CEB - Centre of Biological Engineering, LIBRO - Laboratório de Investigação em Biofilmes Rosário Oliveira, University of Minho, 4710-057 Braga, Portugal.
Tissue Engineering & Reparative Dentistry, School of Dentistry, Cardiff University, Cardiff CF14 4XY, United Kingdom.
J Infect. 2014 Oct;69(4):396-407. doi: 10.1016/j.jinf.2014.06.002. Epub 2014 Jun 9.
The principal aim of this study was to investigate the in vitro co-infection of a reconstituted human vaginal epithelium (RHVE) by Candida albicans and Candida glabrata.
The ability of both species to invade and colonise the RHVE was examined using species-specific peptide nucleic acid (PNA) probe hybridisation, confocal laser scanning microscopy (CLSM) and a novel qRT-PCR protocol for Candida quantification in the tissues. RHVE damage was evaluated by measuring lactate dehydrogenase (LDH) activity. Candida virulence gene expression (HWP1, ALS, EPA, PLB, PLD and SAP) was evaluated by quantitative RT-PCR.
The results showed that whilst both species induced damage to the RHVE, this was notably less with C. glabrata. Interestingly, there was a significant increase in C. glabrata RHVE colonisation and invasiveness when it was added to the tissue with C. albicans. The extent of RHVE damage caused by the two species appeared to be primarily dependent on the process of invasion. Of the virulence genes assayed, HWP1, PLD1 and ALS3 were deemed to be most associated with pathogenicity in the model.
For the first time, we have demonstrated that the RHVE model coupled with specific tools of analysis, allows assessment of Candida colonisation and invasion in single and co-infection. Using this model we have demonstrated that C. albicans enhanced C. glabrata colonisation, invasion and tissue damage, which was also evidenced by the expression of virulence genes.
本研究的主要目的是研究重组人阴道上皮（RHVE）中白色念珠菌和光滑念珠菌的体外共感染。
使用种特异性肽核酸（PNA）探针杂交、共聚焦激光扫描显微镜（CLSM）和用于组织中念珠菌定量的新型 qRT-PCR 方案，检查两种物种入侵和定殖 RHVE 的能力。通过测量乳酸脱氢酶（LDH）活性评估 RHVE 损伤。通过定量 RT-PCR 评估念珠菌毒力基因表达（HWP1、ALS、EPA、PLB、PLD 和 SAP）。
结果表明，虽然两种物种都导致 RHVE 损伤，但 C. glabrata 引起的损伤明显较少。有趣的是，当 C. glabrata 添加到与 C. albicans 共培养的组织中时，其 RHVE 定植和侵袭性显著增加。两种物种引起的 RHVE 损伤程度似乎主要取决于侵袭过程。在所检测的毒力基因中，HWP1、PLD1 和 ALS3 被认为与该模型中的致病性最相关。
我们首次证明，RHVE 模型结合特定的分析工具，允许评估念珠菌在单一和共感染中的定植和入侵。使用该模型，我们证明了 C. albicans 增强了 C. glabrata 的定植、侵袭和组织损伤，这也可以通过表达毒力基因来证明。
