Glycosylphosphatidylinositol-Anchored Anti-HIV scFv Efficiently Protects CD4 T Cells from HIV-1 Infection and Deletion in hu-PBL Mice
Glycosylphosphatidylinositol-Anchored Anti-HIV scFv Efficiently Protects CD4 T Cells from HIV-1 Infection and Deletion in hu-PBL Mice
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DOI：
10.1128/jvi.01389-16
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发表时间：
2016-11
影响因子：
5.4
通讯作者：
Chaobaihui Ye;Weiming Wang;L. Cheng;Guangming Li;M. Wen;Qi Wang;Qing Zhang;Danxiu Li;P. Zhou;L. Su
中科院分区：
文献类型：
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作者：
Chaobaihui Ye;Weiming Wang;L. Cheng;Guangming Li;M. Wen;Qi Wang;Qing Zhang;Danxiu Li;P. Zhou;L. Su
摘要尽管高效抗逆转录病毒治疗（HAART）成功地抑制了病毒和恢复了CD 4 T细胞，但由于治疗期间HIV-1储库的持续存在，HIV-1仍然无法治愈。1例急性髓性白血病患者接受了来自纯合子CCR 5 Δ32供体的异基因造血干细胞移植，在HAART停止后9年未检测到病毒血症。这个案例激发了一个HIV-1治疗研究领域，该领域专注于在允许细胞中设计HIV-1抗性。在此，我们采用糖基磷脂酰肌醇（GPI）-scFv X5方法来赋予人原代CD 4 T细胞对HIV-1的抗性。我们发现表达GPI-scFv X5的原代CD 4 T细胞对CCR 5（R5）-、CXCR 4（X4）-和双嗜性HIV-1具有抗性，并且与离体对照细胞相比具有存活优势。在hu-PBL小鼠研究中，在HIV-I感染后在外周血和淋巴组织中选择GPI-scFv X5转导的CD 4 T细胞。最后，GPI-scFv X5转导的CD 4 T细胞在与HIV感染的细胞共输注后，与GPI-scFv AB 65转导的CD 4 T细胞的小鼠相比，hu-PBL小鼠中相对于CD 4细胞显示出显著降低的病毒载量和病毒RNA拷贝数。我们得出结论，GPI-scFv X5修饰的CD 4 T细胞可潜在地用作针对R5和X4嗜性HIV-1感染的遗传干预。重要性阻断HIV-1进入是最有前途的治疗方法之一。通过T细胞中的核酸酶对HIV-1辅助受体CCR 5的遗传破坏正在进行2项临床试验，并导致患者的病毒血症减少。然而，使用CXCR 4辅助受体的病毒的出现是应用单辅助受体破坏的疗法的关注点。在这里，我们报告了用GPI-scFv X5工程化的HIV-1-容许性CD 4 T细胞对R5-、X4-或双嗜性病毒感染具有体外抗性。在使用hu-PBL小鼠的临床前研究中，我们表明CD 4 T细胞受到保护，并且在HIV-1感染的动物中选择了GPI-scFv X5转导的细胞。此外，我们表明GPI-scFv X5转导的CD 4 T细胞对体内病毒复制产生负面影响。我们得出的结论是，GPI-scFv X5修饰的CD 4 T细胞有可能用作针对R5和X4嗜性HIV-1感染的遗传干预措施。
ABSTRACT Despite success in viral inhibition and CD4 T cell recovery by highly active antiretroviral treatment (HAART), HIV-1 is still not curable due to the persistence of the HIV-1 reservoir during treatment. One patient with acute myeloid leukemia who received allogeneic hematopoietic stem cell transplantation from a homozygous CCR5 Δ32 donor has had no detectable viremia for 9 years after HAART cessation. This case has inspired a field of HIV-1 cure research focusing on engineering HIV-1 resistance in permissive cells. Here, we employed a glycosylphosphatidylinositol (GPI)-scFv X5 approach to confer resistance of human primary CD4 T cells to HIV-1. We showed that primary CD4 T cells expressing GPI-scFv X5 were resistant to CCR5 (R5)-, CXCR4 (X4)-, and dual-tropic HIV-1 and had a survival advantage compared to control cells ex vivo. In a hu-PBL mouse study, GPI-scFv X5-transduced CD4 T cells were selected in peripheral blood and lymphoid tissues upon HIV-1 infection. Finally, GPI-scFv X5-transduced CD4 T cells, after being cotransfused with HIV-infected cells, showed significantly reduced viral loads and viral RNA copy numbers relative to CD4 cells in hu-PBL mice compared to mice with GPI-scFv AB65-transduced CD4 T cells. We conclude that GPI-scFv X5-modified CD4 T cells could potentially be used as a genetic intervention against both R5- and X4-tropic HIV-1 infections. IMPORTANCE Blocking of HIV-1 entry is one of most promising approaches for therapy. Genetic disruption of the HIV-1 coreceptor CCR5 by nucleases in T cells is under 2 clinical trials and leads to reduced viremia in patients. However, the emergence of viruses using the CXCR4 coreceptor is a concern for therapies applying single-coreceptor disruption. Here, we report that HIV-1-permissive CD4 T cells engineered with GPI-scFv X5 are resistant to R5-, X4-, or dual-tropic virus infection ex vivo. In a preclinical study using hu-PBL mice, we show that CD4 T cells were protected and that GPI-scFv X5-transduced cells were selected in HIV-1-infected animals. Moreover, we show that GPI-scFv X5-transduced CD4 T cells exerted a negative effect on virus replication in vivo. We conclude that GPI-scFv X5-modified CD4 T cells could potentially be used as a genetic intervention against both R5- and X4-tropic HIV-1 infections.