Enhanced gene transfer to mouse dendritic cells using adenovioral vectors coated with a novel adapter molecule
Enhanced gene transfer to mouse dendritic cells using adenovioral vectors coated with a novel adapter molecule
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DOI：
10.1016/j.ymthe.2004.02.006
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发表时间：
2004-05-01
影响因子：
12.4
通讯作者：
Blackwell, JL
中科院分区：
文献类型：
--
作者：
Pereboev, AV;Nagle, JM;Blackwell, JL
腺病毒(Ad)介导的树突状细胞(DC)转导效率低下，这是因为缺乏主要的Ad受体CAR。靶向CD40的重组腺病毒感染DC后，基因转移增加。目前的报告描述了CD40靶向方法的进一步发展，该方法使用一种适配器分子，将ADS的纤维连接到小鼠DC上的CD40。接头分子CFm40L由CAR通过三聚化基序与小鼠CD40配体融合而成。建立了高水平分泌CFm40L的稳定细胞系。用CFm40L靶向的Ad向小鼠骨髓来源的DC(MBMDC)转移基因的效率比非靶向Ad高4个数量级以上。超过70%的mBMDC实现了基因转移，而不是使用非靶向广告进行不可检测的转导。除了显著增强基因转移外，CFm40L靶向的Ads还诱导表型成熟和上调IL-12的表达。最重要的是，CFm40L靶向的Adds在体内诱导了针对模型抗原的特异性免疫反应。本研究的结果表明，利用非固有的转导途径，如CD40途径，可以显著增强Ad介导的DC基因转移，这可能在癌症和传染病的基因疫苗中有重要的应用。
Adenovirus (Ad)-mediated transduction of dendritic cells (DC) is inefficient because of the lack of the primary Ad receptor, CAR. DC infection with Ad targeted to the CD40 results in increased gene transfer. The current report describes further development of the CD40-targeting approach using an adapter molecule that bridges the fiber of the Ads to CD40 on mouse DC. The adapter molecule, CFm40L, consists of CAR fused to mouse CD40 ligand via a trimerization motif. A stable cell line that secretes CFm40L at high levels was generated. Gene transfer to mouse bone marrow-derived DC (mBMDC) using CFm40L-targeted Ad was over 4 orders of magnitude more efficient than that for the untargeted Ads. Gene transfer was achieved to over 70% of the mBMDC compared to undetectable transduction using untargeted Ads. In addition to dramatically enhanced gene transfer, the CFm40L-targeted Ads induced phenotypical maturation and upregulated IL-12 expression. Most importantly, the CFm40L-targeted Ads elicited specific immune response against a model antigen in vivo. The results of this study demonstrate that Ad-mediated gene transfer to DC can be significantly enhanced using nonnative transduction pathways, such the CD40 pathway, which may have important applications in genetic vaccination for cancer and infectious diseases.