树突状细胞疫苗对大鼠Walker-256肿瘤抑制作用.doc

树突状细胞疫苗对大鼠Walker-256肿瘤抑制作用
【摘要】目的建立Walker-256大鼠种植瘤模型,探讨树突状细胞(DC)疫苗抑制肿瘤生长的机制。方法从大鼠骨髓细胞中利用粒细胞-巨噬细胞集落刺激因子(GM-CSF)和白细胞介素4(IL-4)在体外定向诱导分化出DC,液氮冻融法提取Walker-256肿瘤细胞抗原后刺激DC制备负载抗原的DC疫苗。ELISA法检测培养液中IL-12的浓度。雌性SD大鼠,皮下注射Walker-256肿瘤细胞制备种植瘤模型,成瘤大鼠按照体重配对,分为肿瘤治疗组和肿瘤对照组。10只体重分别相对应的雌性SD大鼠作为正常对照组。肿瘤治疗组大鼠皮下注射负载抗原的DC细胞悬液,肿瘤对照组和正常对照组大鼠皮下注射等量PBS。分别检测大鼠血IL-12浓度和肿瘤最大径。结果得到大量具备典型光镜和电镜形态特征的DC,表达大鼠DC特异性标志OX62、OX6。负载抗原前后DC产生IL-12的水平存在明显差异(P<)。肿瘤治疗组大鼠肿瘤最大径显著小于肿瘤对照组(P<)。肿瘤对照组与正常对照组比较,IL-12明显下降(P<),肿瘤治疗组与肿瘤对照组比较,IL-12明显上升(P<)。肿瘤治疗组的肿瘤最大径与IL-12呈显著负相关(r=-,P<)。结论增强Th1介导的抗肿瘤细胞免疫可能是树突状细胞疫苗抑制肿瘤生长的机制之一。
【关键词】 Walker-256 树突状细胞肿瘤疫苗白细胞介素12
Inhibition of rat Walker-256 tumor growth by dendritic cell ine
WANG Jian1, YU Gang2, GUO Ji-qiang3, XU Gang2
( of General Surgery, the Second Affiliated Hospital of Xuzhou Medical College, Xuzhou,
Jiangsu 221006, China; 2004 of Postgraduate in General Surgery, Xuzhou Medical College,
Xuzhou, Jiangsu 221002; Clinical Laboratory, the Second Affiliated Hospital of Xuzhou Medical College)
Abstract: Objective To investigate the mechanism for dendritic cell (DC) ine to inhibit the growth of Walker-256 tumor in rats. Methods Soluble tumor antigen was prepared by running three freeze-thaw cycles of rat tumor cell line Walker-256. Anti-cancer DC ine was prepared by co-incubating the tumor antigen with the DC derived from rat bone marrow under the help of granulocyte/macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4). The level of IL-12 in culture medium was measred by ELISA. Female SD rats were injected Walker-256 tumor cells subcutaneously to form tumor models, which were paired on body weight and randomly divided into a treatment group and a tumor control group (n=10 each). Another 10 normal female SD rats were chosen to form the normal control group. DC ine(cell suspension of Walker-256 antigen sensitized DC)was injected subcutaneously to the treatment group, while an equal volume