猪带绦虫14-3-3.3重组蛋白疫苗诱导小鼠的免疫应答  

Immune response induced by 14-3-3.3 recombinant protein vaccine of Taenia solium in mice

作　　者：严海东 邹旭东 张一雯 周必英 Yan Haidong;Zou Xudong;Zhang Yiwen;Zhou Biying(Department of Parasitology,Zunyi Medical University,Zunyi 563000,China)

机构地区：[1]遵义医科大学寄生虫学教研室,563000

出　　处：《中华地方病学杂志》2021年第10期794-801,共8页Chinese Journal of Endemiology

基　　金：贵州省科技计划项目(黔科合基础[2018]1190);遵义医科大学大学生创新创业训练计划项目(ZYDC2018056)。

摘　　要：目的探究猪带绦虫(Ts)14-3-3.3蛋白作为囊虫病疫苗候选分子的潜力。方法选取60只昆明小鼠,体重为18~22 g,按体重采用随机数字表法分为3组,分别为生理盐水对照组(对照组)、Ts14-3-3.3重组蛋白疫苗组(疫苗组)、Ts14-3-3.3重组蛋白疫苗+佐剂组(疫苗+佐剂组),每组20只。采用多点皮下注射法,在0周首次免疫后进行2次加强免疫,共免疫3次,每次接种间隔2周。3组在首次免疫后0、2、4、6、8周分别剖杀4只小鼠,摘眼球取血和无菌取脾,分别用于分离血清和制备脾淋巴细胞悬液[处理因素:原液、抗原刺激、刀豆蛋白A(ConA)刺激]。采用间接酶联免疫吸附试验(ELISA)法检测小鼠血清特异性抗体免疫球蛋白(Ig)G、IgG2a、IgG1及IgE水平;采用CCK-8法检测小鼠脾淋巴细胞增殖水平;采用双抗夹心ELISA法检测小鼠脾淋巴细胞培养上清液肿瘤坏死因子(TNF)-α、白细胞介素(IL)-12、IL-13、IL-10水平。结果疫苗、疫苗+佐剂组免疫2~8周IgG、IgG2a、IgG1水平均高于对照组,且疫苗+佐剂组免疫2~8周上述指标均高于疫苗组(P均<0.05)。组间相同处理因素下,免疫2~8周脾淋巴细胞增殖水平以及培养上清液TNF-α、IL-12、IL-13、IL-10水平比较,差异均有统计学意义(P均<0.05);疫苗、疫苗+佐剂组免疫2~8周脾淋巴细胞增殖水平以及培养上清液TNF-α、IL-12、IL-13、IL-10水平均高于对照组,且疫苗+佐剂组免疫2~8周上述指标均高于疫苗组(P均<0.05)。组内不同处理因素下,抗原刺激、ConA刺激时免疫0~8周脾淋巴细胞增殖水平以及培养上清液TNF-α、IL-12、IL-13、IL-10水平均高于原液,且ConA刺激时免疫0~8周上述指标均高于抗原刺激(P均<0.05)。结论Ts14-3-3.3重组蛋白疫苗可诱导小鼠产生有效的免疫应答。Objective To explore the potential of Taenia solium(Ts)14-3-3.3 protein as a candidate molecule for cysticercosis vaccine.Methods Sixty Kunming mice with the body weight of 18-22 g were selected and divided into 3 groups according to their body weight via the random number table method,including normal saline control group(control group),Ts14-3-3.3 recombinant protein vaccine group(vaccine group),and Ts14-3-3.3 recombinant protein vaccine+adjuvant group(vaccine+adjuvant group),with 20 mice in each group.The multi-point subcutaneous injection method was adopted.After the first immunization at 0 week,the booster immunization was carried out twice,a total of 3 times,with an interval of 2 weeks.Four mice in the three groups were killed at 0,2,4,6 and 8 weeks after the first immunization,and the blood of eyeballs and spleen were collected aseptically for serum separation and preparation of spleen lymphocytes suspension[treatment:cell suspension,antigen-stimulate and concanavalin(Con)A-stimulate],respectively.The levels of mouse serum specific immunoglobulin(Ig)G,IgG2a,IgG1 and IgE were detected by indirect enzyme-linked immunosorbent assay(ELISA).The proliferation level of mouse spleen lymphocytes was detected via the CCK-8 method.The levels of tumor necrosis factor(TNF)-α,interleukin(IL)-12,IL-13 and IL-10 in culture supernatant of mouse spleen lymphocytes were determined by double-antibody sandwich ELISA.Results The IgG,IgG2a,and IgG1 levels of the vaccine and vaccine+adjuvant groups immunized for 2 to 8 weeks were higher than those of the control group,and the above indicators of the vaccine+adjuvant group were higher than those of the vaccine group(P<0.05).With the same treatment between the groups,the proliferation levels of spleen lymphocytes and the levels of TNF-α,IL-12,IL-13,and IL-10 in the culture supernatant after 2-8 weeks of immunization were statistically significantly different(P<0.05);the proliferation levels of spleen lymphocytes and the levels of TNF-α,IL-12,IL-13 and IL-10 in the culture superna

关 键 词：猪带绦虫 猪囊尾蚴 Ts14-3-3.3重组蛋白疫苗 免疫应答 

分 类 号：R392[医药卫生—免疫学]