Rane Hallie S, Hardison Sarah, Botelho Claudia, Bernardo Stella M, Wormley Floyd, Lee Samuel A
a Division of Infectious Diseases ; University of New Mexico Health Science Center ; Albuquerque , NM USA.
Virulence. 2014;5(8):810-8. doi: 10.4161/21505594.2014.956648. Epub 2014 Oct 31.
We have previously demonstrated that the C. albicans pre-vacuolar protein sorting gene VPS4 is required for extracellular secretion of the secreted aspartyl proteases Sap2p and Saps4-6p. Furthermore, the vps4Δ null mutant has been shown to be markedly hypovirulent in a murine tail vein model of disseminated candidiasis. In these experiments, we sought to further define the role of the pre-vacuolar secretion pathway mediated by the pre-vacuolar sorting gene VPS4 in the pathogenesis of epithelial and mucosal infection using a broad range of virulence models. The C. albicans vps4Δ mutant demonstrates reduced tolerance of cell wall stresses compared to its isogenic, complemented control strain. In an in vitro oral epithelial model (OEM) of tissue invasion, the vps4Δ mutant caused reduced tissue damage compared to controls. Further, the vps4Δ mutant was defective in macrophage killing in vitro, and was attenuated in virulence in an in vivo Caenorhabditis elegans model representative of intestinal epithelial infection. In contrast, the vps4Δ mutant caused a similar degree of tissue damage in an in vitro uroepithelial model of Candida infection compared with controls. Furthermore, in an in vivo murine model of vaginal candidiasis there was no reduction in fungal colony burden and no differences in vaginal histopathology compared to wild-type and complemented controls. These results suggest that VPS4 contributes to several key aspects of oral epithelial but not uroepithelial infection, and in contrast to systemic infection, plays no major role in the pathogenesis of Candida vaginitis. By using a wide range of virulence models, we demonstrate that C. albicans VPS4 contributes to virulence according to the specific tissue that is infected. Thus, in order to gain a full understanding of C. albicans virulence in relation to a particular gene or pathway of interest, a selected range of infection models may need to be utilized.
我们之前已经证明,白色念珠菌液泡前体蛋白分选基因VPS4是分泌天冬氨酸蛋白酶Sap2p和Saps4 - 6p细胞外分泌所必需的。此外,vps4Δ缺失突变体在播散性念珠菌病的小鼠尾静脉模型中已被证明毒力明显减弱。在这些实验中,我们试图使用广泛的毒力模型,进一步确定由液泡前体分选基因VPS4介导的液泡前体分泌途径在上皮和黏膜感染发病机制中的作用。与同基因的互补对照菌株相比,白色念珠菌vps4Δ突变体对细胞壁应激的耐受性降低。在组织侵袭的体外口腔上皮模型(OEM)中,与对照相比,vps4Δ突变体导致的组织损伤减少。此外,vps4Δ突变体在体外巨噬细胞杀伤方面存在缺陷,并且在代表肠道上皮感染的体内秀丽隐杆线虫模型中毒力减弱。相比之下,在念珠菌感染的体外尿路上皮模型中,vps4Δ突变体与对照相比造成的组织损伤程度相似。此外,在阴道念珠菌病的体内小鼠模型中,与野生型和互补对照相比,真菌菌落负荷没有降低,阴道组织病理学也没有差异。这些结果表明,VPS4对口腔上皮感染的几个关键方面有贡献,但对尿路上皮感染没有贡献,并且与全身感染不同,在念珠菌性阴道炎的发病机制中不起主要作用。通过使用广泛的毒力模型,我们证明白色念珠菌VPS4根据所感染的特定组织对毒力有贡献。因此,为了全面了解白色念珠菌与特定感兴趣基因或途径相关的毒力,可能需要使用一系列选定的感染模型。