摘要
目的:探讨约氏疟原虫蛋白磷酸酶6(PyPPM6)作为传播阻断疫苗候选抗原的可行性。方法:PCR扩增PyPPM6蛋白功能区(359-644 aa),克隆入pET32a(+)载体。IPTG诱导PyPPM6重组蛋白表达,免疫小鼠后收集抗-PyPPM6免疫血清。ELISA和Western Blot检测血清中抗体效价和特异性。体外实验观察免疫血清对配子体出丝、动合子形成和转化率及囊合子形成的影响。结果:成功表达PyPPM6重组蛋白;抗-PyPPM6免疫血清抗体滴度为1∶64000;与对照组相比,抗-PyPPM6免疫血清可使配子体出丝减少78.8%,动合子减少78.7%,动合子转化率降低55.1%,蚊胃内卵囊减少40.4%。结论:PyPPM6蛋白具有较强免疫原性。抗-PyPPM6免疫血清具有良好的传播阻断效果。
Objective:To determine the feasibility of selecting Plasmodium yoelii proteion phosphatase 6(PyPPM6)as a transmission blocking vaccine candidate.Methods:Functional domain(359-644 aa)of PyPPM6 was amplified by PCR and cloned into pET32a(+)vector.PyPPM6 recombinant protein(rPyPPM6)expression was induced by IPTG,anti-PyPPM6 serum was collected after immunizing mice.Serum titer and specificity of anti-PyPPM6 were detected by ELISA and Western blot.Effect of anti-PyPPM6 serum on exflagellation,ookinete formation and conversion,oocysts formation were investigated by in vivo experiment.Results:The recombinant PyPPM6 protein was successfully expressed,with an antibody titer around 1∶64000.Compared with control group,the exflagellation was significantly reduced by 78.8%on anti-PyPPM6 sera treatment.Ookinete formation decreased by 78.7%and conversion rate decreased by 55.1%in anti-PyPPM6 sera treatment group.And oocysts formation was significantly reduced by 40.4%in anti-PyPPM6 sera treatment group.Conclusion:PyPPM6 protein has good immunogenicity and antigenicity.The anti-PyPPM6 immune serum had obvious transmission blocking activity.
作者
杨慧琳
周丹
于园超
朱晓彤
崔立旺
YANG Hui-Lin;ZHOU Dan;YU Yuan-Chao;ZHU Xiao-Tong;CUI Li-Wang(Department of Immunology,China Medical University,Shenyang 110122,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2020年第12期1433-1436,1440,共5页
Chinese Journal of Immunology
基金
辽宁省教育厅课题(No.LQNK201727)资助。
关键词
约氏疟原虫
蛋白磷酸酶6
传播阻断
有性发育
Plasmodium yoelii
Protein phosphatase 6
Transmission blocking
Sexual stage development
