| Co-Investigator(Kenkyū-buntansha) |
IWAHASHI Makoto Wakayama Medical School, 2nd Dept.of Surgery, Assistant Professor, 医学部, 助手 (70244738)
YAMAUE Hiroki Wakayama Medical School, 2nd Dept.of Surgery, Associate Professor, 医学部, 助手 (20191190)
TANIMURA Hiroshi Wakayama Medical School, 2nd Dept.of Surgery, Professor, 医学部, 教授 (10026990)
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| Research Abstract |
CMV pp65 specific CTL clone was established by limiting dilution method. In order to map the epitope peptide which was restricted by HLA-B35 and specific for CMV pp65, various kinds of truncated pp65 recombinant vaccinia virus were established by homologous recombination. Full (606)-pp65 rVac, 458-pp65 rVac, 383-pp65 rVac, 316-pp65 rVac, 225-pp65 rVac, 122-pp65 rVac and 0-pp65 rVac were established. Mapping was performed by CTL clone against the target infected by various kinds of pp65 truncated recombinant vaccinia virus. It was clarified that the epitope peptide was located from 112 amino acid (aa) to 225 aa from N terminal of pp65 sequence. Furthermore, in order to make sure that epitope peptide was processed on the target cells, a large amounts of target cells were cultured, and were washed by acid buffer (pH2) to remove the peptides from the target cell. By HPLC, this peptides solution was fractionated and applied to LCL (EBV immortalized cell).
For analysis of the cytotoxic activity by CTL clone. It was also clarified that epitope peptide was on the cell surface which binds with MHC molecules.
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