Research for T epitope of a zona pellucida and application for vaccine

透明带T表位的研究及疫苗应用

• 批准号: 08660353
• 负责人: SHIGETA Minoru
• 金额: $ 1.47万
• 依托单位: GRANT-IN-AID FOR SCIENTIFIC RESEARCH (C) (2)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08660353/
• 关键词: porcine zona pellucida; T cell epitope; vaccine

The aima of the present study is to establish the porcine zona pellucida specific antigen reactive T cells for the analysis of T cell epitope correlated with the specific B cell epitope as a potential candidate for a contraceptive vaccine. Recombinant porcine zona pellucida protein (r-pZP1) was purified from recombinant fusion proteins for porcine zona pellucida 1 (1-198) . The rec-pZP1 reactive T cell clones (r-pZP1T-2) was obtained with slight modifications of previously described technology (Shigeta, Fathman, 1981) . The r-pZP1T-2 was antigen specific and genetically restricted to H-2^d phenotype. The r-pZP1T-2 was proliferated with r-pZP1 (198mer) and synthetic peptides-TLK (20mer) but not with CTY (18mer) nor KRV (31mer). These results show that there is at least one T cell epitope within TLK peptide (20mer) but there is no T cell epitope within CTY or KRV for the r-pZP1T-2. Peptides were synthesized in a manner to remove amino acids one by one from the 20-amino acid peptide (TLK) . Proliferative response of the r-pZP1T-2 with these peptides suggested that the 7mer motif of 65-71 YEACTKR seems to be essential for the epitope of the rec-pZP1T-2. B cell epitope for MAb-5H4 which inhibits the pig sperm-ZP interaction was determined to be present on the amino acid No.54-61 of rec-pzp1. Therefore, it is easy to speculate that 7mer motif of 65-71 for T cell epitope is closely related to 8mer motif of 54-61 for B cell epitope and help to induce helper T cells for antibody production by B cell for MAb-5H4 epitope.

本研究的目的是建立猪透明带特异性抗原反应性T细胞，用于分析与特异性B细胞表位相关的T细胞表位，作为潜在的避孕疫苗候选细胞。从猪透明质酸1（1-198）的重组融合蛋白中纯化重组猪透明质酸蛋白（r-pZP 1）。通过对先前描述的技术（Shigeta，Fathman，1981）的轻微修改获得rec-pZP 1反应性T细胞克隆（r-pZP 1 T-2）。r-pZP 1 T-2是抗原特异性的，并且遗传上限于H-2^d表型。r-pZP 1 T-2在198聚体的r-pZP 1和20聚体的合成肽TLK的作用下增殖，而在18聚体的CTY和31聚体的KRV的作用下不增殖。这些结果表明，对于r-pZP 1 T-2，在TLK肽（20聚体）内存在至少一个T细胞表位，但在CTY或KRV内不存在T细胞表位。以从20个氨基酸的肽（TLK）中逐个去除氨基酸的方式合成肽。r-pZP 1 T-2与这些肽的互补反应表明，65-71 YEACTKR的7聚体基序似乎是rec-pZP 1 T-2的表位所必需的。抑制猪精子-ZP相互作用的MA B-5 H4的B细胞表位被确定存在于rec-pzp 1的第54 -61号氨基酸上。因此，很容易推测T细胞表位的65-71的7聚体基序与B细胞表位的54-61的8聚体基序密切相关，并且有助于诱导辅助T细胞用于由B细胞产生针对MA B-5 H4表位的抗体。