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AIM:To characterize the biochemical and immunologicalproperties of an experimental ISCOMS vaccine prepared froma novel therapeutic polypeptide based on T cell epitopes ofHBsAg,and a heptatis B-ISCOMS was prepared andinvestigated.METHODS:An immunostimulating complexes(ISCOMS)-based vaccine containing a novel therapeutic hepatits Bpolypeptide was prepared by dialysis method,and itsformation was visualized by electron microscopy andbiochemically verified by SDS-poiyacrylamide gelelectrophoresis.Amount of the peptide within ISCOMS wasdetermined by Bradford assay,and specific CTL responsewas detected by ELISPOT assay.RESULTS:Typical cage-like structures of submicroparticlewith a diameter of about 40 nm were observed by electronmicroscopy.Results from Bradford assay showed that tielevel of peptide incorporation was about 0.33 g·L~(-1).At theparalleled position close to the sixth band of the molecularweight marker(3480 kDa)a clear band was shown in SDS-PAGE analysis,indicating successful incorporation ofpolypeptide into ISCOMS.It is suggested that ISCOMSdelivery system could efficiently improve the immunogenicityof polypeptide and elicit specific immune responses in vivoby the results of ELISPOT assay,which showed that IFN-yproducing calls(specific CTL responses)were increased(spots of ISCOMS-treated group:47±5,n=3;controlgroup:5±2,n=3).CONCLUSION.- ISCOMS-based hepatitis B polypeptide vaccine is successfully constructed and it induces a higher CTL response compared with short polypeptides vaccine in vivo.
AIM: To characterize the biochemical and immunological properties of an experimental ISCOMS vaccine prepared froma novel therapeutic polypeptide based on T cell epitopes of HBsAg, and a heptatis B-ISCOMS was prepared and investigated. METHODS: An immunostimulating complexes (ISCOMS) -based vaccine containing a novel therapeutic hepatits Bpolypeptide was prepared by dialysis method, and its formation was visualized by electron microscopy and biochemically verified by SDS-poiyacrylamide gelelectrophoresis. Amount of the peptide within ISCOMS was determined by Bradford assay, and specific CTL response was detected by ELISPOT assay .RESULTS: Typical cage-like structures of submicroparticle with a diameter of about 40 nm were observed by electron microscopy. Results from Bradford assay showed that tielevel of peptide incorporation was about 0.33 g · L -1 .A the the paralleled position close to the sixth band of the molecular weight marker (3480 kDa ) a clear band was shown in SDS-PAGE analysis, indicating successful inc orporation ofpolypeptide into ISCOMS. It is suggested that ISCOMS release system could efficiently improve the immunogenicity of polypeptide and elicit specific immune responses in vivoby the results of ELISPOT assay, which showed that IFN-yproducing calls (specific CTL responses) were increased (spots of ISCOMS-treated group: 47 ± 5, n = 3; controlgroup: 5 ± 2, n = 3) .CONCLUSION.- ISCOMS-based hepatitis B polypeptide vaccine is successfully constructed and it induces a higher CTL response compared with short polypeptide vaccine in vivo.