Borg-von Zepelin M, Eucker J, Rüchel R
Abteilung Medizinische Mikrobiologie, Hygiene-Institut der Universität Göttingen, BR Deutschland.
Mycoses. 1997;40 Suppl 1:64-72. doi: 10.1111/j.1439-0507.1997.tb00544.x.
The influence of the heterologous acid secretory Candida proteinases on the adherence of the non-proteinase secreting strain of C. tropicalis DSM 4959 to epitheloid cells (vero line) was examined. The proteinases of the following Candida strains were used: C. albicans ATCC 10261 (serotype A), C. albicans ATCC 48867 (serotype B), C. tropicalis DSM 4238. The assays were performed with the previously described in-vitro-adherence test [1] using the following principle steps: Candida proteinases and C. tropicalis blastoconidia were incubated with verocells in microtest plates in phosphate-buffer in the range of pH 4.0 to pH 7.0. Adherent Candida cells were detected according to Filler et al. [2] with anti-Candida-mannoprotein antibodies and a secondary anti-rabbit-peroxidase conjugate. Compared to controls with denaturated proteinases, the photometric evaluation of adherent C. tropicalis cells showed, under optimal conditions, an augmentation of the adherence due to the Candida proteinases of about 50%. The optimum of this adherence augmentation was in the range of pH 5.5 which is outside the general activity optimum of Candida proteinases (pH 3). The degree of purity of these proteinases had no marked influence on the adherence. The specificity of the proteinase dependent adherence augmentation could be demonstrated with the enzyme inhibitor Pepstatin A. C. tropicalis blastoconidia supplemented by pepstatin A and active Candida proteinase adhered in the same range as with denaturated proteinases in control tests. Our results suggest a function of Candida proteinases in the adherence process of blastoconidia to epithelia.
研究了异源酸性分泌型念珠菌蛋白酶对热带念珠菌DSM 4959非蛋白酶分泌菌株与上皮样细胞（vero细胞系）黏附的影响。使用了以下念珠菌菌株的蛋白酶：白色念珠菌ATCC 10261（血清型A）、白色念珠菌ATCC 48867（血清型B）、热带念珠菌DSM 4238。采用先前描述的体外黏附试验[1]，按照以下主要步骤进行测定：将念珠菌蛋白酶和热带念珠菌芽生孢子在pH 4.0至pH 7.0的磷酸盐缓冲液中于微量滴定板中与vero细胞一起孵育。根据Filler等人[2]的方法，用抗念珠菌甘露糖蛋白抗体和抗兔过氧化物酶二级偶联物检测黏附的念珠菌细胞。与变性蛋白酶对照组相比，在最佳条件下，对黏附的热带念珠菌细胞进行光度评估显示，由于念珠菌蛋白酶，黏附增加了约50%。这种黏附增加的最佳pH范围是5.5，这超出了念珠菌蛋白酶的一般活性最佳pH值（pH 3）。这些蛋白酶的纯度对黏附没有明显影响。蛋白酶依赖性黏附增加的特异性可以用酶抑制剂胃蛋白酶抑制剂A来证明。在对照试验中，添加胃蛋白酶抑制剂A和活性念珠菌蛋白酶的热带念珠菌芽生孢子的黏附情况与变性蛋白酶的黏附情况相同。我们的结果表明念珠菌蛋白酶在芽生孢子与上皮细胞的黏附过程中起作用。
