【摘要】 目的构建靶向树突状细胞(dendritic cell,DC)的肿瘤疫苗DC-OVA系统,探讨DC-OVA肿瘤疫苗在祼鼠体内的抗肿瘤效应及相关机制。方法构建表达肿瘤抗原OVA的树突状细胞。建立携带OVA的Lewis肺癌细胞(Lewis lung cancer cell-OVA,LLC-OVA)的皮下荷瘤裸鼠模型,瘤旁注射DC-OVA疫苗,通过记录肿瘤大小观察肿瘤生长抑制状况。以IHC方法观察肿瘤组织CD3、Nestin、CD133阳性细胞;观察脾DC相关分子(CD11c、OVA)的阳性表达以及T细胞(CD3+)的分布、数量和功能变化。同时,应用ELISA方法检测血清中IL-12的表达水平。结果 DC-OVA疫苗预防组与PBS对照组比较,呈现出明显的肿瘤生长抑制(P<0.05)。与对照组相比,DC-OVA疫苗治疗组肿瘤明显缩小(P<0.001)。疫苗治疗3次组与治疗1次组比较,效果更明显(P<0.05)。于实验第4周,疫苗治疗1次组与对照组比较,CD3阳性表达高,Nestin、CD133阳性表达低(P<0.05);治疗1次组血清IL-12含量较对照组升高(P<0.001);第8周,治疗1次组血清IL-12含量较第4周治疗组及第8周对照组低。结论 DC-OVA疫苗特异性杀伤了LLC-OVA肿瘤细胞,使肿瘤干细胞表达Nestin、CD133减少。更多还原

【Abstract】 Objective To prepare DC-OVA vaccine system and to investigate its anti-tumor effect in Lewis lung cancer bearing nude mice. Methods Ovalalbumin( OVA) plasmid was transfected into dendritic cells( DC) to construct DC-OVA and cytotoxic T cells were activated by DC-OVA. The DC-OVA vaccine system was prepared. Lewis lung cancer cells + OVA were inoculated in nude mice,and the tumor-bearing( LLC-OVA) nude mice were treated with DC-OVA. The tumor growth was measured by vernier caliper; the expression of CD11 c,OVA and CD3 in mice spleen,the expression of CD3,Nestin and CD133 in tumor tissue were detected by immunohistochemistry( IHC); serum IL-12 in nude mice was measured by ELISA. Results Compared with control group,vaccine prevention group presented significant inhibition effect on tumor growth( P < 0. 05). Compared with control group the tumor size was significantly smaller in vaccine treatment group( P < 0. 001). The tumor-inhibition effect in nude mice with 3-time treatment was more marked than that with one-time treatment( P <0. 05). In week 4 compared with control group,the expression of CD3 in vaccine treatment group was higher and expression of Nestin and CD133 was lower( P < 0. 05). The serum IL-12 in vaccine treatment group was significantly increased( P < 0. 001) at week 4; however,serum IL-12 in treatment group at week 8 was lower than that in treatment group at week 4 and control group at week 8( both P < 0. 001). Conclusion DC-OVA vaccine system can prevent and treat tumor-bearing nude mice,and it is effective in inhibiting tumor stem cells expressing Nestin and CD133.更多还原