Rochetti Victor Pereira, Rollin-Pinheiro Rodrigo, de Oliveira Evely Bertulino, Xisto Mariana Ingrid Dutra da Silva, Barreto-Bergter Eliana
Laboratório de Química Biológica de Microrganismos, Departamento de Microbiologia Geral, Instituto de Microbiologia Paulo de Góes, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro 21941-902, Brazil.
J Fungi (Basel). 2020 Dec 8;6(4):345. doi: 10.3390/jof6040345.
Infections caused by species present a wide range of clinical manifestations, from superficial to disseminated, especially in immunocompromised patients. Glucosylceramides (GlcCer) are glycosphingolipids found on the fungal cell surface and play an important role in growth and pathogenicity processes in different fungi. The present study aimed to evaluate the structure of GlcCer and its role during growth in two isolates. Purified GlcCer from both isolates were obtained and its chemical structure identified by mass spectrometry. Using ELISA and immunofluorescence techniques it was observed that germination and NaOH-treatment of conidia favor GlcCer exposure. Monoclonal anti-GlcCer antibody reduced germination when cultivated with the inhibitor of melanin synthesis tricyclazole and also reduced germ tube length of conidia, both cultivated or not with tricyclazole. It was also demonstrated that anti-GlcCer altered lipid rafts organization, as shown by using the fluorescent stain filipin, but did not affect the susceptibility of the cell surface to damaging agents. Anti-GlcCer reduced total biomass and viability in biofilms formed on polystyrene plates. In the presence of anti-GlcCer, germinated conidia and biofilms could not adhere to polystyrene with the same efficacy as control cells. These results highlight the relevance of GlcCer in growth processes of .
由该菌种引起的感染表现出广泛的临床表现，从浅表感染到播散性感染，尤其是在免疫功能低下的患者中。葡糖神经酰胺（GlcCer）是在真菌细胞表面发现的糖鞘脂，在不同真菌的生长和致病过程中起重要作用。本研究旨在评估GlcCer的结构及其在两种分离株生长过程中的作用。从两种分离株中获得了纯化的GlcCer，并通过质谱鉴定了其化学结构。使用酶联免疫吸附测定（ELISA）和免疫荧光技术观察到，分生孢子的萌发和氢氧化钠处理有利于GlcCer的暴露。当与黑色素合成抑制剂三环唑一起培养时，单克隆抗GlcCer抗体降低了萌发率，并且在有或没有三环唑培养的情况下，也降低了分生孢子的芽管长度。还证明了抗GlcCer改变了脂筏的组织，如使用荧光染料制霉菌素所显示的那样，但不影响细胞表面对损伤剂的敏感性。抗GlcCer降低了在聚苯乙烯平板上形成的生物膜中的总生物量和活力。在存在抗GlcCer的情况下，萌发的分生孢子和生物膜不能像对照细胞那样有效地粘附在聚苯乙烯上。这些结果突出了GlcCer在该菌种生长过程中的相关性。
