弓形虫MIC1、MIC6和ROP18混合蛋白对小鼠的免疫保护作用  

Immunological Protection of MIC1-MIC6-Rop18 Recombinant Vaccine Against Toxoplasma gondii in Mice

作　　者：李虎 朱进进 李婧旸 韩成建 余莉[1] LI Hu;ZHU Jinjin;LI Jingyang;HAN Chengjian;YU Li(Key Laboratory of Microbiology and Parasitology of Anhui Province,Key Laboratory of Zoonoses of High Institutions in Anhui,Department of Microbiology and Parasitology,School of Basic Medical Sciences,Anhui Medical University,Hefei 230032,China;Clinical Laboratory,The Third People’s Hospital of Hefei,Hefei 230032,China)

机构地区：[1]病原生物学安徽省重点实验室,安徽省高校人畜共患病重点实验室,安徽医科大学基础医学院微生物学与寄生虫学教研室,合肥230032 [2]安徽医科大学合肥第三临床学院合肥市第三人民医院检验科,合肥230032

出　　处：《中国动物传染病学报》2024年第3期34-41,共8页Chinese Journal of Animal Infectious Diseases

基　　金：国家自然科学基金(81871671,82072304)。

摘　　要：为评估弓形虫微线体蛋白MIC1、MIC6和棒状体蛋白ROP18组成的重组疫苗对BALB/c小鼠的免疫保护作用,我们将重组表达的MIC1、MIC6和ROP18三种蛋白进行不同的组合,并联合弗氏佐剂通过皮下注射的方式免疫小鼠。用ELISA技术检测免疫后小鼠血清特异性抗体,以及免疫蛋白刺激后脾细胞培养上清中各细胞因子的水平,通过脾淋巴细胞增殖实验检测脾淋巴细胞增殖情况。最后一次免疫后,用弓形虫WH3强毒株攻击,观察并记录小鼠的存活情况。结果表明,与对照PBS组相比,蛋白免疫组产生高滴度的IgG和IgG亚型(IgG1和IgG2a)抗体,滴度可高达1∶128000。细胞因子IL-4、IL-10和IFN-γ表达水平不同程度地升高,并且重组蛋白能够诱导淋巴细胞特异性增殖。免疫组小鼠的生存时间较对照组有所延长,其中MIC6-ROP18组延长时间最长(P<0.01)。MIC1、MIC6和ROP18组成的重组疫苗能诱导小鼠产生较强的体液免疫和细胞免疫应答,其中MIC6-ROP18蛋白组合的免疫保护效果更为明显,但仍需进一步改进。本研究为弓形虫疫苗的研发提供了思路。To evaluate the immune protection of recombinant protein vaccine composed of Toxoplasma gondii microneme proteins 1,6(MIC1,MIC6)and rhoptry protein 18(ROP18)in BALB/c mice,different combinations of those proteins were emulsified with Freund’s adjuvant and used to subcutaneously immunize mice.The specific antibodies in mouse serum samples and cytokines levels in the cultural supernatant of splenocytes were tested using ELISA method.After the final immunization,mice were challenged with virulent T.gondii WH3 strain and the survival time was recorded.The results showed that the mice immunized with the recombinant proteins produced higher titers of IgG,IgG1 and IgG2a than those in PBS control group and the titer reached above 1:128000.The levels of IL-4,IL-10 and IFN-γin the immune groups increased to varying degrees as compared with PBS control group.Meanwhile,these recombinant proteins induced specific lymphocyte proliferation.Moreover,the survival time of immunized mice was prolonged than control group.Taken together,the results in the present study indicated that vaccination with the recombinant MIC1,MIC6 and ROP18 proteins elicited strong humoral and cellular immune responses in mice,in which MIC6-ROP18 protein combination had a more obvious immune protection effect but needed further investigation.Our study provided a reference for the development of T.gondii vaccine.

关 键 词：弓形虫疫苗 重组蛋白 MIC1 MIC6 ROP18 

分 类 号：S852.4[农业科学—基础兽医学]