Comparative Expression Profile of miRNA and mRNA in Primary Peripheral Blood Mononuclear Cells Infected with Human Immunodeficiency Virus (HIV-1) [mRNA]

Host cells respond to exogenous infectious agents such as viruses, including HIV-1. Studies have evaluated the changes associated with virus infection at the transcriptional and translational levels of the cellular genes involved in specific pathways. While this approach is useful, in our view it provides only a partial view of genome-wide changes. Recently, technological advances in the expression profiling at the microRNA (miRNA) and mRNA levels have made it possible to evaluate the changes in the components of multiple pathways. To understand the role of miRNA and its interplay with host cellular gene expression (mRNA) during HIV-1 infection, we performed a comparative global miRNA and mRNA microarray using human PBMCs infected with HIV-1. The PBMCs were derived from multiple donors and were infected with virus generated from the molecular clone pNL4-3. The results showed that HIV-1 infection led to altered regulation of 21 miRNAs and 444 mRNA more than 2-fold, with a statistical significance of p<0.05. Furthermore, the differentially regulated miRNA and mRNA were shown to be associated with host cellular pathways involved in cell cycle/proliferation, apoptosis, T-cell signaling, and immune activation. We also observed a number of inverse correlations of miRNA and mRNA expression in infected PBMCs, further confirming the interrelationship between miRNA and mRNA regulation during HIV-1 infection. These results for the first time provide evidence that the miRNA profile could be an early indicator of host cellular dysfunction induced by HIV-1. Total RNA isolated from PBMC subjected to 7 days of HIV-1 infection compared to uninfected control PBMC

宿主细胞可对包括人类免疫缺陷病毒1型（HIV-1）在内的外源性感染因子产生应答。已有研究针对参与特定通路的细胞基因的转录与翻译水平，分析了病毒感染引发的表达变化。尽管该研究思路具备一定应用价值，但我们认为其仅能呈现基因组范围变化的局部视图。近年来，微小RNA（miRNA）与信使RNA（mRNA）水平的表达谱分析技术取得长足进步，使得同时解析多条细胞通路组分的表达变化成为可能。为阐明HIV-1感染过程中miRNA的作用及其与宿主细胞基因表达（mRNA）的相互调控关系，我们以感染HIV-1的人外周血单个核细胞（PBMC）为实验材料，开展了全基因组范围的miRNA与mRNA芯片联合分析。本次实验所用PBMC取自多名健康供体，所使用的病毒由分子克隆pNL4-3包装制备。研究结果显示，HIV-1感染可使21种miRNA与444条mRNA的表达水平发生超过2倍的显著变化（P<0.05）。进一步分析表明，这些差异表达的miRNA与mRNA富集于细胞周期/增殖、细胞凋亡、T细胞信号通路以及免疫激活等宿主细胞通路中。此外，我们在感染HIV-1的PBMC中观测到多组miRNA与mRNA表达呈负相关关系，进一步验证了HIV-1感染过程中miRNA与mRNA调控的相互关联性。本研究首次证实，miRNA表达谱可作为HIV-1诱导宿主细胞功能异常的早期检测标志物。本研究提取的总RNA来自感染HIV-1 7天的PBMC，并以未感染的PBMC作为对照。