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Isolation of a Soluble Component of Plasmodium berghei Which Induces Immunity in Rats

机译：分离可诱导大鼠免疫力的伯氏疟原虫可溶性成分

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Soluble material was obtained from sonically freed plasmodiae by three procedures. Two procedures, cryo-impacting and freeze-thawing, were evaluated for their ability to disrupt the parasites and release soluble material. The soluble materials obtained by these procedures were compared to materials washed from the surfaces of sonically freed parasites. Between 35 and 40% of the total parasite protein was solubilized by freeze-thawing or cryo-impacting. One cycle of freeze-thawing released nearly as much protein as could be released by this method, and additional cycles of freeze-thawing had little additional effect. Cryo-impacting solubilized only a small amount of protein in addition to that which was released by the cycle of freeze-thawing inherent in the procedure. Reductions in the packed cell volume of the material remaining after freeze-thawing or cryo-impacting indicate that the insoluble fragments are broken into smaller pieces as treatment is extended. Electron microscopy of 30-s cryo-impacted and three-times freeze-thawed parasites revealed membrane fragments similar in appearance. Patterns obtained by polyacrylamide gel electrophoresis of the soluble material from freeze-thawed and cryo-impacted parasites were also similar, and approximately 13 protein bands were demonstrated. The material washed from the surfaces of the free parasites, on the other hand, resolved into only two to four major bands on the gel columns. In immunization studies, the soluble and insoluble fractions obtained by freeze-thawing or cryo-impacting and the material washed from the surfaces of the parasites all stimulated a protective immune response. On the basis of the amount of protein required to stimulate roughly comparable immunity, the soluble fraction obtained by freeze-thawing or cryo-impacting free parasites was about twice as potent an immunogen as was the insoluble fraction. The material obtained by gentle washing of the freed parasites was approximately 20 times as potent an immunogen as were the freed parasites and about 7 times as potent as the soluble material obtained by freeze-thawing or cryo-impacting.
机译：通过三种方法从超声释放的疟原虫中获得可溶性物质。评价了两种方法，即冷冻冲击法和冷冻-解冻法，它们破坏寄生虫并释放可溶性物质的能力。将通过这些程序获得的可溶物质与从超声释放的寄生虫表面洗涤的物质进行比较。通过冷冻-解冻或冷冻冲击，可溶解总寄生蛋白的35％至40％。冷冻解冻的一个循环释放的蛋白质与通过此方法释放的蛋白质几乎相同，另外的冷冻解冻循环几乎没有其他效果。除了通过该程序固有的冻融循环释放的蛋白质以外，低温撞击仅溶解了少量的蛋白质。冻融或冷冻冲击后残留材料的填充细胞体积减少表明，随着处理时间的延长，不溶性碎片会破碎成较小的碎片。 30 s冷冻冲击和三倍冻融的寄生虫的电子显微镜显示出膜碎片外观相似。通过聚丙烯酰胺凝胶电泳从冻融和冷冻影响的寄生虫中获得的可溶物质的模式也相似，并且证实了大约13条蛋白带。另一方面，从游离寄生虫表面洗掉的物质在凝胶柱上仅分解为两到四个主要谱带。在免疫研究中，通过冻融或冷冻冲击获得的可溶和不溶级分以及从寄生虫表面洗涤的物质均刺激了保护性免疫反应。根据刺激大致相当的免疫力所需的蛋白质量，通过冻融或冷冻作用的游离寄生虫获得的可溶级分的免疫原效力约为不溶级分的两倍。通过温和洗涤游离的寄生虫而获得的材料，其免疫原的效力是游离的寄生虫的约20倍，而其有效力是通过冻融或冷冻冲击获得的可溶性材料的约7倍。

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