MHC class IIに提示されるヒト癌抗原の分子クローニングとそのワクチン効果

MHC II 类呈递的人类癌症抗原的分子克隆及其疫苗功效

• 批准号: 12217126
• 负责人: 佐原 弘益
• 金额: $ 2.62万
• 依托单位: Sapporo Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Priority Areas (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12217126/
• 关键词: HLA-DR8; 口腔扁平上皮癌; CD4陽性T細胞; mass spectrum解析; 腫瘍抗原; alpha-enolase

われわれは,口腔扁平上皮癌細胞株OSC-20とその癌細胞を特異的に認識するCD4陽性T細胞のラインTcOSC-20の樹立と,TcOSC-20の認識する抗原は,OSC-20のHLA-DR08032に提示されている可能性が高いことを報告してきた。本年度は,これらの自然な形で提示されている抗原ペプチドの癌細胞からの生化学的単離,同定と,TcOSC-20に認識されるCD4T細胞エピトープの解析を進めた。TcOSC-20の認識する抗原を同定するために,以下のようなストラテジーを用いた。即ち,癌細胞を大量培養し,発現する抗原を酸抽出し,得られた細胞抽出物をmass spectrumにより解析し,そのデータをもとに,ペプチドを生化学的に合成し,適当な抗原提示細胞にパルスさせた後,自家T細胞を用いて,その反応性を検討するというものである。酸抽出法に関しては,基本的にわれわれの研究室においてかつて胃癌抗原の同定に用いた方法を用いた。抽出したライセートをRP-HPLCを用いて2回にわたって精製し,各フラクションについて,TcOSC-20による反応性を検討した。この結果,第2精製におけるフラクション6には,陽性コントロールである自家癌細胞にも匹敵する,極めて高い特異的細胞傷害活性が認められた。このフラクション6をもとに解析をさらに進め,nano-LC/MS/MSによるmass spectrum解析とOWLデータベースを用いて得られたデータの検討を進めた結果,以下の異なる6種類のペプチドを同定しえた。即ち,histone H2A.1,protein kinase C inhibitor-1(C末の異なるものとして2種類),cytokeratin 14,histone H2B.1,そして,human alpha-enolaseである。このうち,histone H2A.1,protein kinase C inhibitor-1,human alpha-enolaseは既に報告されている,HLA-DR8に対する結合モチーフを有していた。これらの解析結果をもとに,各ペプチドを生化学的に合成し,マウスL細胞にヒトHLA-DR8遺伝子を移入して得た,LDR0803細胞に各ペプチドをパルスし,TcOSC-20による反応性をそれぞれ検討した。その結果,TcOSC-20はhumanalpha-enolase由来の17アミノ酸に対する反応性が有意に高いことが示唆された(Int.J.Cancer誌,論文投稿中)。これは悪性黒色腫以外で,CD4陽性T細胞が認識する癌抗原に関する初めての具体的な報告である。

Oral squamous cell carcinoma (OSC-20), oral squamous cell carcinoma (OSC-20), oral squamous cell carcinoma (OPLC), oral squamous cell carcinoma (OPLC), oral squamous cell carcinoma (OSC-20), oral squamous cell carcinoma (OPCE), oral squamous cell carcinoma (OPLC), oral squamous cell carcinoma (OSC-20), oral squamous cell carcinoma (OPCE), oral squamous cell carcinoma (OPLC), oral squamous cell carcinoma (OPCE), oral squamous cell carcinoma (O This year, the nature of the disease indicates that the antigens of cancer cells are not biochemical.According to the same criteria, TcOSC-20 is responsible for the further analysis of CD4T cell cycle. TcOSC-20 may recognize that the antigens are the same as those of the antigens, and that the following antigens will be used. That is, a large number of cancer cells were cultured, and the antigen was extracted with acid, and the cell extract was obtained. The mass spectrum extract was analyzed, and the biochemical synthesis was detected. When the antigen suggested that the cells were infected, their own T cells were used. Acid extraction method was used to determine the gastric cancer antigen in the laboratory. Pull out the RP-HPLC and use the following two times. Each one has a negative effect on sex, and each one has an anti-sexual response. The results showed that the second sperm had a high level of cytotoxic activity, which was highly sensitive to the specific cytotoxic activity. In the first half of this year, the results of the improved results were obtained by nano-LC/MS/MS mass spectrum analysis. The following results are the same as those in the following categories. That is, histone H2A.1 inhibitor-1 protein kinase C inhibitor-1, cytokeratin 14 H2B.1, human alpha-enolase animals. Histone H2A.1 protein kinase C inhibitor-1,human alpha-enolase reports that there is a significant increase in the number of reports, while the combination of the HLA-DR8 report and the report shows that there are significant differences between the two groups. According to the results of the analysis of the results, the biosynthesis of biochemistry was studied, the HLA-DR8 cells were transferred into the cells, the LDR0803 cells were analyzed, and the TcOSC-20 cells were analyzed. The results showed that TcOSC-20 humanalpha-enolase was caused by 17% acid reaction and intentional reflexivity (Int.J.Cancer journal, article contribution). In addition to the sexual black color, the CD4 sex T cell should know the specific report of cancer antigen antigen.

项目成果

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