人端粒酶逆转录酶在肿瘤免疫治疗中的应用

寻找有效的抗原表位是近期肿瘤免疫治疗的热点．人端粒酶逆转录酶(hTERT)的免疫学特性使其成为肿瘤免疫治疗中一个吸引人的目标。从人和鼠系统获得的数据证实，细胞毒淋巴细胞(CTL)能识别hTERT特异的肽并杀死多种组织类型hTERT表达的肿瘤细胞。由于hTERT在人肿瘤组织中的广泛表达，在极少正常组织的低水平表达，临床试验已开始检验将hTERT作为肿瘤免疫治疗目标的可行程度。近期树突状细胞(DC)转运系统及其相关技术的发展和成熟提供了一个快速有效筛选抗原肽的方法。     

CLONING PROMOTER OF HUMAN SATBl GENE AND EFFECT OF ATRA AND CoCl_2 ON ITS ACTIVITY

Objective To explore the structure and activity of SATBI promoter in different cells, ATRA and CoCl2 effect on its activity. Methods Using luciferase system to assay the promoter activity of human SATB1 gene, three luciferase reporter vectors were constructed which driven by different regions of 5' untranslated sequence from human SATB1 gene, called pGL3-SP2946-luc, pGL3-SP1718-luc and pGL3-SP751-luc, and transfeted into Jurkat T,K562, U937 and Hela cells transiently using lipofectinamine, the expression activity was detected at different dosage of ATRA and CoCl2 treatment for different time course. Results The reporter gene expression from SATB1 promoter were high activity in U937 cell, moderate in Jurkat T cell, low activity in K562 cell and showed no obvious activity in Hela cell, the reporter gene expression from pGL3-SP751-luc kept on the higher lever in Jurkat T,K562 and U937 cells than the other two vectors. We also found that the repressive effect of CoCl2 on SATBI' s mRNA expression and the relative luciferase expression from pGL3-SP751~luc in U937 cell was down-regulated obviously by ATRA and CoCl2 in the concentration- and time-dependent manners. Conclusion SATB1 promoter drives gene expression with cell-specificity and its core promoter region maybe exist in the - 751 ~ - 9bp of 5' untranslated region of human SATBI gene. Combined with the experiment result we found before that SATB1 was down-regulated by ATRA in U937, the results imply that STAB1 maybe is down-regulated by ATRA and CoCl2 through its promoter in the differentiation of myeloid cell line-U937.     

肿瘤相关抗原p572肽的修饰及其功能分析

由于大多数肿瘤相关抗原是自身抗原 ,且免疫耐受通常保护个体避免发生自身免疫反应 ,所以用肿瘤相关抗原在体内刺激抗肿瘤T细胞反应时 ,免疫耐受是很难克服的障碍之一。一般认为 ,免疫耐受主要针对肿瘤的一些明显表位 ,而与HLA低亲和力的潜在表位则可能不受免疫耐受的影响。基于这一假设 ,我们设计合成了三条人端粒酶逆转录酶 (hTERT )相关的抗原肽 ,p5 72 (RLFFYRKSV )、pY5 72 (YLFFYRKSV )和pF5 72 (FLFFYRKSV ) ,p5 72是近期研究较多的一条与HLA A2 1分子低亲和力的 9肽 ,pY5 72和pF5 72是我们根据 9肽氨基酸位置特点突变合成 ,理论上它们与HLA A2 1分子的亲和力要高于p5 72 ,用T2细胞模型检测证实了这一点。DC细胞是最强的抗原提呈细胞 ,我们在试验中用DC细胞负载不同的肽在体外刺激特异性CTL的分化。     

人端粒酶逆转录酶在肿瘤免疫治疗中的应用

寻找有效的抗原表位是近期肿瘤免疫治疗的热点,人端粒酶逆转录酶(hTERT)的免疫学特性使其成为肿瘤免疫治疗中一个吸引人的目标.从人和鼠系统获得的数据证实,细胞毒淋巴细胞(CTL)能识别hTERT特异的肽并杀死多种组织类型hTERT表达的肿瘤细胞.由于hTERT在人肿瘤组织中的广泛表达,在极少正常组织的低水平表达,临床试验已开始检验将hTERT作为肿瘤免疫治疗目标的可行程度.近期树突状细胞(DC)转运系统及其相关技术的发展和成熟提供了一个快速有效筛选抗原肽的方法.     

CLONING PROMOTER OF HUMAN SATB1 GENE AND EFFECT OF ATRA AND CoCl2 ON ITS ACTIVITY

Objective To explore the structure and activity of SATB1 promoter in different cells, ATRA and CoCl2 effect on its activity. Methods Using luciferase system to assay the promoter activity of human SATB1 gene, three luciferase reporter vectors were constructed which driven by different regions of 5＇ untranslated sequence from human SATB1 gene , called pGL3-SP2946-luc , pGL3-SP1718-luc and pGL3-SP751-luc , and transfected into Jurkat T, K562, U937 and Hela cells transiently using lipofectinamine, the expression activity was detected at different dosage of A TRA and CoCl2treatment for different time course. Results The reporter gene expression from SATB1 promoter were high activity in U937 cell, moderate in Jurkat T cell, low activity in K562 cell and showed no obvious activity in Hela cell, the reporter gene expression from pGL3-SP751-1uc kept on the higher lever in Jurkat T, K562 and U937 cells than the other two vectors. We also found that the repressive effect of CoCI2 on SATBI＇s mRNA expression and the relative luciferase expression from pGL3-SP751-luc in U937 cell was down-regu- lated obviously by ATRA and CoCI2 in the concentration- and time-dependent manners. Conclusion SATB1 pro- rooter drives gene expression with cell-specificity and its core promoter region maybe exist in the - 751 - - 9bp of 5＇ untranslated region of human SATB1 gene. Combined with the experiment result we found before that SATB1 was down-regulated by ATRA in U937, the results imply that STAB1 maybe is down-regulated by ATRA and CoCl2 through its promoter in the differentiation of myeloid cell line-U937.