Department of Biological Science, College of Natural Sciences, Ajou University, Suwon, Korea.
Haemophilia. 2012 May;18(3):e247-53. doi: 10.1111/j.1365-2516.2011.02702.x. Epub 2011 Dec 19.
Inhibitor development is the most significant complication in the therapy of haemophilia A (HA) patients. In spite of many studies, not much is known regarding the mechanism underlying inhibitor development. To understand the mechanism, we analysed profiles of differentially expressed genes (DEGs) between inhibitor and non-inhibitor HA via a microarray technique. Twenty unrelated Korean HAs were studied: 11 were non-inhibitor and nine were HA with inhibitor (≥5 BU mL(-1)). Microarray analysis was conducted using a Human Ref-8 expression Beadchip system (Illumina) and the data were analysed using Beadstudio software. We identified 545 DEGs in inhibitor HA as compared with the non-inhibitor patients; 384 genes were up-regulated and 161 genes were down-regulated. Among them, 75 genes whose expressions were altered by at least two-fold (>+2 or <-2) were selected and classified via the PANTHER classification method. The expressions of signal transduction and immunity-related genes differed significantly in the two groups. For validation of the DEGs, semi-quantitative RT-PCR (semi-qRT-PCR) was conducted with the six selected DEGs. The results corresponded to the microarray data, with the exception of one gene. We also examined the expression of the genes associated with the antigen presentation process via real-time PCR. The average levels of IL10, CTLA4 and TNFα slightly reduced, whereas that of IFNγ increased in the inhibitor HA group. We are currently unable to explain whether this phenomenon is a function of the inhibitor-inducing factor or is an epiphenomenon of antibody production. Nevertheless, our results provide a possible explanation for inhibitor development.
抑制剂的产生是治疗 A 型血友病（HA）患者的最主要并发症。尽管有很多研究，但对于抑制剂产生的机制仍知之甚少。为了理解这一机制，我们通过微阵列技术分析了抑制剂和非抑制剂 HA 之间差异表达基因（DEGs）的谱。研究了 20 名无关的韩国 HA 患者：11 名是非抑制剂，9 名为有抑制剂（≥5 BU mL(-1)）的 HA。使用 Human Ref-8 Expression Beadchip 系统（Illumina）进行微阵列分析，并用 Beadstudio 软件分析数据。与非抑制剂患者相比，我们在抑制剂 HA 中发现了 545 个 DEGs；其中 384 个基因上调，161 个基因下调。其中，选择了 75 个表达至少改变两倍（>+2 或 <-2）的基因，并通过 PANTHER 分类方法进行分类。两组间信号转导和免疫相关基因的表达有明显差异。为了验证 DEGs，我们用 6 个选定的 DEGs 进行了半定量 RT-PCR（semi-qRT-PCR）。结果与微阵列数据相符，除了一个基因。我们还通过实时 PCR 检查了与抗原呈递过程相关基因的表达。在抑制剂 HA 组中，IL10、CTLA4 和 TNFα 的平均水平略有降低，而 IFNγ 的水平升高。我们目前还无法解释这种现象是抑制剂诱导因子的功能，还是抗体产生的一种伴随现象。然而，我们的结果为抑制剂的产生提供了一种可能的解释。
