Analysis of Tumor Rejection Antigen of Murine Myeloma MOPC70A Recognized by Specific Cytotoxic T cell.

特异性细胞毒性 T 细胞识别的鼠骨髓瘤 MOPC70A 肿瘤排斥抗原的分析。

• 批准号: 11670216
• 负责人: UENAKA Akiko
• 金额: $ 1.6万
• 依托单位: OKAYAMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670216/
• 关键词: murine myeloma MOPC70A; cytotoxic T cell (CTL); antigen presenting molecule; anitgen gene; expression cloning; epitope; マウス骨髄腫MOPC-70A; 標的エピトープ

Murine myeloma MOPC70A specific cytotoxic T cell (CTL) clone E10 was established from spleen cells of tumor bearing BALB/c mouse. 2. Expression cDNA library was cloned from mRNA of MOPC70A into mammalian expression vector pMET7 and divided into pools of 100 bacterial colonies. And the total 56,000 cDNA clones were tested for their ability to stimulate TNFα production by E10 CTL after transfection into COS7 transfected H-2 gene. 3. A clone (2D2) which stimulate E10 to produce TNFα and IFNγ was cloned. 4. The sequence of 2D2 provided 1398 bp long. Sequence homology analysis revealed that 2D2 is murine apoptosis inducing factor (mAIF). 5. To identify antigenic CTL epitope encoded by the 2D2 gene, the ability of deletion mutant of 2D2 gene to stimulate E10 to produce TNFα were investigated. The results suggested that the antigenic epitope resided within the region of 737-870bp of 2D2. From analysis of the synthetic candidate peptide on the region of 737-870 bp, a nonamer peptide DLGPDVGYEA sensitized E10 CTL was found as an epitope.

从荷瘤BALB/c小鼠的脾细胞中建立了小鼠骨髓瘤MOPC 70 A特异性细胞毒性T细胞（CTL）克隆E10。2.从MOPC 70 A的mRNA中克隆表达cDNA文库到哺乳动物表达载体pMET 7中，并分成100个细菌菌落的池。将56,000个cDNA克隆转染COS 7转染的H-2基因后，检测其刺激E10 CTL产生TNFα的能力。3.克隆了一个能刺激E10产生TNFα和IFNγ的克隆（2D 2）。4. 2D 2的序列长1398 bp。序列同源性分析表明，2D 2是小鼠凋亡诱导因子（mAIF）。5.为鉴定2D 2基因编码的CTL抗原表位，研究了2D 2基因缺失突变体刺激E10产生TNFα的能力。结果表明，该抗原表位位于2D 2的737- 870 bp区域。通过对737-870 bp区域的合成候选肽的分析，发现九聚体肽DLGPDVGYEA致敏的E10 CTL作为表位。

项目成果

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