Department of Cell Biology, Center for Research and Advanced Studies, Av. Instituto Politécnico Nacional 2508, Zacatenco 07360, México D.F., City 07360, México.
Can J Microbiol. 2010 Dec;56(12):987-95. doi: 10.1139/W10-088.
Trophozoites of Entamoeba histolytica HM-1:IMSS become less virulent after long-term maintenance in axenic cultures. The factors responsible for the loss of virulence during in vitro cultivation remain unclear. However, it is known that in vitro cultivation of amoeba in culture medium supplemented with cholesterol restores their virulence. In this study, we analyzed the effect of adding phosphatidylcholine-cholesterol (PC-Chol) liposomes to the culture medium and evaluated the effect of this lipid on various biochemical and biological functions of E. histolytica HM-1:IMSS in terms of its virulence. The addition of PC-Chol liposomes to the culture medium maintained the virulence of these parasites against hamster liver at the same level as the original virulent E. histolytica strain, even though these amoebae were maintained without passage through hamster liver for 18 months. The trophozoites also showed increased endocytosis, erythrophagocytosis, and carbohydrate residue expression on the amoebic surface. Protease activities were also modified by the presence of cholesterol in the culture medium. These findings indicate the capacity of cholesterol to preserve amoeba virulence and provide an alternative method for the maintenance of virulent E. histolytica trophozoites without the need for in vivo procedures.
溶组织内阿米巴滋养体 HM-1:IMSS 在长期的无基质培养中其毒力降低。导致体外培养过程中丧失毒力的因素尚不清楚。然而,已知在含有胆固醇的培养基中体外培养滋养体可恢复其毒力。在本研究中,我们分析了向培养基中添加磷脂酰胆碱-胆固醇(PC-Chol)脂质体的效果,并评估了该脂质对溶组织内阿米巴 HM-1:IMSS 各种生化和生物学功能的影响,包括其毒力。向培养基中添加 PC-Chol 脂质体可使这些寄生虫对仓鼠肝脏的毒力保持在与原始毒力溶组织内阿米巴株相同的水平,即使这些阿米巴在 18 个月内没有通过仓鼠肝脏传代。滋养体的内吞作用、红细胞吞噬作用和阿米巴表面碳水化合物残留的表达也增加。培养基中胆固醇的存在还改变了蛋白酶的活性。这些发现表明胆固醇有保持阿米巴毒力的能力,并提供了一种替代方法来维持具有毒力的溶组织内阿米巴滋养体,而无需进行体内操作。