Purification of a novel perforin Inhibitor

我们将建立如何保持移植物在储存液中的功能，以及如何防止受体对移植物的急性和/或慢性排斥。但是，需要从新的角度对储液进行改进。我们正在考虑癌细胞和CTL和/或NK细胞之间的关系。一般来说，癌细胞会逃避这些免疫细胞的攻击。我们证实，癌细胞有很强的穿孔素抑制剂，并摆脱这些免疫细胞。我们有许多无蛋白培养的癌细胞系（66株），并计划从这些细胞系及其条件培养基中纯化穿孔素抑制剂。根据我们的计划，我们可以从HGC-Y1和HPC-Y3的细胞裂解液和条件培养基中纯化出强穿孔素抑制剂。n端氨基酸序列分析显示为PEPAKSA PAPKKGSKKAVTK。令人惊讶的是，这个序列与人类组蛋白H2B完全相同。事实上，小牛组蛋白具有很强的穿孔素抑制剂活性。从这些结果来看，在移植物储存液中给予组蛋白并将这种抑制剂注射给急性排斥的受体将是非常有用的，因为向正常大鼠大量给予小牛组蛋白从未在任何大鼠器官中引起任何异常的组织学发现。

We are going to establish how to keep the funtion of the graft in the storage fluid, and how to prevent the acute and/or chronic rejection of the graft from recipient. However, it is necessary to Improve the storage fluid from fresh and new stand points. We are thinking about the relationship between cancer cells and CTL and/or NK cells. In general, cancer cells escape from the attack of these immune cells. We confirm that cancer cells have strong perforin inhibitor, and get away from these immune cells. We have many protein-free cultured cancer cell lines (66 lines), and made a plan to purify the perforin inhibitor from the cell lines and their conditioned media. According to our plan, we could purified strong perforin inhibitor from cell lysate and conditioned medium of HGC-Y1 and HPC-Y3. The analysis of N-terminal amino acid sequences showed PEPAKSA PAPKKGSKKAVTK. Surprisingly, this sequence was perfectly identical to human histone H2B. Indeed, calf histone has very strong perforin inhibitor activity. From these results, the administration of histone protein in the graft storage fluid and the injection of this inhibitor to the recipient with acute rejection will be very useful, since a large giving of calf histone to normal rats never caused any abnormal histological findings in any rat organs.