| Project/Area Number |
11671150
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General surgery
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| Research Institution | Chiba University |
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Principal Investigator |
TAKIGUCHI Nobuhiro (2000) Chiba University, School of Medicine, Assistant, 医学部, 助手 (00261917)
若月 一雄 (1999) 千葉大学, 医学部・附属病院, 助手 (60302543)
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| Co-Investigator(Kenkyū-buntansha) |
ODA Kenji Chiba University, University Hospital, Assistant, 医学部・附属病院, 助手 (90282483)
YASUTOMI Jun Chiba University, University Hospital, Assistant, 医学部・附属病院, 助手 (30323421)
滝口 伸浩 千葉大学, 医学部・附属病院, 助手 (00261917)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2000: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1999: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | Human Monoclonal Antibody / cancer associated antigen / cDNA cloning / colon cancer / AgSK1 / specific immunotherapy / cytotoxic T lymphocyte / peptide vaccine / 細胞障害性T細胞(CTL) / ペプチド抗原 / 抗イディオタイプ抗体 / 細胞傷害性T細胞(CTL) / 癌ワクチン |
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| Research Abstract |
SK1, a human IgM monoclonal antibody recognizes the antigen, termed AgSK1, which was shown to be preferentially expressed by human adenocarcinomas, particularly human gastrointestinal malignancies (Bt.J.Camcer 78, 1313-22, 1998). Recently, we screened a cDNA expression library constructed using mRNA from a colon carcinoma cell line HT29 and isolated a partial cDNA clone designated AgSK1-2HT ( Tissue Antigens 55, 157-61, 2000). This clone consisted of an amino terminal open reading frame of 54 amino acids and the carboxyl terminal 20 amino acids of this coding region contained the antigenic epitope recognized by SK1. In order to show additional evidence for our claim that the identified peptide was the main antigenic epitope recognized by SK1, we tried to determine a full length cDNA sequence of this clone. Searching the nonredundant and EST database using the BLAST program, we found a clone DJ1129D05 that contained a highly homologous fragment to the coding region of AgSK1-2HT.According to the sequence of this clone, we PCR amplified several overlapping fragments using mRNA from a human colon carcinoma cell line LoVo and directly sequenced them. Assembly of these sequences yielded the entire coding sequence of AgSK1-2HT that seemed have 387 nucleotides encoding 129 amino acid peptide ( NCBI Gen Bank accession number AF316855 and AAK15476). We also found that this clone consisted of 9-mer and 10-mer peptide which carried the strong allele specific anchor residues to HLA-A2402 and HLA-A0201 respectively. We are now trying to induce CTL using these peptides in vitro. Although we should further analyze these peptides, AgSK1-2HT may be utilized as a useful cancer vaccine.
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