Pinto Lysianne, Granja Luiz Fernando Zmetek, Almeida Mariana Amorim de, Alviano Daniela Sales, Silva Maria Helena da, Ejzemberg Regina, Rozental Sonia, Alviano Celuta Sales
Universidade Federal do Rio de Janeiro, Instituto de Microbiologia Professor Paulo de Góes, Rio de Janeiro, RJ, Brasil.
Universidade Federal do Rio de Janeiro, Instituto de Macromoléculas Professora Eloisa Mano, Rio de Janeiro, RJ, Brasil.
Mem Inst Oswaldo Cruz. 2018 Jun 25;113(8):e180120. doi: 10.1590/0074-02760180120.
Melanin production has been associated with virulence in various pathogenic fungi, including Fonsecaea pedrosoi, the major etiological agent for chromoblastomycosis, a subcutaneous fungal disease that occurs in South America.
The aim of this study was to evaluate the effects of acid-basic extracted F. pedrosoi melanin particles and fungal cell ghosts obtained by Novozym 234 treatment on their ability to activate the human complement system.
The ability of melanin particles and fungal cell ghosts to activate the human complement system was evaluated by complement consumption, immunofluorescence, and enzyme-linked immunosorbent assay (ELISA).
Unsensitised melanin particles and melanin ghosts presented complement consumption of 82.67 ± 2.08% and 96.04 ± 1.13%, respectively. Immunofluorescence assays revealed intense deposition of the C3 and C4 fragments on the surface of melanin particles and ghosts extracted from F. pedrosoi. Deposition of the C3, C4, and C5 fragments onto melanin samples and zymosan was confirmed by ELISA. Deposition of small amounts of C1q and C9 onto melanin samples and zymosan was detected by ELISA.
Fonsecaea pedrosoi melanin particles and fungal cell ghosts activated the complement system mainly through an alternative pathway.
黑色素的产生与多种致病真菌的毒力相关,包括裴氏着色真菌,它是南美洲发生的一种皮下真菌病——着色芽生菌病的主要病原体。
本研究旨在评估酸碱提取的裴氏着色真菌黑色素颗粒和经诺维信234处理获得的真菌细胞空壳对其激活人类补体系统能力的影响。
通过补体消耗、免疫荧光和酶联免疫吸附测定(ELISA)评估黑色素颗粒和真菌细胞空壳激活人类补体系统的能力。
未致敏的黑色素颗粒和黑色素空壳的补体消耗分别为82.67±2.08%和96.04±1.13%。免疫荧光分析显示,C3和C4片段在从裴氏着色真菌提取的黑色素颗粒和空壳表面有强烈沉积。ELISA证实了C3、C4和C5片段在黑色素样品和酵母聚糖上的沉积。ELISA检测到少量C1q和C9在黑色素样品和酵母聚糖上的沉积。
裴氏着色真菌黑色素颗粒和真菌细胞空壳主要通过替代途径激活补体系统。