title: Recombinant glutathione-S-transferase a major allergen from Alternaria alternata for clinical use in allergy patients.
abstract: Recombinant proteins are used for vaccines, therapy and diagnosis of many diseases. Biological activity of these may differ from native counterpart and needs investigation. The present study aimed to compare recombinant (r) and native (n) glutathione-S-transferase (GST) from Alternaria alternata. Glutathione-S-transferase sequence showed an ORF of 696bp encoding 26-kDa protein with N-terminus conserved domain. Secondary structure of both forms was comparable with melting temperature of 57 and 59 degrees C, respectively. rGST and nGST showed similar enzymatic activity, allergenicity and potency by ELISA inhibition. Histamine release was comparable in 14/17 patients for both the GSTs. rGST and nGST induced proliferation in PBMC at different concentration. Cell supernatant revealed higher IL-4 and IL-5 levels with low levels of IFN-gamma. In summary, recombinant and native GST demonstrated similar physio-chemical, biological and immunological properties and induced comparable cell mediated and humoral response to be used for diagnosis and specific immunotherapy for the fungal allergy cases.
authors.affiliation: Institute of Genomics and Integrative Biology, Mall Road, Delhi 110007, India.
authors.forename: Jay
authors.initials: J
authors.lastname: Shankar
authors.forename: Bhanu P
authors.initials: BP
authors.lastname: Singh
authors.forename: Shailendra N
authors.initials: SN
authors.lastname: Gaur
authors.forename: Naveen
authors.initials: N
authors.lastname: Arora
mesh_terms: D000293:Adolescent; D000328:Adult; D000485:Allergens / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D000528:Alternaria / Q000201:enzymology* / Q000235:genetics / Q000276:immunology; D016022:Case-Control Studies; D005260:Female; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D005982:Glutathione Transferase / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D006801:Humans; D007428:Intradermal Tests; D008297:Male; D008875:Middle Aged; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification; D006255:Rhinitis, Allergic, Seasonal / Q000276:immunology*
chemical_list: D000485:Allergens; D005656:Fungal Proteins; D011994:Recombinant Proteins; D005982:Glutathione Transferase