摘要
目的:探讨伯氏疟原虫丝/苏氨酸磷酸酶5(PP5)作为传播阻断疫苗候选抗原的可行性。方法:PCR扩增PP5的功能区(407-711 aa),克隆入p ET32a(+)载体。IPTG诱导PP5重组蛋白表达后,纯化重组蛋白并免疫小鼠。ELISA方法检测抗PP5免疫血清效价。实验观察免疫血清对配子体出丝、动合子转化率和蚊胃内卵囊形成的影响。结果:成功表达PP5重组蛋白,抗PP5免疫血清抗体滴度可达1∶256 000,且对配子体出丝的抑制呈剂量依赖性,在1∶5和1∶10倍稀释时,配子体出丝数目分别减少75%和45%,与对照组相比差异具有显著统计学意义0. 05)。免疫血清可显著抑制动合子形成数目,在1∶5倍稀释时,动合子转化率减少14. 79%(P<0. 05),蚊感染率和胃内卵囊密度分别减少26. 05%和74. 19%(P<0. 05)。结论:PP5蛋白具有良好的免疫原性和抗原性。抗PP5免疫血清具有明显的传播阻断效果。
Objective:To investigate the possibility of selecting Plasmodium berghei serine/threonine phosphatase 5(PP5)as a transmission blocking vaccine candidate.Methods:PCR amplified the functional region of PP5(407-711 aa)and cloned into pET32a(+)vector.The recombinant PP5 protein was induced by IPTG,purified and immunized mice.ELISA detected the antibody titer of anti-PP5 serum.The effect of anti-PP5 sera on gametocyte exflagellation,ookinete conversion and oocyst formation in mosquito midgut was observed.Results:The recombinant PP5 protein was successfully expressed.The antibody titer of anti-PP5 serum was 1∶256 000,and the inhibition of gametocyte exflagellation was dose-dependent.The number of exflagellation significantly decreased by 75%and 45%at the anti-PP5 immune serum dilution of 1∶5 and 1∶10,respectively,compared with the control group(P<0.05).Meanwhile,the number of ookinete was significantly reduced on anti-PP5 sera treatment,and the ookinete conversion rate decreased by 14.79%(P<0.05),the prevalence of mosquito infection and oocyst density reduced 26.05%and 74.19%at 1∶5 times anti-PP5 sera dilution,respectively.Conclusion:PP5 protein has good immunogenicity and antigenicity.The anti-PP5 immune serum had obvious transmission blocking activity.
作者
孙林
洪明阳
曹雅明
朱晓彤
SUN Lin;HONG Ming-Yang;CAO Ya-Ming;ZHU Xiao-Tong(Department of Immunolgy,China Medical University,Shenyang 110122,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2019年第8期921-926,共6页
Chinese Journal of Immunology
基金
辽宁省教育厅课题(No.LQNK201727)资助
关键词
伯氏疟原虫
丝/苏氨酸磷酸酶5
传播阻断
配子体出丝
Plasmodium berghei
Serine/threonine protein phosphatase 5
Transmission blocking activity
Exflagellation
