Jiao Jie-Ge, Zhang Yang-De, Li Yue-Nan
Hepatobiliary and Enteric Surgical Research Center of Health Ministry, Center South University, Changsha, Hunan, P. R. China.
Ai Zheng. 2005 Oct;24(10):1179-83.
BACKGROUND & OBJECTIVE: Endoglin is a marker of tumor angiogenesis. Increasing evidences proved that passive immunotherapy with anti-endoglin monoclonal antibody can effectively inhibit tumor growth, and xenogeneic homologous DNA vaccine can inhibit cross antitumor immunity. This study was to explore the inhibitory effect of a DNA vaccine encoding the extracellular domain of porcine endoglin (ppEDG) on tumor growth in a mouse colon carcinoma model.
ppEDG was used as a DNA vaccine to actively immunize the colon carcinoma-bearing mice. Tumor volume and survival rate of the mice were observed in 3-day intervals. Microvessel density (MVD) was detected by immunohistochemistry; antibodies against self-endoglin were detected by Western blot and ELISA; the B cells that secrete auto-antibodies against self-endoglin were detected by ELISPOT assay.
Eighteen days after tumor cell inoculation, the tumor volume was significantly smaller in ppEDG-immunized group than in empty plasmid (e-p) group and normal saline (NS) group (P<0.05), and the survival time was significantly longer in ppEDG-immunized group than in the control groups (P<0.001). MVD was significantly lower in ppEDG-immunized group than in e-p group and NS group (19.2+/-4.5 vs. 76.9+/-14.4 and 81.4+/-16.9, P<0.001). The antibodies against self-endoglin were identified in ppEDG-immunized group; the major subtypes were IgG(1) and IgG(2b). The auto-antibody-producing B cells were much more in ppEDG-immunized group than in e-p group and NS group (82.5+/-14.1 vs. 3.6+/-1.3 and 4.7+/-2.0, P<0.001).
ppEDG DNA vaccine could induce the production of auto-antibodies against self-endoglin, which further inhibit angiogenesis and growth of colon carcinoma.
内皮糖蛋白是肿瘤血管生成的标志物。越来越多的证据表明,抗内皮糖蛋白单克隆抗体被动免疫疗法可有效抑制肿瘤生长,而异基因同源DNA疫苗可抑制交叉抗肿瘤免疫。本研究旨在探讨编码猪内皮糖蛋白胞外域(ppEDG)的DNA疫苗对小鼠结肠癌模型肿瘤生长的抑制作用。
用ppEDG作为DNA疫苗主动免疫荷结肠癌小鼠。每隔3天观察小鼠的肿瘤体积和存活率。采用免疫组化法检测微血管密度(MVD);采用蛋白质印迹法和酶联免疫吸附测定法检测抗自身内皮糖蛋白抗体;采用酶联免疫斑点分析法检测分泌抗自身内皮糖蛋白自身抗体的B细胞。
接种肿瘤细胞18天后,ppEDG免疫组的肿瘤体积明显小于空质粒(e-p)组和生理盐水(NS)组(P<0.05),存活时间明显长于对照组(P<0.001)。ppEDG免疫组的MVD明显低于e-p组和NS组(19.2±4.5对76.9±14.4和81.4±16.9,P<0.001)。在ppEDG免疫组中鉴定出抗自身内皮糖蛋白抗体;主要亚型为IgG(1)和IgG(2b)。ppEDG免疫组中产生自身抗体的B细胞比e-p组和NS组多得多(82.5±14.1对3.6±1.3和4.7±2.0,P<0.001)。
ppEDG DNA疫苗可诱导产生抗自身内皮糖蛋白的自身抗体,进而抑制结肠癌的血管生成和生长。