Mycobacterium bovis BCG cell wall-specific differentially expressed genes identified by differential display and cDNA subtraction in human macrophages
Mycobacterium bovis BCG cell wall-specific differentially expressed genes identified by differential display and cDNA subtraction in human macrophages
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DOI：
10.1128/iai.72.2.937-948.2004
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发表时间：
2004-02-01
影响因子：
3.1
通讯作者：
Seya, T
中科院分区：
文献类型：
--
作者：
Begum, NA;Ishii, K;Seya, T
我们分析了单核细胞对高度纯化的牛分枝杆菌BCG细胞壁部分的基因表达谱，这是一种临床批准的佐剂，称为BCG细胞壁骨架（BCG- cws）。它由霉菌酸、阿拉伯半乳聚糖和肽聚糖组成，并赋予toll样受体2 （TLR2）和tlr4依赖性信号，诱导单核细胞分化为抗原呈递细胞（APCs）。在这里，我们报告了bcg - cws刺激与非刺激单核细胞的差异基因表达分析。BCG-CWS大量诱导受TLR信号调控的基因。与脂多糖（LPS）刺激的细胞相比，bcg - cws刺激的细胞具有显著的基因调控特征。(i)编码TREM-1和TREM-2可溶性形式（最近发现的炎症信号放大受体）的剪接mrna受BCG-CWS调节，导致其差异表达。（ii）锌铁转运蛋白（ZIP）样家族蛋白HKE-1和LIV-1基因被BCG-CWS单独诱导。（iii） BCG-CWS诱导的是白介素-23 (IL-23)，而非IL-12p70，而干扰素诱导基因仅由LPS诱导。通过Northern和逆转录- pcr分析，我们确认了30多个BCG-CWS调控基因的差异表达，并将其与LPS和其他已知TLR配体的表达进行了比较。一组基因对LPS反应迅速且反应时间短，但对BCG-CWS反应时间长。对鉴定出的新蛋白或假设蛋白的结构分析显示，其中一些可能是信号介质或转录调节因子的潜在候选者。因此，BCG-CWS可能以一种不同于LPS的方式深刻调节APC反应，这可能导致其辅助活性治疗潜力的优势。
We have analyzed the gene expression profile of monocytes in response to a highly purified cell wall fraction of Mycobacterium bovis BCG, a clinically approved adjuvant known as BCG cell wall skeleton (BCG-CWS). It is composed of mycolic acid, arabinogalactan, and peptidoglycan and confers Toll-like receptor 2 (TLR2)- and TLR4-dependent signaling that induces monocytes to differentiate into antigen-presenting cells (APCs). Here we report differential gene expression analysis with BCG-CWS-stimulated versus nonstimulated monocytes. BCG-CWS exerted massive induction of genes regulated by TLR signaling. Marked gene regulatory characteristics in BCG-CWS-stimulated cells compared to lipopolysaccharide (LPS)-stimulated cells follow. (i) Spliced mRNAs encoding soluble forms of TREM-1 and TREM-2 (recently discovered inflammatory-signal-amplifying receptors) were regulated by BCG-CWS, resulting in their differential expression. (ii) The genes for zinc-iron transporter protein (ZIP)-like family proteins HKE-1 and LIV-1 were induced exclusively by BCG-CWS. (iii) Interleukin-23 (IL-23), rather than IL-12p70, was induced by BCG-CWS, while interferon-inducible genes were induced only by LPS. By Northern and reverse transcription-PCR analyses, we confirmed the differential expression of more than 30 BCG-CWS regulatory genes, and their expression was compared with that of LPS and other known TLR ligands. A battery of genes responded rapidly and for a short time to LPS but for a long time to BCG-CWS. Structural analysis of the identified novel or hypothetical proteins revealed that some are potential candidates as signaling mediators or transcriptional regulators. Hence, BCG-CWS may profoundly modulate APC responses in a way distinct from that of LPS, leading to possible advantages for its adjuvant-active therapeutic potential.