中文摘要：

目的动态观察细粒棘球绦虫（喀）转Eg95-EgA31融合基因苜蓿疫苗免疫Balb／c小鼠后脾细胞因子的变化。方法88只Balb／c小鼠按体质量随机分为两组，用转基因苜蓿叶蛋白提取液（20g／L）100μl口服灌胃和10μl鼻腔黏膜接种分别免疫小鼠，每3天1次，连续2个月。在末次免疫后0（对照）、2、4、6、8、10、12、14、16、18、20周各组剖杀4只小鼠，取脾，分离脾细胞，体外经瞻粗抗原（EgAg）或刀豆蛋白A（ConA）刺激培养48h，诱生白细胞介素（IL）-12、γ-干扰素（IFN-γ）和IL-10；经EgAg或脂多糖（LPS）刺激培养72h，诱生肿瘤坏死因子仅（TNF-α）。收集脾细胞培养上清液，常规ELISA法检测IL-12、IFN-γ、TNF-α和IL-10水平。结果口服灌胃组的IL-12、IFN-γ、TNF—α．和IL-10水平分别在免疫后4～6、2～8、2～6、4～12周升高，分别在免疫后4、2、2、8周达最高水平，其值分别为（25．0±5．8）、（575．0±28．9）、（50．0±11．5）、（42．5±2．9）ng／L，与0周[（11．3±2．5）、（125．0±28．9）、（11．3±2．5）、（12．5±2．9）ng／L]比较，差异有统计学意义（P均〈0．01）；鼻腔黏膜接种组的IL-12、IFN-γ、TNF-α和IL-10水平分别在免疫后4—6、2～10、4～10、6—16周升高，分别在免疫后6、4、6、6周达最高水平，其值分别为（25．0±5．8）、（725．0±28．9）、（27．5±2．9）、（60．0±11．5）ng／L，与0周[（11．3±2．5）、（125．0±28．9）、（11．3±2．5）、（12．5±2．9）ng／L]比较，差异有统计学意义（P均〈0．01）。EgAg、ConA、LPS刺激组的细胞因子水平高于相应的脾细胞悬液组（P〈0．05或〈0．01），ConA或LPS刺激组的细胞因子水平高于相应的EgAg刺激组（P〈0．05或〈0．01）。结论细粒棘球绦虫转Eg95-EgA31融合基因苜蓿疫苗在免疫早期（2～10周）可诱导小鼠产生Th1和Th2混合型免疫应答。

英文摘要：

Objective To investigate the dynamic changes of splenoeyte cytokines in mice immunized with the transgenic alfalfa containing Eg95-EgA31 fusion gene of Eehinococcus granulosus （Eg）. Methods Eighty-eight Balb/e mice were divided into 2 groups randomly according to body weights, and immunized orally or intranasally with 100 μl or 10μl extracted leaf protein from the transgenic affalfa（20 g/L） respectively once per 3 days for 2 months. Four mice randomized from each group were killed to get splenocyte on week O（control）,2,4,6,8, 10,12,14,16,18 and 20 after the last immunization. The splenocyte were cultured in medium for 48 hours with EgAg or eoncanavalin A （ConA） stimulation to induce the interleukin （ IL ）- 12, interferon γ（IFN-γ） and IL- 10, and cultured for 72 hours with EgAg or lipopolysaceharide（LPS） stimulus to induce the tumor necrosis factor α （TNF-α）. Then the supernatant was collected to measure the level of IL-12, IFN-γ, TNF-α and IL-10 by ELISA. Results In the oral immunization group, the level of IL-12, IFN-γ, TNF-α and IL-10 increased significantly from week 4 to week 6, week 2 to week 8, week 2 to week 6 and week 4 to week 12, respectively, reaching the highest level（25.0 ± 5.8）ng/L on week 4, （575.0 ± 28.9）ng/L on week 2, （50.0 ± 11.5）ng/L on week 2 and（42.5 ± 2.9）ng/L on week 8, respectively, as compared with the values on week 0 [ （11.3± 2.5）, （125.0 ± 28.9）, （11.3 ± 2.5）, （12.5 ± 2.9）ng/L, all P〈 0.011 ; in the intranasal immunization group, it was similar about the values of IL-12, IFN-γ, TNF-α and IL-10 could be seen from week 4 to week 6, week 2 to week 10, week 4 to week 10 and week 6 to week 16, respectively, reaching the highest level（25.0 ± 5.8）ng/L on week 6, （725.0 ± 28.9）ng/L on week 4, （27.5 ± 2.9）ng/L on week 6 and （60.0 ± 11.5）ng/L on week 6, respectively, as compared with the values on week 0[ （ 11.3 ± 2.5 ）, （ 125.0 ± 28.9 ）, （ 11.3 ± 2.5 ）, （ 12.5 ± 2.9）ng/