磷酸钙纳米颗粒修饰的甲胎蛋白和树突状细胞疫苗对小鼠皮下移植瘤的抑制作用  

Inhibition of the growth of hepatocarcinoma xenografl in Balb/c mice induced by dendritic cells immunized with AFP cDNA fragement

作　　者：曾斌[1] 廖爱军[1] 卢放根[2] 方唯意 王健 

机构地区：[1]南华大学附属第一医院消化内科,衡阳421001 [2]中南大学湘雅二医院消化内科 [3]湘雅医学院肿瘤研究所

出　　处：《中华肿瘤杂志》2010年第2期98-102,共5页Chinese Journal of Oncology

基　　金：国家自然科学基金（39570335）;湖南省自然科学基金（08JJ3032）

摘　　要：目的探讨转染小鼠带有信号肽的AFP1cDNA和去掉信号肽的AFP2cDNA树突状细胞（DC）在体外的免疫活性，以及其对Balb／e小鼠皮下移植瘤的抑制作用。方法应用磷酸钙纳米颗粒将带有信号肽的AFP1cDNA和去掉信号肽的AFP1cDNA真核表达载体pcDNA3．1转染DC，将DC疫苗与同源小鼠脾淋巴细胞混合培养，采用酶联免疫吸附（ELISA）法，检测上清液中干扰素-γ（IFN-γ）的表达情况。采用四甲基偶氮唑蓝（MTT）法，检测AFP1／DC和AFP2／DC刺激同基因小鼠脾淋巴细胞增殖能力及诱导特异性细胞毒性T淋巴细胞（CTL）的杀伤能力。观察AFP1／DC和AFP2／DC对Balb／e小鼠皮下移植瘤生长的抑制作用。结果AFP2／DC能明显促进脾淋巴细胞增殖并提高CTL的特异性杀伤作用。AFP1／DC和AFP2／DC瘤内注射均可显著抑制肝癌移植瘤的生长，但AFP2／DC的抑制作用更明显，治疗2周后，AFR／DC组小鼠肿瘤体积为（726．7±298．2）mm3，明显小于AFP2／DC组[（1486．2±457．2）mm3]和空质粒对照组[（2137．2±547．2）mm3，P〈0．05]。AFR／DC组和AFP，／DC组的抑瘤率分别达79．2％和39．7％，而空质粒对照组和空白对照组的抑瘤率为0。AFP2／DC组和AFP，／DC组小鼠的生存时间分别为（58．5±4．2）d和（45．2±4．8）d，较空质粒对照组[（30．6±6．2）d]显著延长（P〈0．05）。结论AFP2／DC疫苗在体外能够诱导出高效而特异的抗肿瘤免疫效应，在体内具有抑制Balb／c小鼠皮下移植瘤生长的作用。Objective To investigate the antitumor immune response induced by dendritic cells vaccine coding AFPcDNA fragment with signal peptide （ AFP1 ） and without signal peptide （ AFP2 ）, and to determine the inhibiting effect of the vaccine on the growth of hepatocarcinoma xenograft in Balb/c mice. Methods pcDNA3.1/AFP~ and pcDNA3.1/AFP2 were transfected into dendritic cells （DCs） by calcium phosphate nanoparticles and became DCs vaccine. Mouse spleen lymphocytes were stimulated by AFPJDC and AFPJI）C. A Balb/c mouse model bearing mouse HCC xenograft was established on the day 14 after transplantation. Forty mice were divided equally into AFP2/DC group, AFP1/DC group and plasmid control group. The treated mice received DCs vaccine and the same amount of control plasmid. Results AFP2/DC stimulated T lymphocytel proliferation in vitro and improved CTL activity. The effects were better than AFP1/DC. The tumor-bearing mice injected intralesionally with AFP1/DC and AFP2/DC at a dose of 0. 5 rnl per mouse showed inhibition of tumor growth and prolongation of survival time. The tumor inhibition rate of the AFP2/DC group was 79.2% and the AFP1/DC group was 39.7% at 2 weeks after treatment. The tumor volume of AFPJDC group was （726.7 ±298.2 ）mm3, significantly smaller than the （ 1486.2± 457.2 ） mm3 of the AFP1/DC group and （2137.2 ± 547.2） mm3 of the plasmid control group （P 〈 0.05 ）. The mean survival time of mice in the AFPJDC group [（58.5±4.2）d1 and AFP1/DC group [（45. 2 ±4. 8） d] were significantly longer than that of plasmid control group [ （ 30. 6 -＋ 6. 2 ） d, P 〈 0. 05 ]. Bax-positive cell percentage was increased in the xenografts of AFP2/DC-treatment group compare with that of plasmid control group. Conclusion AFP2/DC and AFP~/DC vaccines show evident inhibiting effect on the growth of H22 xenograft in Balb/c mice through inducing efficient and specific immune response against the hepatocarcinoma ceils.

关 键 词：甲胎蛋白 肝肿瘤 疫苗 树突状细胞 小鼠 

分 类 号：R735.7[医药卫生—肿瘤]