Burgess D E, Knoblock K F, Daugherty T, Robertson N P
Veterinary Molecular Biology Laboratory, Montana State University, Bozeman 59717.
Infect Immun. 1990 Nov;58(11):3627-32. doi: 10.1128/iai.58.11.3627-3632.1990.
Geographically distinct lines of Tritrichomonas foetus were assayed for their ability to cause cytotoxicity in nucleated mammalian cells and lysis of bovine erythrocytes. T. foetus was highly cytotoxic toward a human cervical cell line (HeLa) and early bovine lymphosarcoma (BL-3) but displayed low levels of cytotoxicity against African green monkey kidney (Vero) cells. In addition to variation in the extent of cytotoxicity toward different targets, differences in the levels of cytotoxicity in the same nucleated target occurred with different parasite lines. Whole T. foetus, unfractionated whole-cell extracts, and parasite-conditioned medium (RPMI 1640 without serum) all caused lysis of bovine erythrocytes. Lytic activity in the conditioned medium was substantially reduced by repeated freezing and thawing or heating to 90 degrees C for 30 min. Damage of mammalian target cells by live T. foetus could be reduced by the presence of protease inhibitors; however, such inhibitors did not diminish the lytic effects of conditioned medium. These results suggested that proteolytic enzymes were necessary for the lytic mechanism of the live parasites but were not required once lytic factors were released into the parasite-conditioned medium. They further suggested that the lytic molecules were either proteins or had proteinaceous components.
对来自不同地理区域的胎儿三毛滴虫株系进行了检测,以评估它们在有核哺乳动物细胞中引起细胞毒性以及裂解牛红细胞的能力。胎儿三毛滴虫对人宫颈癌细胞系(HeLa)和早期牛淋巴细胞瘤(BL - 3)具有高度细胞毒性,但对非洲绿猴肾(Vero)细胞的细胞毒性水平较低。除了对不同靶标的细胞毒性程度存在差异外,不同寄生虫株系对同一有核靶标的细胞毒性水平也有所不同。完整的胎儿三毛滴虫、未分级的全细胞提取物以及寄生虫条件培养基(不含血清的RPMI 1640)均可引起牛红细胞的裂解。条件培养基中的裂解活性通过反复冻融或加热至90℃ 30分钟而大幅降低。蛋白酶抑制剂的存在可降低活的胎儿三毛滴虫对哺乳动物靶细胞的损伤;然而,此类抑制剂并未减弱条件培养基的裂解作用。这些结果表明,蛋白水解酶对于活寄生虫的裂解机制是必需的,但一旦裂解因子释放到寄生虫条件培养基中则不再需要。它们进一步表明,裂解分子要么是蛋白质,要么具有蛋白质成分。