约氏疟原虫丝/苏氨酸磷酸酶5抗血清对有性阶段生长抑制作用的研究

Inhibitive effect of Plasmodium yoelii serine/threonine phosphatase 5 on sexual stage development

目的:探讨约氏疟原虫丝/苏氨酸磷酸酶5(PyPP5)作为传播阻断疫苗候选抗原的可行性。方法:PCR扩增PyPP5蛋白优势抗原表位,克隆入pET32a(+)载体。IPTG诱导PyPP5重组蛋白(rPyPP5)表达后,纯化并免疫小鼠获取抗血清。体外实验观察抗-PyPP5免疫血清对配子体出丝、动合子形成和转化率的影响。结果:成功制备抗-PyPP5免疫血清。抗-PyPP5免疫血清(1∶5倍稀释)使配子体出丝率、动合子数目和动合子转化率分别下降29.89%(P<0.05)、38.59%(P<0.05)和64.30%(P<0.01)。结论:PyPP5蛋白具有良好的抗原性。抗-PyPP5免疫血清可有效抑制约氏疟原虫传播。

Objective:To investigate the possibility of selecting Plasmodium yoelii serine/threonine phosphatase 5(PyPP5)as a transmission blocking vaccine candidate.Methods:PCR amplified the dominant antigen epitopes of PyPP5 protein and cloned into pET32a(+)vector.The recombinant PyPP5(rPyPP5)protein was induced by IPTG,purified and immunized mice to obtain anti-PyPP5 serum.The effects of anti-PyPP5 serum on gametocyte exflagellation,ookinete formation and ookinete conversion rate were observed in vitro.Results:Anti-PyPP5 serum was successfully prepared.Anti-PyPP5 serum(at 1∶5 dilution)decreased gametocyte exflagellation,the number of ookinete and the ookinete conversion rate by 29.89%(P<0.05),38.59%(P<0.05)and 64.30%(P<0.01).Conclusion:PyPP5 protein has good antigenicity.Anti-PyPP5 serum can effectively block the transmission of Plasmodium yoelii.