中文摘要：

目的：设计针对癌睾抗原PLAC1、PL2L60、MAGE-3的长肽,并检测其免疫活性。方法：利用在线数据库预测PLAC1、PL2L60、MAGE-3的CTL表位,在表位集中区域选取适当长度的长肽,化学合成并纯化后,通过体外和体内活性实验验证长肽是否具有免疫活性。结果：针对PLAC1、PL2L60、MAGE-3预测出HLA-A2限制性CTL表位并设计出7条长肽。体外自提呈、DC荷肽的ELISPOT结果和体外LDH结果显示,PLAC1 P42-64、PL2L60 P169-191、MAGE-3P101-122、MAGE-3 P152-175等4条长肽具有较好的免疫活性;体内ELISA结果显示MAGE-3 P152-175在HLA-A2.1/Kb转基因小鼠中诱导了较多的IFN-γ释放,体内LDH结果显示PLAC1 P42-64、PL2L60 P169-191、MAGE-3 P101-122、MAGE-3 P152-175等4条长肽所诱导的HLA-A2.1/Kb转基因小鼠脾淋巴细胞对靶细胞具有较高的杀伤率。结论：成功筛选鉴定出针对癌睾抗原PLAC1、PL2L60、MAGE-3的4条长肽,可用作免疫治疗疫苗。

英文摘要：

Aim： To design long peptides based on CTL epitope prediction for cancer testis antigen PLAC1,PL2L60 and MAGE-3,and to study the immune activity of the long peptides. Methods： The CTL epitope of PLAC1,PL2L60 and MAGE-3 were predicted using online databases,the long peptides with appropriate length in the epitope region was chosen,and then they were synthesized chemically and purified. Finally in vitro and in vivo activity experiments were used to verify if the long peptides had immune activity. Results： HLA-A2 restricted CTL epitopes for PLAC1,PL2L60 and MAGE-3were predicted,and 7 long peptides were designed. The autopresentation,long peptide-pulsed DC ELISPOT assay andLDH results showed that long peptides PLAC1 P42-64,PL2L60 P169-191,MAGE-3 P101-122 and MAGE-3 P152-175 had better immune activity; the ELISA results in vivo showed that MAGE-3 P152-175 induced more IFN-γ release in HLA-A2. 1/Kb transgenic mice,in vivo LDH results showed that the spleen lymphocytes from HLA-A2. 1/Kb transgenic mice induced by the long peptides PLAC1 P42-64,PL2L60 P169-191,MAGE-3 P101-122 and MAGE-3 P152-175 had higher killing rate on target cells. Conclusion： Four long peptides pointing to cancer testis antigen PLAC1,PL2L60 andMAGE-3 have been successfully identified,which could be used as immunotherapy vaccine in future.