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利用单克隆抗体分析胎儿三毛滴虫对哺乳动物细胞的黏附及细胞毒性

Analysis of adhesion and cytotoxicity of Tritrichomonas foetus to mammalian cells by use of monoclonal antibodies.

作者信息

Burgess D E, McDonald C M

机构信息

Veterinary Molecular Biology, Montana State University, Bozeman 59717.

出版信息

Infect Immun. 1992 Oct;60(10):4253-9. doi: 10.1128/iai.60.10.4253-4259.1992.

Abstract

The relationship of Tritrichomonas foetus adhesion to mammalian cells and cytotoxicity to these targets was investigated. High-adherence and low-adherence lines of T. foetus, derived by repeated adhesion to HeLa cells, showed high and low cytotoxicity, respectively, to HeLa cells. When parasites were separated from targets by membranes (0.4-microns pore size), no cytotoxicity was detectable. Monoclonal antibodies elicited against T. foetus that lowered adhesion also lowered parasite-mediated cytotoxicity. Flow cytometry experiments revealed that the levels of an adhesion- and cytotoxicity-blocking antibody bound to the surface of high-adherence clones of T. foetus were higher than those in low-adherence clones. Western blots of parasite extracts separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis were probed with an anti-T. foetus antibody. A molecule with a molecular weight of approximately 190,000 composed of subunits with molecular weights of approximately 140,000 to 150,000 and approximately 65,000 was identified. Immunoprecipitation experiments with metabolically labeled T. foetus and the same antibody confirmed that similar subunits were synthesized by the parasite. These results indicate that adhesion of T. foetus to mammalian cells is an important step in cytotoxic damage of these targets and that a surface adhesin on the parasite is involved in the adhesion mechanism.

摘要

研究了胎儿三毛滴虫对哺乳动物细胞的黏附作用及其对这些靶标的细胞毒性之间的关系。通过反复黏附于HeLa细胞而获得的胎儿三毛滴虫高黏附性和低黏附性品系,分别对HeLa细胞表现出高细胞毒性和低细胞毒性。当寄生虫通过膜(孔径0.4微米)与靶标分离时,未检测到细胞毒性。针对胎儿三毛滴虫产生的降低黏附作用的单克隆抗体也降低了寄生虫介导的细胞毒性。流式细胞术实验显示,与胎儿三毛滴虫高黏附性克隆表面结合的一种黏附及细胞毒性阻断抗体的水平高于低黏附性克隆。用抗胎儿三毛滴虫抗体对经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离的寄生虫提取物进行蛋白质免疫印迹分析。鉴定出一种分子量约为190,000的分子,它由分子量约为140,000至150,000和约65,000的亚基组成。用代谢标记的胎儿三毛滴虫和相同抗体进行的免疫沉淀实验证实,寄生虫合成了相似的亚基。这些结果表明,胎儿三毛滴虫对哺乳动物细胞的黏附是这些靶标细胞毒性损伤的一个重要步骤,并且寄生虫表面的一种黏附素参与了黏附机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b7d/257460/ecb720e1eeed/iai00034-0318-a.jpg

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