Alviano Daniela S, Kneipp Lucimar F, Lopes Angela H, Travassos Luiz R, Meyer-Fernandes José R, Rodrigues Marcio L, Alviano Celuta S
Instituto de Microbiologia Professor Paulo de Góes, Departamento de Microbiologia Geral, Universidade Federal do Rio de Janeiro, Cidade, Rio de Janeiro, Brazil.
Res Microbiol. 2003 Dec;154(10):689-95. doi: 10.1016/j.resmic.2003.09.002.
Platelet-activating factor (PAF) has been shown to induce the differentiation of several cell types. In this work, we evaluated the effects of PAF on the formation of sclerotic cells of Fonsecaea pedrosoi, the major causative agent of chromoblastomycosis. Cell differentiation was evaluated by light and electron microscopy, which showed that treatment of mycelial forms with PAF results in the generation of sclerotic bodies with typical morphological characteristics. Biochemical features of PAF-induced sclerotic cells were also analyzed and compared with those from sclerotic forms induced by propranolol, a previously described differentiating agent of F. pedrosoi. Chemical analyses of lipid and carbohydrate components from PAF- or propranolol-induced sclerotic bodies revealed that palmitic, stearic, oleic and linoleic acids were the major fatty acid components, while glucose, mannose, galactose and rhamnose were detected as the principal sugar constituents in these cells. Surface carbohydrate components of PAF- and propranolol-induced sclerotic cells were also evaluated, by flow cytometry analysis with twelve different lectins. The profile of reactivity of PAF- or propranolol-induced fungal cells with lectins was also very similar. Hydrolysis of the synthetic substrate p-nitrophenylphosphate by fungal cells demonstrated that the addition of PAF or propranolol to the mycelial cultures similarly promotes a significant increase in ecto-phosphatase activity. These results indicate that the differentiation of F. pedrosoi mycelial cells induced by PAF generates authentic sclerotic forms, as confirmed by the analysis of morphological and biochemical attributes. Since PAF is synthesized in normal conditions by the human host, these observations may have a correlation with the differentiation of F. pedrosoi in vivo.
血小板活化因子(PAF)已被证明可诱导多种细胞类型的分化。在本研究中,我们评估了PAF对裴氏着色真菌(chromoblastomycosis的主要病原体)硬化细胞形成的影响。通过光学显微镜和电子显微镜评估细胞分化,结果表明用PAF处理菌丝体形式可产生具有典型形态特征的硬化体。还分析了PAF诱导的硬化细胞的生化特征,并与普萘洛尔(一种先前描述的裴氏着色真菌分化剂)诱导的硬化形式的生化特征进行了比较。对PAF或普萘洛尔诱导的硬化体的脂质和碳水化合物成分进行化学分析,结果显示棕榈酸、硬脂酸、油酸和亚油酸是主要脂肪酸成分,而葡萄糖、甘露糖、半乳糖和鼠李糖是这些细胞中的主要糖类成分。还通过用十二种不同凝集素进行流式细胞术分析,评估了PAF和普萘洛尔诱导的硬化细胞的表面碳水化合物成分。PAF或普萘洛尔诱导的真菌细胞与凝集素的反应性谱也非常相似。真菌细胞对合成底物对硝基苯磷酸酯的水解表明,向菌丝体培养物中添加PAF或普萘洛尔同样会显著促进胞外磷酸酶活性的增加。这些结果表明,通过形态学和生化属性分析证实,PAF诱导的裴氏着色真菌菌丝体细胞分化产生了真正的硬化形式。由于PAF在正常情况下由人类宿主合成,这些观察结果可能与裴氏着色真菌在体内的分化有关。