RNA interference-mediated silencing of the YPS3 gene of Histoplasma capsulatum reveals virulence defects
RNA interference-mediated silencing of the YPS3 gene of Histoplasma capsulatum reveals virulence defects
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DOI：
10.1128/iai.00304-07
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发表时间：
2007-06-01
影响因子：
3.1
通讯作者：
Woods, Jon P.
中科院分区：
文献类型：
--
作者：
Bohse, Megan L.;Woods, Jon P.
荚膜组织胞浆菌YPS 3基因编码一种蛋白质，该蛋白质位于荚膜组织胞浆菌细胞壁的表面。包囊并释放到培养基中。这种蛋白质仅在致病性酵母感染阶段产生，并且在H.不同毒力水平的荚膜菌株。在这项研究中，我们沉默的YPS 3转录使用干扰RNA策略，并检查了沉默的突变体在体外和感染过程中的表型差异。突变体在37 ° C下在限定培养基中体外培养期间没有显示出生长缺陷，并且似乎在RAW 264.7鼠巨噬细胞样细胞系中具有正常的毒力。然而，在C57 BL/6小鼠感染模型中，突变体导致真菌负荷显著降低，特别是在肝脏和脾脏的外周吞噬细胞丰富的组织中。器官定植的这种缺陷在感染后3天内很明显;然而，它似乎在以后的时间点加重。
The YPS3 gene of Histoplasma capsulatum encodes a protein that is both surface localized in the cell wall of H. capsulatum and released into the culture medium. This protein is produced only during the pathogenic yeast phase of infection and is also expressed differentially in H. capsulatum strains of different virulence levels. In this study, we silenced the YPS3 transcript by using an interfering-RNA strategy and examined the silenced mutants for phenotypic differences in vitro and during infection. The mutants showed no growth defect during in vitro culture in a defined medium at 37 degrees C and appeared to have normal virulence in a RAW 264.7 murine macrophage-like cell line. In a C57BL/6 mouse model of infection, however, the mutants caused significantly decreased fungal burdens, particularly in the peripheral phagocyte-rich tissues of livers and spleens. This defect in organ colonization was evident within 3 days of infection; however, it appeared to be exacerbated at later time points.