多房棘球绦虫重组BCG-EmⅡ/3疫苗诱导小鼠脾细胞因子变化的研究

目的 探讨多房棘球绦虫(Em)重组BCG-EmⅡ/3疫苗免疫和Em原头节攻击后小鼠脾细胞因子的变化.方法 将Balb/c小鼠按体质量随机分为3组:疫苗皮下注射组、疫苗鼻腔内接种组、对照组.免疫后8周用Em原头节进行攻击感染,感染后18周杀鼠取脾,分离脾细胞,分别用原液、Em抗原(EmAg)、刀豆蛋白A(ConA)或植物血凝素(PHA)培养,酶联免疫吸附(ELISA)法检测脾细胞培养上清液中白细胞介素2(IL-2)、γ干扰素(IFN-γ)、肿瘤坏死因子α(TNF-β)、白细胞介素4(IL-4)水平.结果 疫苗皮下注射组原液培养条件下IL-2、IFN-γ、TNF-α和IL-4水平分别为(34.6±2.7)、(34.5±2.8)、(265.0±0.0)、(9.8±2.6)ng/L,与对照组[(25.0±1.9)、(30.0±0.0)、(10.0±0.0)、(12.5±2.7)ng/L]比较差异均有统计学意义(P＜0.01或＜0.05);疫苗鼻腔内接种组原液培养条件下上述4种指标分别为(32.5±2.2)、(33.6±2.7)、(130.0±0.0)、(10.4±2.7)ng/L.与对照组比较差异均有统计学意义(P＜0.01或＜0.05);疫苗皮下注射组原液培养条件下TNF-α水平明显高于疫苗鼻腔内接种组(P＜0.01).EmAg、ConA(或PHA)刺激培养条件下,上述4种指标明显高于相同组别的原液培养(P＜0.01),且ConA(或PHA)刺激培养条件下,上述4种指标明显高于相同组别的EmAg刺激培养条件(P＜0.01).结论 多房棘球绦虫重组BCG-EmⅡ/3疫苗诱导小鼠产生辅助T淋巴细胞(Th)1型免疫反应,从而对抗Em原头节攻击感染.

Study on the changes of cytokines of splenocytes in mice by immunization with recombinant BCG-Em Ⅱ/3 vaccine against Echinococcus multilocularis

Objective To investigate the changes of cytokines of splenocytes in mice immunized with recombinant BCG-Em Ⅱ/3 vaccine of Echinococcus multilocularis(Em)and consequently challenged with Em protoscoleces.Methods Balb/c mice were subcutaneously or intranasally vaccinated and challenged with Em were separated and cultured with EmAg,ConA or PHA,respectively.The supematants were gathered to measure the levels of IL-2,IFN-γ,TNF-α.and IL-4 by ELISA Kits.Results The levels of IL-2,IFN-γ,TNF-α and IL-4 in the subcutaneous group were(34.6±2.7),(34.5±2.8),(265.0 ±0.0)and(9.8±2.6)ng/L respeetively:those in the intranasal group were(32.5±2.2),(33.6±2.7),(130.0±0.0)and(10.4±27)ng/L respectively;those in the control were(25.0±1.9),(30.0±0.0),(10.0±0.0)and(12.5±2.7)ng/L,respectively:there were statistical differences between the immunized groups and control group(P＜0.01 or＜0.05);The level of TNF-α in the subcutaneous group was higher than that in the intranasal group.Conclusion Th1 response has been induced in mice vaccinated with rBCG-Em Ⅱ/3 vaccine and challenged with Em protoscoleces.