Baculovirus Transduction of Mesenchymal Stem Cells Triggers the Toll-Like Receptor 3 Pathway
Baculovirus Transduction of Mesenchymal Stem Cells Triggers the Toll-Like Receptor 3 Pathway
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DOI：
10.1128/jvi.01250-09
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发表时间：
2009-10-15
影响因子：
5.4
通讯作者：
Hu, Yu-Chen
中科院分区：
文献类型：
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作者：
Chen, Guan-Yu;Shiah, Hsiao-Chiao;Hu, Yu-Chen
人类间充质干细胞（hMSCs）可以用病毒载体进行遗传修饰，并有望成为再生医学的细胞来源，但hMSCs对病毒载体转导的反应仍然知之甚少，安全性问题尚未得到解决。在这里，我们探讨了hMSCs对一种新兴的DNA病毒载体，杆状病毒（BV）的反应，并发现BV转导扰乱了与5个信号通路相关的816个基因的转录。令人惊讶的是，Toll样受体-3（TLR 3），一种通常识别双链RNA的受体，通过BV转导明显上调，如微阵列，PCR阵列，流式细胞术和共聚焦显微镜所证实的。细胞因子阵列数据显示BV转导触发了白细胞介素-6（IL-6）和IL-8的稳健分泌，但未触发其他炎性细胞因子和β干扰素（IFN-β）的稳健分泌。BV转导激活了信号分子（例如，Toll/白细胞介素-1受体结构域的衔接子诱导的IFN-β，NF-κ B，和IFN调节因子3）的TLR 3的下游，而沉默TLR 3基因的小干扰RNA相当大地消除细胞因子的表达和促进细胞迁移。这些数据首次证明，DNA病毒载体可以激活hMSC中的TLR 3途径，并导致与免疫细胞不同的细胞因子表达谱。这些发现强调了评估TLR 3信号级联是否在由其他DNA载体引起的免疫应答中发挥作用的重要性。例如，在一个实施例中，腺病毒）。尽管如此，BV转导几乎不干扰表面标志物表达，仅诱导短暂和轻度的细胞因子应答，从而缓解了使用BV进行hMSCs工程的安全性问题。
Human mesenchymal stem cells (hMSCs) can be genetically modified with viral vectors and hold promise as a cell source for regenerative medicine, yet how hMSCs respond to viral vector transduction remains poorly understood, leaving the safety concerns unaddressed. Here, we explored the responses of hMSCs against an emerging DNA viral vector, baculovirus (BV), and discovered that BV transduction perturbed the transcription of 816 genes associated with five signaling pathways. Surprisingly, Toll-like receptor-3 (TLR3), a receptor that generally recognizes double-stranded RNA, was apparently upregulated by BV transduction, as confirmed by microarray, PCR array, flow cytometry, and confocal microscopy. Cytokine array data showed that BV transduction triggered robust secretion of interleukin-6 (IL-6) and IL-8 but not of other inflammatory cytokines and beta interferon (IFN-beta). BV transduction activated the signaling molecules (e.g., Toll/interleukin-1 receptor domain-containing adaptor-inducing IFN-beta, NF-kappa B, and IFN regulatory factor 3) downstream of TLR3, while silencing the TLR3 gene with small interfering RNA considerably abolished cytokine expression and promoted cell migration. These data demonstrate, for the first time, that a DNA viral vector can activate the TLR3 pathway in hMSCs and lead to a cytokine expression profile distinct from that in immune cells. These findings underscore the importance of evaluating whether the TLR3 signaling cascade plays roles in the immune response provoked by other DNA vectors (e. g., adenovirus). Nonetheless, BV transduction barely disturbed surface marker expression and induced only transient and mild cytokine responses, thereby easing the safety concerns of using BV for hMSCs engineering.