ALTERED MONOCYTE GENE EXPRESSION AND EXPANSION OF CD14+CD16+ SUBSET IN IgA NEPHROPATHY PATIENTS

The basic defect of IgA nephropathy (IgAN) lies within peripheral blood mononuclear cells rather than local kidney abnormalities. Previously we showed an altered gene expression in monocytes compared to B and T cells isolated from IgAN patients (Kidney Int, 2010), thus our aim here was to study this subset more closely at genome-wide level. total of 35 IgAN patients and 35 healthy blood donors (HBD) were included in this study. Illumina microarray technology was used to evaluate global differences in gene expression between monocytes isolated from IgAN patients and HBD. Bioinformatic analysis was performed with GenomeStudio and Genespring software. The connectivity between genes was evaluated using Ingenuity Pathway Analysis. Aberrantly expressed genes and pathways were then validated on an independent set of patients with RT-PCR western blot and flow cytometric analysis.

IgA肾病（IgA nephropathy, IgAN）的核心病理缺陷存在于外周血单个核细胞，而非肾脏局部的异常改变。此前本团队的研究显示，相较于从IgAN患者体内分离的B细胞与T细胞，其单核细胞的基因表达存在异常（Kidney Int, 2010），因此本研究旨在全基因组层面更深入地探究这一细胞亚群。本研究共纳入35例IgAN患者与35例健康献血者（healthy blood donors, HBD）。采用Illumina微阵列技术，对比分析IgAN患者与健康献血者分离得到的单核细胞的全基因组基因表达差异。使用GenomeStudio与Genespring软件开展生物信息学分析，并通过Ingenuity Pathway Analysis评估基因间的调控关联。随后，通过独立患者队列，采用RT-PCR、蛋白质印迹（western blot）与流式细胞术对异常表达的基因及通路进行验证。