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Paper Information

Journal:   JOURNAL OF ADVANCES IN MEDICAL AND BIOMEDICAL RESEARCH   SUMMER 2007 , Volume 15 , Number 59; Page(s) 55 To 64.
 
Paper: 

INVESTIGATION INTO ALLERGENIC COMPONENTS OF CLADOSPORIUM HERBARUM BY IMMUNOBLOTTING TECHNIQUE

 
 
Author(s):  HEDAYATI M.T.*, KABOLI S., HAJHEYDARI Z., SHOKOUHI T., MOHAMMADPOUR TAHAMTAN R.A.
 
* SCHOOL OF MEDICINE, MAZANDARAN UNIVERSITY OF MEDICAL SCIENCES, SARI, IRAN
 
Abstract: 

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 Sabouraud's 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 re vealed 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.

 
Keyword(s): CLADOSPORIUM HERBARUM, ATOPIC DERMATITIS, IGE, IMMUNOBLOTTING
 
References: 
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