Differential expression of secreted aspartyl proteinases in a model of human oral candidosis and in patient samples from the oral cavity
Differential expression of secreted aspartyl proteinases in a model of human oral candidosis and in patient samples from the oral cavity
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DOI：
10.1046/j.1365-2958.1998.00957.x
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发表时间：
1998-07-01
影响因子：
3.6
通讯作者：
Hube, B
中科院分区：
文献类型：
--
作者：
Schaller, M;Schäfer, W;Hube, B
白色念珠菌是人类的一种机会性病原体，它分泌与毒力相关的分泌性天冬氨酸蛋白酶(SAPS)。我们通过逆转录聚合酶链式反应(RT-PCR)检测了SAP基因家族中七个已知成员在(I)基于重组人上皮(RHE)的口腔念珠菌病体外模型和(Ii)口腔念珠菌病患者的临床样本中的mRNA表达的时间调节。SAP1和SAP3转录本在接种RHE后42h首次检测到，同时在光镜下观察到上皮细胞有轻微的形态变化。SAP6的表达在6h后出现，同时伴有一些感染念珠菌细胞的生殖管形成和严重的上皮组织病变。SAP2和SAP8的RT-PCR产物在感染后60h首次被检测到，而SAP4和SAPS的转录本从未被检测到。因此，随着RHE损伤的增加，SAP表达的时间顺序为SAP1和SAP3>；SAP6>；SAP2和SAP8。在蛋白质水平上，使用针对基因产物Sap1-3的抗SAP鼠单抗，在免疫电子显微镜下，在白念珠菌酵母细胞和上皮细胞中发现了SAP抗原。用RT-PCR方法检测口腔念珠菌病患者标本中SAP1-3和SAP1-6的表达。我们的结果表明，实验性和临床性口腔念珠菌病的发病机制与SAP基因表达的差异性和时间性调节有关。
Candida albicans, an opportunistic pathogen in humans, secretes secretory aspartyl proteinases (Saps), which have been correlated with virulence. We examined the temporal regulation of the mRNA expression of seven known members of the SAP gene family by reverse transcription polymerase chain reaction (RT-PCR) in (i) an in vitro model of oral candidosis based an reconstituted human epithelium (RHE); and (ii) clinical samples from patients with oral candidosis. SAP1 and SAP3 transcripts were first detected 42h after inoculation of RHE, while at the same time, slight morphological alterations in the epithelium were documented by light microscopy. SAP6 expression occurred 6h later concomitantly with germ tube formation of some infecting Candida cells and severe lesions of the epithelial tissue. SAP2 and SAP8 RT-PCR products were first detected 60 h after infection, while SAP4 and SAPS transcripts were never discovered. Thus a temporal progression of SAP expression in the order SAP1 and SAP3> SAP6> SAP2 and SAP8 was observed at the same time as increasing RHE damage occurred. At the protein level, Sap antigen was found within the C. albicans yeast cells and the epithelial cells by immunoelectron microscopy using an anti-Sap murine monoclonal antibody directed against the gene products Sap1-3. Expression of SAP1-3 and 6 was also detected by RT-PCR in samples from patients suffering from oral candidosis. Our results suggest that the pathogenesis of experimental and clinical oral candidosis is associated with the differential and temporal regulation of SAP gene expression.