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

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，作为避孕疫苗的潜在候选物。重组猪透明带蛋白（r-pZP1）是从猪透明带1（1-198）的重组融合蛋白中分离得到的。rec-pZP1反应性T细胞克隆（r-pZP1T-2）是通过对先前描述的技术进行轻微修改获得的（Shigeta, Fathman, 1981）。r-pZP1T-2是抗原特异性的，遗传上局限于H-2^d表型。r-pZP1T-2可以用r-pZP1 （198mer）和合成肽- tlk （20mer）增殖，但不能用CTY （18mer）和KRV （31mer）增殖。这些结果表明，在TLK肽（20mer）中至少存在一个T细胞表位，而在CTY或KRV中没有r-pZP1T-2的T细胞表位。采用从20个氨基酸的肽（TLK）中逐个去除氨基酸的方法合成肽。r-pZP1T-2与这些肽的增殖反应表明，65-71 YEACTKR的7mer基序似乎对rec-pZP1T-2的表位至关重要。抑制猪精子与zp相互作用的MAb-5H4的B细胞表位位于rec-pzp1的54-61号氨基酸上。因此，我们很容易推测，T细胞表位65-71的7mer基序与B细胞表位54-61的8mer基序密切相关，并有助于诱导辅助性T细胞产生B细胞对MAb-5H4表位的抗体。

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.