title: Identification of major antigenic peptide of filarial glutathione-S-transferase.
abstract: In our earlier report, a 26kDa Setaria cervi glutathione-S-transferase showed significant protection (82%) in jirds infected with L3 larvae of Brugia malayi. In the present study we have identified the major antigenic epitopes in ScGST. Carboxypeptidase B has been used to digest the ScGST in to smaller fragments. The digested products were separated as four protein bands on SDS-PAGE. The smallest fragment of 6kDa (P4) from ScGST was identified as major antigenic epitope because of its significant reactivity with jird anti ScGST sera and human filarial sera in immunoblotting. The MALDI-LC/MS sequencing of ScGST P4 peptide ((5)KLTYFSIRGRGLAEPIRL(20), (22)KVPDDQQFLDDLISR(36) and (47)VFHFGQGPHHGPPR(62)) suggested that this protein band has a fragment of 5-62 residues long that matched with the N-terminal end of filarial GST. The antigenicity plot of ScGST was compared with BmGST model and both exhibited three immunogenic peaks within the first 60 residues towards N-terminal. In BmGST the N-terminal region was also detected with N-glycosylation signal peptide NAS adding to its high immunogenic property. Further, P4 showed strong reactivity with IgG1 and IL-4 response in endemic normal sera suggested its role in Th2 response which in turn is correlated with antibody dependent cell mediated cytotoxicity. Thus taking these results into account we propose 5-62 residues long N-terminal peptide of GST as a potential target for further vaccination studies against filarial infection.
authors.affiliation: Department of Biochemistry, Banaras Hindu University, Varanasi 221005, India.
authors.forename: Marshleen
authors.initials: M
authors.lastname: Yadav
authors.forename: Eva
authors.initials: E
authors.lastname: Liebau
authors.forename: Chandana
authors.initials: C
authors.lastname: Haldar
authors.forename: Sushma
authors.initials: S
authors.lastname: Rathaur
mesh_terms: D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D018604:Epitope Mapping; D005368:Filariasis / Q000276:immunology / Q000517:prevention & control* / Q000662:veterinary*; D005370:Filarioidea / Q000201:enzymology* / Q000276:immunology*; D005849:Gerbillinae; D005982:Glutathione Transferase / Q000276:immunology*; D006801:Humans; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000909:Antibodies, Helminth; D005982:Glutathione Transferase