细粒棘球绦虫重组Bb-Eg95-EgA31疫苗诱导小鼠脾细胞因子变化的研究

目的探讨细粒棘球绦虫(Eg)重组双歧杆菌(Bb)-Eg95-EgA31疫苗免疫和Eg原头节攻击后小鼠脾细胞因子的变化。方法 56只雌性BALB/c小鼠随机均分7组,A组和B组分别皮下和肌肉注射5×106蛋白克隆形成单位(CFU)疫苗悬浮于100μl双歧杆菌液体培养基(MRS)、C组鼻腔内接种5×105CFU+10μlMRS、D组经口灌胃5×108CFU+100μlMRS、E组(空载体对照组)5×106CFUBb(pGEX-1λT)+100μlMRS、F组为双歧杆菌(Bb)对照组和G组为双歧杆菌液体培养基(MRS)对照组。免疫后8周各组鼠攻击感染50个Eg原头节,感染后25周剖杀小鼠,取脾,分离脾细胞,用Eg粗抗原(EgAg)刺激培养,同时设伴刀豆球蛋白A(ConA)或脂多糖(LPS)刺激对照和非刺激组。收集脾细胞培养上清液,ELISA法检测脾细胞培养上清液中γ干扰素(IFN-γ)、白细胞介素-12(IL-12)、α肿瘤坏死因子(TNF-α)和IL-10水平。结果 A、B、C和D组小鼠脾细胞培养上清液中,IFN-γ水平分别为(137.5±23.2)、(162.5±23.2)、(250.0±53.5)和(215.0±37.4)pg/ml,均高于MRS对照组(50.0±10.7)pg/ml(P0.01);IL-12水平分别为(27.5±4.6)、(32.5±4.6)、(45.0±5.4和(35.0±5.4)pg/ml,均高于MRS对照组(15.0±5.4)pg/ml(P0.01);TNF-α水平分别为(275.0±46.3)、(325.0±46.3)、(450.0±53.5)和(350.0±53.5)pg/ml,均高于MRS对照组(150.0±53.5)pg/ml(P0.01);IL-10水平分别为(37.5±4.6)、(35.0±5.4)、(25.0±5.4)和(32.5±4.6)pg/ml,均低于MRS对照组(45.0±5.4)pg/ml(P0.05或P0.01)。结论细粒棘球绦虫重组Bb-Eg95-EgA31疫苗可诱导小鼠产生Th1型保护性免疫应答。

Changes of spleen cytokines in mice immunized with recombinant Bb-Eg95-EgA31 vaccine against Echinococcus granulosus

Objective To investigate the level of cytokines in spleen from mice immunized with recombinant Bifidobacteria bifidum（Bb）-Eg95-EgA31 vaccine of Echinococcus granulosus（Eg） and challenged with Eg protoscoleces. Methods 56 female BALB/c mice were randomly divided into 7 groups: Recombinant Bb-Eg95-EgA31 vaccine subcutaneous injection （group A）, intramuscular injection （B）, intranasal immunization （C）, oral administration （D）, blank vector control （E）, Bb control （F） and MRS control （G）. Mice in all groups were challenged with 50 Eg protoscoleces on the 8th week after vaccination and sacrificed on the 25th week after infection. Spleens were collected to prepare splenocytes, which were cultured under activation of crude Eg antigen （EgAg）, with concanavalin A （ConA） or lipopolysaccharide （LPS） stimulation and non-stimulation as control. The splenocyte supernatants were collected to determine the levels of interferon-γ（IFN-γ）, interleukin-12 （IL-12）, tumour necrosis factor-α （TNF-α） and IL-10 by ELISA. Results The level of IFN-γ in groups A ［（137.5±23.2） pg/ml］, B ［（162.5±23.2） pg/ml］, C ［（250.0±53.5） pg/ml］ and D ［（215.0±37.4） pg/ml］ was higher than that of group G ［（50.0±10.7） pg/ml］ （P＜0.01）. The level of IL-12 in groups A ［（27.5±4.6） pg/ml］, B ［（32.5±4.6） pg/ml］, C ［（45.0±5.4） pg/ml］ and D ［（35.0±5.4） pg/ml］ was higher than that of group G ［（15.0±5.4） pg/ml］ （P＜0.01）. The level of TNF-α in groups A ［（275.0±46.3） pg/ml］, B ［（325.0±46.3） pg/ml］, C ［（450.0±53.5） pg/ml］ and D ［（350.0±53.5） pg/ml］ was higher than that of group G ［（150.0±53.5） pg/ml］ （P＜0.01）. The level of IL-10 in groups A ［（37.5±4.6） pg/ml］, B ［（35.0±5.4） pg/ml］, C ［（25.0±5.4） pg/ml］ and D ［（32.5±4.6） pg/ml］ was lower than that of group G ［（45.0±5.4） pg/ml］ （P＜0.05 or P＜0.01）. Conclusion The recombinant Bb-Eg95-EgA31 vaccine can induce a protective Th1 type immune response in mice.