ABSTRACT

Agglutinin-like sequence protein 3 (Als3) is a cell surface glycoprotein of Candida albicans that plays essential roles in the processes of adherence and biofilm formation in vitro. In this study, we focused on the contribution of Als3 to the structure and drug susceptibility of biofilms. The C. albicans wild-type (WT) strain DAY185, the als3Δ/Δ null strain and the als3Δ/Δ + pALS3 complemented strain were used. Colony-forming unit enumeration, crystal violet and cell surface hydrophobicity assays, scanning electron microscopy and confocal laser scanning microscopy coupled with analyses using COMSTAT software were performed to evaluate the biomass and architecture of the biofilms. The detailed architectural analysis showed a significant variation in the biofilm parameters of the als3Δ/Δ biofilms compared with those of the WT biofilms. Fluconazole, miconazole and amphotericin B were selected as the antifungal agents for the antimycotic susceptibility test, and increased susceptibility was found with the ALS3 deletion biofilms. A quantitative real-time polymerase chain reaction analysis showed downregulation of biofilm formation-related genes (ALS1, EFG1, HWP1 and CSH1) and drug resistance-related genes (ERG11, CDR1, CDR2 and MDR1) in the als3Δ/Δ biofilms. We concluded that Als3 contributes to biofilm formation by changing the biofilm architecture and is involved in the antifungal resistance of C. albicans biofilms.

中文翻译：

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凝集素样序列蛋白 3 (Als3) 在白色念珠菌生物膜结构和抗真菌性中的作用

摘要

凝集素样序列蛋白 3 (Als3) 是白色念珠菌的细胞表面糖蛋白，在体外粘附和生物膜形成过程中起重要作用。在这项研究中，我们专注于 Als3 对生物膜结构和药物敏感性的贡献。的白色念珠菌的野生型（WT）应变DAY185，所述ALS3 Δ/Δ空菌株和ALS3 Δ/Δ+ P ALS3使用补充菌株。进行菌落形成单位计数、结晶紫和细胞表面疏水性测定、扫描电子显微镜和共聚焦激光扫描显微镜以及使用 COMSTAT 软件的分析，以评估生物膜的生物量和结构。详细的结构分析表明，与 WT 生物膜相比，als3 Δ/Δ 生物膜的生物膜参数存在显着差异。氟康唑、咪康唑和两性霉素 B 被选为抗真菌药敏试验的抗真菌剂，发现ALS3缺失生物膜的易感性增加。定量实时聚合酶链反应分析显示生物膜形成相关基因（ALS1、EFG1、HWP1和CSH1）和als3 Δ/Δ 生物膜中的耐药相关基因（ERG11、CDR1、CDR2和MDR1）。我们得出结论，Als3 通过改变生物膜结构有助于生物膜形成，并参与白色念珠菌生物膜的抗真菌抗性。