Moser G, Brohn F H, Danforth H D, Nussenzweig R S
J Protozool. 1978 Feb;25(1):119-24. doi: 10.1111/j.1550-7408.1978.tb03881.x.
Sporozoites of rodent malaria, Plasmodium berghei, and simian malaria, Plasmodium knowlesi and Plasmodium cynomolgi, were partially separated from mosquito debris and microbial contaminants by passage of Anopheles material through a DEAE-cellulose column. In addition to eliminating most of the contaminants (80-90%), this simple technic has made it possible to recover rapidly large numbers of viable sporozoites (55-75% yield), which have retained their infectivity, immunogenicity, and capacity to react with known antisera. Mice injected with varying doses of column-purified sporozoites (CS) of P. berghei produced infections which paralleled those seen in the controls. Total protection against challenge with a potentially lethal dose of viable sporozoites was acquired by mice inoculated twice with irradiated CS of P. berghei CS of P. berghei and P. cynomolgi gave positive circumsporozoite precipitation (CSP) reactions, upon inoculation with the respective immune sera. The preservation of the surface antigens of CS was documented by immunofluorescence. It was shown that differences in elution behavior exist among sporozoites of certain species of Plasmodium as well as among sporozoiters of the same species derived from different organs of the mosquito. These results may be attributed to differences in the surface charge of the sporozoites or conditions in sample media. Purified sporozoites obtained by the method described in this report provide an adequate source of parasites for a variety of in vitro studies.
通过将按蚊材料通过二乙氨基乙基纤维素柱，可使啮齿类疟原虫伯氏疟原虫、猴疟原虫诺氏疟原虫和食蟹猴疟原虫的子孢子与蚊子碎片及微生物污染物部分分离。除了去除大部分污染物（80%-90%）外，这种简单技术还能快速回收大量存活的子孢子（回收率55%-75%），这些子孢子保留了其感染性、免疫原性以及与已知抗血清反应的能力。用不同剂量的伯氏疟原虫子孢子柱纯化产物（CS）注射的小鼠产生的感染情况与对照组相似。用伯氏疟原虫的辐照CS两次接种的小鼠获得了针对潜在致死剂量活子孢子攻击的完全保护。伯氏疟原虫和食蟹猴疟原虫的CS接种相应免疫血清后产生阳性环子孢子沉淀（CSP）反应。通过免疫荧光证明了CS表面抗原的保存。结果表明，某些疟原虫物种的子孢子之间以及来自蚊子不同器官的同一物种子孢子之间在洗脱行为上存在差异。这些结果可能归因于子孢子表面电荷或样品介质条件的差异。通过本报告所述方法获得的纯化子孢子为各种体外研究提供了充足的寄生虫来源。
