Barker Katherine S, Liu Teresa, Rogers P David
Department of Pharmacy, College of Pharmacy, University of Tennessee Health Science Center, Memphis, Tennessee, USA.
J Infect Dis. 2005 Sep 1;192(5):901-12. doi: 10.1086/432487. Epub 2005 Jul 20.
The host's first line of defense against bloodstream infection with Candida albicans involves the recognition and clearance of the fungus by neutrophils and monocytes/macrophages. The purpose of the present study was to examine changes in the monocytic cell gene-expression profile in response to C. albicans stimulation.
RNA was isolated from THP-1 cells 3 h after coculture with live C. albicans SC5314 cells. After hybridization to microarrays, genes differentially expressed by at least 2.0-fold were included in the final data set.
As expected, TNFA, IL8, CD83, MIP1A, and MIP1B were among the genes up-regulated. This was confirmed by real-time reverse-transcriptase polymerase chain reaction (RT-PCR), fluorescence-activated cell sorting analysis, and enzyme-linked immunosorbent assay. Furthermore, RGS1, RGS2, RGS16, DSCR1, GROB, EGR3, FLT4, and TNFAIP6 were also up-regulated in response to C. albicans, whereas CCR2 and NCF2 were among the genes down-regulated in response to C. albicans. Differential expression of selected genes was confirmed at several time points by real-time RT-PCR.
This study defines the gene expression profile of an early response of human mononuclear cells to C. albicans and identifies genes not previously known to be responsive to this pathogen.
宿主抵御白色念珠菌血流感染的第一道防线涉及中性粒细胞和单核细胞/巨噬细胞对真菌的识别和清除。本研究的目的是检测单核细胞基因表达谱对白色念珠菌刺激的反应变化。
将THP-1细胞与活的白色念珠菌SC5314细胞共培养3小时后分离RNA。与微阵列杂交后，最终数据集中纳入至少2.0倍差异表达的基因。
正如预期，TNFA、IL8、CD83、MIP1A和MIP1B是上调基因之一。这通过实时逆转录聚合酶链反应（RT-PCR）、荧光激活细胞分选分析和酶联免疫吸附测定得到证实。此外，RGS1、RGS2、RGS16、DSCR1、GROB、EGR3、FLT4和TNFAIP6也对白色念珠菌有上调反应，而CCR2和NCF2是对白色念珠菌下调的基因。通过实时RT-PCR在几个时间点证实了所选基因的差异表达。
本研究确定了人类单核细胞对白色念珠菌早期反应的基因表达谱，并鉴定出以前未知对该病原体有反应的基因。
