Cheng Shaoji, Clancy Cornelius J, Checkley Mary Ann, Handfield Martin, Hillman Jeffrey D, Progulske-Fox Ann, Lewin Alfred S, Fidel Paul L, Nguyen M Hong
Department of Medicine, University of Florida College of Medicine, Gainesville 32610, USA.
Mol Microbiol. 2003 Jun;48(5):1275-88. doi: 10.1046/j.1365-2958.2003.03521.x.
Candida albicans causes a wide spectrum of diseases, ranging from mucocutaneous infections like oral thrush to disseminated candidiasis. Screening for C. albicans genes expressed within infected hosts might advance understanding of candidal pathogenesis, but is impractical using existing techniques. In this study, we used an antibody-based strategy to identify C. albicans genes expressed during thrush. We adsorbed sera from HIV-infected patients with thrush against candidal cells grown in vitro and screened a C. albicans genomic expression library. We identified 10 genes encoding immunogenic antigens and used reverse transcription-polymerase chain reaction to verify that they were induced within thrush pseudomembranes recovered from a patient. The in vivo induced genes are involved in diverse functions, including regulation of yeast-hyphal morphogenesis, adhesion to host cells, nutrient uptake, phospholipid biosynthesis and amino acid catabolism. Four genes encode known virulence determinants (HWP1, CST20, CPP1 and RBF1). Another gene, LPD1, for which a role in candidal pathogenesis is unknown, encodes a protein homologous to a bacterial virulence determinant. Most importantly, disruption of CaNOT5, a newly identified gene, conferred defects in morphogenesis, decreased adherence to human buccal epithelial cells and attenuated mortality during murine disseminated candidiasis, proving that our strategy can identify genes encoding novel virulence determinants.
白色念珠菌可引发多种疾病,从口腔念珠菌病等黏膜皮肤感染到播散性念珠菌病。筛查感染宿主中表达的白色念珠菌基因可能会增进对念珠菌发病机制的理解,但使用现有技术并不实际。在本研究中,我们采用基于抗体的策略来鉴定鹅口疮期间表达的白色念珠菌基因。我们用来自患有鹅口疮的HIV感染患者的血清吸附体外培养的念珠菌细胞,并筛选白色念珠菌基因组表达文库。我们鉴定出10个编码免疫原性抗原的基因,并使用逆转录-聚合酶链反应来验证它们在从一名患者身上回收的鹅口疮假膜中被诱导表达。体内诱导的基因涉及多种功能,包括酵母-菌丝形态发生的调控、对宿主细胞的黏附、营养摄取、磷脂生物合成和氨基酸分解代谢。四个基因编码已知的毒力决定因素(HWP1、CST20、CPP1和RBF1)。另一个基因LPD1,其在念珠菌发病机制中的作用尚不清楚,编码一种与细菌毒力决定因素同源的蛋白质。最重要的是,新鉴定的基因CaNOT5的缺失导致形态发生缺陷、对人颊上皮细胞的黏附减少以及小鼠播散性念珠菌病期间死亡率降低,证明我们的策略可以鉴定编码新型毒力决定因素的基因。