Blasi E, Pitzurra L, Chimienti A R, Mazzolla R, Puliti M, Barluzzi R, Bistoni F
Department of Experimental Medicine and Biochemical Sciences, University of Perugia, Italy.
Infect Immun. 1995 Apr;63(4):1253-7. doi: 10.1128/iai.63.4.1253-1257.1995.
The dimorphic transition of Candida albicans from the yeast (Y-Candida) to the hyphal (H-Candida) form is a complex event whose relevance in fungal pathogenicity is still poorly understood. Using a cloned macrophage (M phi) cell line (ANA-1), we have previously shown that a M phi can discriminate between the two fungal forms, eliciting different secretory responses. In the present study, we investigated the susceptibility of Y-Candida and H-Candida to M phi proteolytic activity. In particular, sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot (immunoblot) techniques were employed to analyze the patterns of lyticase proteinaceous extracts from cell walls of Y-Candida and H-Candida which had been unexposed or exposed to ANA-1 M phis for 3 h. Silver staining allowed detection of a complex protein pattern in both forms of C. albicans, qualitatively and quantitatively differing from each other, mainly at molecular masses below 106 kDa. Western blot staining with anti-C. albicans mannan antibodies and convalescent-phase sera of mice previously infected systemically or intracerebrally with C. albicans showed that, after contact with M phis, Y-Candida but not H-Candida proteinaceous cell wall components are profoundly modified, with substantial reduction and/or disappearance of many bands. Our experimental approach provides initial insights into the differential susceptibility of Y-Candida and H-Candida to the proteolytic activity of M phis.
白色念珠菌从酵母形态(Y-念珠菌)向菌丝形态(H-念珠菌)的双态转变是一个复杂的过程,其在真菌致病性中的作用仍知之甚少。我们之前利用克隆的巨噬细胞(M phi)细胞系(ANA-1)表明,巨噬细胞能够区分这两种真菌形态,并引发不同的分泌反应。在本研究中,我们调查了Y-念珠菌和H-念珠菌对巨噬细胞蛋白水解活性的敏感性。具体而言,采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和蛋白质印迹(免疫印迹)技术,分析未接触或接触ANA-1巨噬细胞3小时的Y-念珠菌和H-念珠菌细胞壁溶菌酶蛋白提取物的模式。银染法能够检测到白色念珠菌两种形态中复杂的蛋白质模式,两者在定性和定量上均存在差异,主要在分子量低于106 kDa的区域。用抗白色念珠菌甘露聚糖抗体和先前经白色念珠菌全身或脑内感染的小鼠恢复期血清进行蛋白质印迹染色表明,与巨噬细胞接触后,Y-念珠菌而非H-念珠菌的细胞壁蛋白质成分发生了显著改变,许多条带大幅减少和/或消失。我们的实验方法为Y-念珠菌和H-念珠菌对巨噬细胞蛋白水解活性的不同敏感性提供了初步见解。