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首页> 外文期刊>Journal of Clinical Microbiology >Isolation of a cell-detaching factor of Trichomonas vaginalis.
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Isolation of a cell-detaching factor of Trichomonas vaginalis.

机译:分离阴道毛滴虫的细胞分离因子。

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The pathogenetic role of soluble products of Trichomonas vaginalis growth in culture is controversial. To evaluate this role, T. vaginalis was grown in broth and cell culture and the cell-free filtrate was applied to fresh cell culture monolayers. When adjusted to pH 6.5, filtrates obtained from 22-h culture growth totally disrupted McCoy, HEp-2, human foreskin fibroblast, and Chinese hamster ovary cell monolayers within 6 h. These detached cells remained greater than 90% viable. This cell-detaching factor (CDF) was heat and acid labile, with a pH optimum of 6.5. CDF has trypsinlike activity which disrupts monolayer cells, but cells do not die if the pH is controlled. CDF was purified by ethanol precipitation, ammonium sulfate fractionation, and ion-exchange and gel filtration column chromatography. A 200,000-molecular-weight glycoprotein which was also immunogenic by immunoblot with human sera reactive to T. vaginalis was isolated in this manner. This confirms the presence of a specific soluble CDF derived from T. vaginalis whose application may be important as a diagnostic tool and in further studies of pathogenesis.
机译:阴道毛滴虫生长中的可溶性产物在培养中的致病作用引起争议。为了评估这一作用,将阴道锥虫在肉汤中培养并进行细胞培养,并将无细胞滤液应用于新鲜的细胞培养单层。调节至pH 6.5时,在22小时的培养过程中得到的滤液在6小时内完全破坏了McCoy,HEp-2,人包皮成纤维细胞和中国仓鼠卵巢单层细胞。这些分离的细胞保持大于90%的存活率。该细胞分离因子(CDF)是热和酸不稳定的,最适pH为6.5。 CDF具有类胰蛋白酶活性,可破坏单层细胞,但如果控制pH值,细胞不会死亡。通过乙醇沉淀,硫酸铵分级分离,离子交换和凝胶过滤柱色谱法纯化CDF。以这种方式分离了200,000分子量的糖蛋白,其也通过免疫印迹用对阴道毛滴虫具有反应性的人血清进行免疫原化。这证实了存在于阴道锥虫的特定可溶性CDF,其应用可能作为诊断工具和进一步的发病机理研究很重要。

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