MUC1肿瘤抗原表位PDTRP基因免疫诱导的体内外抗瘤效应

Anti-Tumor Efficacy Induced by Antibodized PDTRP Gene Immunization

目的:构建MUC1肿瘤抗原表位γlneo-PDTRP质粒,基因免疫诱导机体产生有效的体内外抗瘤效应。方法:用 Insight Ⅱ将PDTRP抗原表位与MUC1分子模拟,构建含PDTRP的抗原化抗体表达质粒γlneo-PDTRP,体外转染检测表达。 用该质粒基因免疫小鼠,免疫后ELISA动态检测血清中抗PDTRP抗体的产生及特异性细胞免疫应答;以4T1-PDTRP肿瘤细 胞攻击经γlneo-PDTRP免疫小鼠,观察免疫保护效应。结果:三串联肿瘤抗原表位PDTRPDTRPDTRP模拟了MUC1核心序列 的空间结构;构建γlneo-PDTRP质粒并在体外转染表达,基因免疫小鼠诱导出特异性抗PDTRP体液和细胞免疫应答,免疫小 鼠对4T1-PDTRP肿瘤细胞的攻击具有显著的体内保护效应。结论:MUC1肿瘤抗原表位PDTRP可模拟MUC1核心序列的天 然构象,γlneo-PDTRP基因免疫在体外诱导特异性的体液和细胞免疫应答,体内诱导对机体的免疫保护效应。

Objective: To study the anti-tumor efficacy induced by antibodized tumor epitope PDTRP gene immunization. Methods: Three copies of tumor associate gene PDTRP from MUCI tandem repeats were designed and mimicked the conformation of MUCI by Insight Ⅱ . The γlneo-PDTRP plasmid was further constructed, in which the PDTRP target gene was inserted into CDR3 of the γ1 -neo vector. The specific humoral and cellular immune responses towards to PDTRP were detected after intraspleen immunized Balb/c mice with 'ylneo-PDTRP. And the immune protection assay was also done to observe whether the mice immunized with γlneo-PDTRP could prolong the survival after tumor challenge. Results: The conformation of three copies of PDTRP mimicked the conformation of MUCI tandem repeats. The expression of γlneo-PDTRP could be detected after in vitro transfect. The specific antibody against PDTRP epitope could be induced and increase to a higher titer after intraspleen injection with a γlneo-PDTRP plasmid. And the specific proliferation and cytotox-ic function of lymphocyte were also increased. There is a significant survival from mice immunized with γlneo-PDTRP a-gainst the 4T1-PDTRP tumor challenge. Conclusions: Gene immunization with γlneo-PDTRP could elicit both humoral and cellular tumor specific immune response and had the protective effect.