A highly efficient short hairpin RNA potently down-regulates CCR5 expression in systemic lymphoid organs in the hu-BLT mouse model
A highly efficient short hairpin RNA potently down-regulates CCR5 expression in systemic lymphoid organs in the hu-BLT mouse model
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DOI：
10.1182/blood-2009-04-215855
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发表时间：
2010-02-25
期刊：
影响因子：
20.3
通讯作者：
An, Dong Sung
中科院分区：
文献类型：
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作者：
Shimizu, Saki;Hong, Patrick;An, Dong Sung
抑制HIV-1辅助受体CCR 5的表达对于控制患者的HIV-1感染具有很大的希望。在这里，我们报告稳定敲低人CCR 5的短发夹RNA（shRNA）在人源化骨髓/肝脏/胸腺（BLT）小鼠模型。我们通过慢病毒载体转导将针对CCR 5的有效shRNA导入人胎肝来源的CD 34（+）造血祖细胞/干细胞（HPSC）。我们将载体转导的HPSC用Matrigel和胸腺段固化在小鼠肾包膜下。随后将载体转导的自体CD 34（+）细胞注射到辐照小鼠中，旨在建立全身重建。CCR 5表达下调的人T细胞和单核细胞/巨噬细胞在全身淋巴组织，包括肠道相关淋巴组织，HIV-1复制的主要网站。通过多克隆T细胞受体V β家族发育和幼稚/记忆T细胞分化评估，shRNA介导的CCR 5敲低对T细胞发育没有明显的不利影响。在二次移植的小鼠中CCR 5敲低表明通过shRNA转导的HPSC的长期造血重建的潜力。CCR 5嗜性HIV-1感染在小鼠来源的人脾细胞中被有效地抑制。这些结果表明，慢病毒载体将shRNA递送到人HPSC中可以稳定地下调体内全身淋巴器官中的CCR 5。（血。2010;115：1534-1544）
Inhibiting the expression of the HIV-1 coreceptor CCR5 holds great promise for controlling HIV-1 infection in patients. Here we report stable knockdown of human CCR5 by a short hairpin RNA(shRNA) in a humanized bone marrow/liver/thymus (BLT) mouse model. We delivered a potent shRNA against CCR5 into human fetal liver-derived CD34(+) hematopoietic progenitor/stem cells (HPSCs) by lentiviral vector transduction. We transplanted vector-transduced HPSCs solidified with Matrigel and a thymus segment under the mouse kidney capsule. Vector-transduced autologous CD34(+) cells were subsequently injected in the irradiated mouse, intended to create systemic reconstitution. CCR5 expression was down-regulated in human T cells and monocytes/macrophages in systemic lymphoid tissues, including gut-associated lymphoid tissue, the major site of HIV-1 replication. The shRNA-mediated CCR5 knockdown had no apparent adverse effects on T-cell development as assessed by polyclonal T-cell receptor V beta family development and naive/memory T-cell differentiation. CCR5 knockdown in the secondary transplanted mice suggested the potential of long-term hematopoietic reconstitution by the shRNA-transduced HPSCs. CCR5 tropic HIV-1 infection was effectively inhibited in mouse-derived human splenocytes ex vivo. These results demonstrate that lentiviral vector delivery of shRNA into human HPSCs could stably down-regulate CCR5 in systemic lymphoid organs in vivo. (Blood. 2010;115:1534-1544)