‐specific lectins also reacted with the outer cell walls of cleistothecia. Blocking of the antigenic sites ofAspergillus umbrosusby preincubating antigen‐coated enzyme linked immunosorbent assay (FLISA) plates with lectins caused some inhibition, indicating that sites with the highest affinity for lectins were not always antigenic sites.ConclusionGold‐ and FITC‐conjugated antibodies were distributed mostly on the outer wall layers of conidia, hyphae and phialides ofAspergillus umbrosusand GlcN Acβl ‐linked lectins had the stronge

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