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兔肺灌洗液中酵母凝集素的特性分析

Characterization of yeast agglutinins in lavage fluid from lungs of rabbits.

作者信息

LaForce F M, Sharrar R G, Arai G

出版信息

J Infect Dis. 1979 Jul;140(1):96-104. doi: 10.1093/infdis/140.1.96.

Abstract

Rabbits were challenged intratracheally with 10(8) Candida albicans or Torulopsis glabrata, and their lungs were lavaged 5, 60, and 120 min later. Initial lavage samples showed significant agglutination of yeasts, followed by the development of larger aggregates in association with alveolar macrophages. To investigate this early agglutination reaction, lungs of normal rabbits were lavaged with heparinized saline, and after alveolar macrophages were discarded, the cell-free lavage fluid was centrifuged at 25,000 g to recover a small, whitish, surface-active pellet (F fraction). The supernatant was concentrated 15-fold by vacuum dialysis (P fraction). When Candida species, T. glabrata, and Saccharomyces cerevisiae were incubated with the F fraction, serial colony counts decreased eight- to 20-fold with every yeast species tested except S. cerevisiae and Candida krusei. Decrease in colony counts was associated with yeast agglutination. The F fraction was further separated by ethanol-ether extraction, and yeast agglutination was seen only in the protein-rich fraction. Further separation of this protein-rich fraction by polyacrylamide gel electrophoresis yielded three bands, one of which, with a molecular size of about 10(4) daltons, agglutinated C. albicans. IgA in the P fraction also agglutinated C. albicans, although not as dramatically as the F fraction.

摘要

用10⁸白色念珠菌或光滑假丝酵母经气管内攻击兔子,5、60和120分钟后对其肺部进行灌洗。最初的灌洗样本显示酵母有明显凝集,随后与肺泡巨噬细胞相关形成更大的聚集体。为研究这种早期凝集反应,用肝素化盐水对正常兔子的肺部进行灌洗,丢弃肺泡巨噬细胞后,将无细胞灌洗液以25000g离心以回收一个小的、白色的、具有表面活性的沉淀(F组分)。通过真空透析将上清液浓缩15倍(P组分)。当白色念珠菌属、光滑假丝酵母和酿酒酵母与F组分一起孵育时,除酿酒酵母和克鲁斯念珠菌外,所测试的每种酵母的系列菌落计数均下降了8至20倍。菌落计数的下降与酵母凝集有关。通过乙醇 - 乙醚萃取进一步分离F组分,仅在富含蛋白质的组分中观察到酵母凝集。通过聚丙烯酰胺凝胶电泳进一步分离该富含蛋白质的组分产生三条带,其中一条分子量约为10⁴道尔顿的带使白色念珠菌凝集。P组分中的IgA也能使白色念珠菌凝集,尽管不如F组分明显。

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