【摘要】 目的从6B、9V、14、19F型肺炎球菌培养物中纯化肺炎球菌溶血素(PLY),并检测其免疫原性和保护作用。方法采用自溶、硫酸铵沉淀及三步柱层析法纯化PLY,并检测PLY免疫NIH小鼠后血清抗体滴度及其保护效果。结果纯化的6B、9V、14、19F型PLY纯度依次为83.4%、99.2%、70.1%和76.6%;9V型PLY表观相对分子质量为52840,与文献报道的53000相一致。从4种血清型肺炎球菌中纯化的PLY均能诱导小鼠产生高且稳定持久的抗体滴度。各免疫组小鼠分别感染6B、9V、14、19F型肺炎球菌,存活率均显著高于对照组。结论4种血清型PLY均具有良好的免疫原性和保护作用。更多还原

【Abstract】 Objective To purify pneumolysin (PLY) of types 6 B,9 V,14,and 19 F from the culture of pneumococcus of the corresponding types and study their immunogenicity and protective effect.Methods Purify PLY from the culture of pneumococcus by autolysis,ammonium sulfate fractionation and ion exchang,gel filtration and affiniuy chromatography.Immunize NIH mice with the purified PLY and determine the serum antibody titer.Challenge the immunized mice with pneumococcus to evaluate the protective effect of PLY.Results The purities of purified PLY of types 6 B,9 V,14 and 19 F were 83.4%,99.2%,70.1% and 76.6% respectively.The apparent relative molecular mass of PLY of type 9 V was 52 840,which was consistent with that reported (53 000).All the 4 types of PLY induced high and persistent antibody titers in mice.After challenge with pneumococcus of the corresponding types,the survival rates of immunized mice were significantly higher than those of control.Conclusion All the 4 types of purified PLY showed good immunogenicity and protecive effect.更多还原