مطالعه‌ی اجزا‌ی آلرژی‌زا‌ی کلادوسپوریوم هرباروم با روش ایمونوبلاتینگ

Background and Objective: Numerous studies on Clasporidium herbarum antigens have shown that these antigens play a major role in produceing specific IgE in atopic individuals and exacerbate the patients’ clinical conditions like atopic dermatitis. Thus, in this study allergenic components of clasporium herbarum were investigated using immunoblotting technique. Materials and Methods: Cladosporium herbarum was cultured on Sabouraudۥs dextrose agar. The grown mycelia were harvested and ruptured by liquid nitrogen and glass beads. Samples were centrifuged and the supernatant was collected as crude extract. The crude extract was separated through sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The separated proteins were transferred to nitrocellulose filter and then soaked through atopic dermatitis patients’ sera. The responsive bands to IgE were revealed by antihuman IgE antibodies conjugated with enzyme in chromogenic substrate. Results: In SDS-PAGE, the crude extract of Cladosporium herbarum showed 16 different protein bands with molecular weight between 15.1 and 110 kDa. The bands with 15.1, 18.4, 25.1, 36.3, 45 and 54 kDa were identified as strong bands. In immunoblotting, the bands with molecular weights of 15.1, 18.4, 42 and 110 kDa showed a strong reaction with IgE sera from patients with atopic dermatitis. Conclusion: The results of this study showed that the strong bands in SDS-PAGE had the highest reaction with anti- Cladosporium herbarum IgE antibody in immunoblotting technique. Thus, we speculate the intensity of bands can affect IgE response. Like other studies we contend that Cladosporium herbarum antigen can initiate allergic reaction in atopic dermatitis patients.