Allergenic components of isolates ofcladosporium herbarum

Abstract

Extracts of the mycelia were prepared and their biochemical and immunological properties were examined. Two isolates, IhlI 49632 and CBS 180.50 did not give a significant yield of mycelial mat, and the mycelial extract of ATCC 38810 was only weakly immunoreactive. In IgE immunoblots of sodium dodecylsulphate polyacrylamide electrophoresis gels, the remaining three extracts showed strongly reactive bands at 14, 17,25,35,41,47 and 97 kDa hlW and IhlI 96220 had two additional bands at 53 and 71 kDa. These extracts had 8 to 10 bands in isoelectric focusing with pl values between 3.75 to 5.2, the two predominant bands being at 4.5 and 5.2. In crossed -radioimmunoelectrophoresis using human atopic sera, IMI 96220showed one dominant and three minor allergens; IMI 49630 and CBS 121.47 had one dominant and one minor allergen. In direct radioallergosorbent tests with human atopie sera and passive cutaneous anaphylaxis tests with mouse IgE, Ih4I 49630 and 96220 were more potent than CBS 121.47. These results indicate that mycelial extracts of IMI 49630 and 96220 contain a potent but complex mixture of the major allergens of C. herbanmi and are preferred sources for a reference standard for this organism. Clmiosporiiirn is reported to be the most common airborne fungus in temperate zones (Solomon 1978) and in many countries the species C. herbarirni is a major fungal cause of inhalent allergy and allergic asthma in man (Nielsson & Aas 1976, Tarlo et al. 1988. Aukrust and Borch (1979) identified two major allergenic components of this organism by extensive CRIE studies, but significant biochemical and immunological differences have been reported among different strains (Sward-Nordmo et al. 1984). In the present investigation, six isolates of C. herboriitn were studied with the aim of identifying suitable isolate for a reference standard of this organism. The isolates, IMI 49630, IMI 49632, IMI 96220, CBS 121.47, CBS 180.50 and ATCC 38810 were examined for their antigenic and allergenic components by the techniques of crossed-radioimmunoelectrophoresis (CRIE) and SDS-PAGE immunoblotting. MATERIALS AND METHODS Preparation of mycelial extractsCultures of the isolates IhlI 49630. IMI 49632, IhlI 96220, CBS 121.47, CBS 180.50 and ATCC 38810 were grown on synthetic revised tobacco medium at 22°C for 4 weeks as reported earlier (Vijay et al. 1985). Dried mold pellicles that had been homogenized and defatted with acetone were extracted with 1:20 wlv Tris-buffer, pH 7.8, containing 0.15 M NaCl for 48h at 4°C. The suspension was centrifuged at 1000 g a t 4°C for 30 min, then at 30,000 g at 4°C for 20 min. The resulting supernatant was concentrated and diafiltered with water using an Amicon YM-5 membrane. The retentate was filtered through a 0.45 pm filter and lyophilized. A reference extract (RE) of C. herbanrrn was prepared from commercial mycelial material (Pharmacia, Uppsala, Sweden) in a similar way. Biochemical assaysProtein contents of the extracts were compared by a commercial dye-binding assay (Bio-Rad Laboratories), using bo...

Allergenic and mutagenic characterization of 14 Penicillium species

“…The discrepancy in the results could be due to the type of isolate, culture, and extraction conditions. It has been shown for other fungi such as Alternaria alternata (Vijay et al, 1984) and Cladosporium herbarum (Vijay et al, 1991) that many variables influence the production of allergens in culture and hence the potency of allergen extracts.…”

The effect of meteorological factors on the daily variation of airborne fungal spores in Granada (southern Spain)

“…Alternaria and Cladosporium have been identified as the most abundant and frequent airborne allergens in several cities in the Iberian Peninsula (Infante et al 1988;González Minero et al 1993;Fernández González et al 1993;Munuera Giner and Carrión García 1995;Herrero et al 1996) and their allergenic capacity has been reported by numerous authors (Budd 1986;Vijay et al 1991;Srivastava and Wadhwani 1992;Caretta 1992;Martínez et al 1994). Since the sporulation process and spore dispersion of fungi are greatly dependent on the climatic conditions in each area, many authors have correlated meteorological parameters with the presence and prevalence of airborne spores in the atmospheres of climatically different locations.…”

Domestic laundering: Environmental audit in Glasgow with emphasis on passive indoor drying and air quality

“…In this vein, research in West Virginia [55] cautions "allergen content in fungal extracts is highly variable" [56], with variability in particular species such as Alternaria alternata, Aspergillus fumigatus and Cladosporium herbarum [57]. It also found 6% sensitivity to Acremonium strictum, where all patients in a clinic "had symptoms consistent with allergic rhinitis or asthma"; while another USA study failed to confirm Acremonium strictum as significant [58].…”