A SOLID-PHASE ENZYME-LINKED IMMUNOASSAY (ELISPOT) FOR ASSESSMENT OF THE IMMUNOGENICITY OF A TUMOUR-DERIVED HO-1 PEPTIDE

摘要

Many evidences indicate that the inducible isoform HO-1 of the microsomal enzyme heme oxygenase is expressed at high level on various tumours and HO-1 plays a very important role for rapid tumour growth. The aim of this study was to use ELISpot assay based on IFN-gamma as analyte in order to assess the immunogenicity of a HO-1 peptide with the sequence APLLRWVL isolated from renal cell carcinoma tissue and specificity of HO-1 generated T-cell lines. High numbers of IFN-gamma secreting T-cells were observed in the presence of HO-1-derived peptide while only very low numbers were found in non-stimulated samples or in the presence of an irrelevant control peptide. In addition, stable HO-1-specific IFN-gamma release by established T-cell lines was observed after repeated stimulations. In summary, the ELISpot assay used in this study demonstrated that the tumour-derived HO-1 peptide, with the sequence APLLRWVL, is immunogenic in vitro by generation of T-cell lines with peptide-specific activity.