修饰的肿瘤抗原肽CEA_(610D)疫苗的体外抗肿瘤作用  

In vitro anti-tumor activity of altered tumor-associated antigen CEA_(610D) peptide

作　　者：刘晶[1] 李翠玲[1] 顾洪涛[1] 阴海鹏[1] 朱传福[2] 温培娥[1] 张玲[1] 

机构地区：[1]山东省医学科学院基础医学研究所山东省医药卫生肿瘤免疫与中药免疫重点实验室山东省现代医用药物与技术重点实验室,山东济南250062 [2]山东省血液中心HLA研究室,山东济南250014

出　　处：《中国肿瘤生物治疗杂志》2009年第2期120-124,共5页Chinese Journal of Cancer Biotherapy

基　　金：山东省医学科学院科研项目(No.2006-25)~~

摘　　要：目的:评价修饰的CEA610D抗原肽疫苗体外抗肿瘤免疫的有效性,为其临床应用提供依据。方法:多肽固相合成法合成HLA-A2CEA修饰肽(CEA610D)、HLA-A2天然肽CEA605-613(天然肽)和HLA-A2无关肽MAGE-3(无关肽),采用同种异体混合淋巴细胞与肽共孵育的方法,诱导肽特异性细胞毒性T淋巴细胞(CTL),利用MTT法检测CTL的增殖和杀伤活性;流式细胞术观察细胞表型;RT-PCR检测细胞穿孔素的表达;ELISA法检测CTL上清中IFN-γ的水平。结果:CEA610D疫苗产生肽特异性CTL的能力最强(P<0.05),其CD8+T细胞数也高于天然肽组和无关肽组;在效靶比为40:1时,CEA610D和天然肽所诱导的CTL对CEA+HLA-A2限制性人结肠癌细胞T84的杀伤活性可达(56.7±3.73)%和(51.2±1.86)%,而3种肽特异性CTL细胞对CEA+HLA-A2人结肠癌lovo细胞杀伤活性维持在本底水平;CEA610D组的CTL所释放的杀伤相关介质穿孔素和上清中IFN-γ的水平也显著高于其余两组(P<0.01)。结论:与天然肽相比,CEA610D疫苗可打破自身表位的免疫耐受,在体外抗肿瘤免疫中更具优势。Objective ：To investigated the in vivo efficacy of altered CEA610D peptide vaccine against tumor, so as to provide a basis for its clinical use. Methods： Altered CEA peptide CEA610D, natural CEA peptide CEA605-613 and MAGE- 3 peptide were synthesized by polypeptide solid-phase synthesis system. The cytotoxicity T lymphocytes （CTLs） were induced by autologous mixed lymphocytes reaction implused with above peptides; proliferation and cytotoxicity of different CTLs were measured by MTT; phenotypes of the CTLs were detected by flow cytometry ; expression of perforin in different CTLs were assayed by RT-PCR; IFN-γ levels in the supematants of different CTLs were detected by ELISA. Results： CEA610D peptide more efficiently induced CTL than CEA605-613 and MAGE-3 peptide did （P 〈 0.05）. The number of CD8＋ T cells in CEA610D group was significantly larger than those in CEA605-613 and MAGE-3 groups （P 〈 0. 05）. When the E： T was 40： 1, the cytotoxicity of CTLs induced by CEA610D and CEA605-613against CEA ＋ HLA-A2 ＋ T84 cells were （56.7 ± 3.73） % and （51.2 ± 1.86）%, respectively. But the CTL cytotoxicities induced by the three peptides against CEA ＋ HLA-A2-Lovo cells were all at the background level. Perforin expression and IFN-γ level of CTLs in CEA61oD group were significantly higher than those in the other two groups （P 〈 0.01 ）. Conclusion： Compared with natural CEA605-613 peptide, altered CEA60D peptide can effectively break the immune tolerance to self peptide, and thus has a stronger anti-tumor activity in vitro.

关 键 词：肿瘤抗原肽 癌胚抗原 修饰肽 细胞毒性T淋巴细胞 抗瘤作用 

分 类 号：R730.3[医药卫生—肿瘤] R730.51[医药卫生—临床医学]