Tetracycline-inducible interleukin-10 gene transfer mediated by an adeno-associated virus:: Application to experimental arthritis
Tetracycline-inducible interleukin-10 gene transfer mediated by an adeno-associated virus:: Application to experimental arthritis
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DOI：
10.1089/104303402320138961
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发表时间：
2002-07-01
影响因子：
4.2
通讯作者：
Jorgensen, C
中科院分区：
文献类型：
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作者：
Apparailly, F;Millet, V;Jorgensen, C
腺相关病毒（AAV）为类风湿关节炎（RA）的细胞因子基因转移提供了新的前景，因为它们是非致病性的，并允许在体内长期转基因表达。此外，四环素诱导型启动子的使用允许调节治疗性基因表达。本研究评估了表达病毒白细胞介素-10（vIL-10）的重组AAV载体在人类风湿性滑膜中的潜在长期基因调控以及在RA小鼠模型中的治疗效率。我们构建了一个重组腺相关病毒载体，其中vIL-10 cDNA的转录由TetON系统控制。用AAV-tetON-vIL 10转导人原代RA滑膜细胞赋予体外受控的vIL-10表达。肌内注射后，与对照组相比，在免疫和加强后用AAV-TetON-vIL 10载体加多西环素处理的DBA 1小鼠组中，胶原诱导的关节炎的发病率和严重程度在宏观、放射学和组织学水平上均显著降低。当共注射两个单独的AAV载体时，一个编码诱导型vIL-10，另一个编码转录激活因子，过量10倍的反式激活因子载体剂量允许在8周的时间内通过多西环素给药或停药有效控制vIL-10的分泌。我们的研究结果首次支持了AAV-tetON-vIL 10作为RA基因治疗的治疗工具的实用性。
The adeno-associated viruses (AAV) offer new perspectives for cytokine gene transfer in rheumatoid arthritis (RA) because they are nonpathogenic and allow long-term transgene expression in vivo. Moreover, the use of a tetracycline-inducible promoter allows regulation of therapeutic gene expression. This study assessed the potential long-term gene regulation of a recombinant AAV vector expressing viral interleukin-10 (vIL-10) in human rheumatoid synovium and the therapeutic efficiency in a mouse model of RA. We constructed a recombinant AAV vector in which the transcription of vIL-10 cDNA is controlled by the TetON system. Transduction of human primary RA synovial cells with AAV-tetON-vIL10 conferred in vitro controlled vIL-10 expression. After intramuscular injection, both incidence and severity of collagen-induced arthritis were significantly reduced at macroscopic, radiological, and histological levels in the group of DBA1 mice treated with AAV-TetON-vIL10 vector plus doxycycline after immunization and boosting compared to control groups. When coinjecting two separate AAV vectors, one encoding the inducible vIL-10 and the other the transcriptional activator, a 10 times excess of the transactivator vector dose allowed efficient control of vIL-10 secretion by doxycycline administration or withdrawal, over an 8-week period. Our results supported, for the first time, the utility of AAV-tetON-vIL10 as a therapeutic tool for gene therapy in RA.