Anti-tat Hutat2:Fc mediated protection against tat-induced neurotoxicity and HIV-1 replication in human monocyte-derived macrophages.
Anti-tat Hutat2:Fc mediated protection against tat-induced neurotoxicity and HIV-1 replication in human monocyte-derived macrophages.
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DOI：
10.1186/s12974-014-0195-2
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发表时间：
2014-11-22
影响因子：
9.3
通讯作者：
Lu Y
中科院分区：
文献类型：
--
作者：
Kang W;Marasco WA;Tong HI;Byron MM;Wu C;Shi Y;Sun S;Sun Y;Lu Y
HIV-1 Tat 对于 HIV 复制至关重要，也是众所周知的导致 HIV 相关神经认知障碍 (HAND) 的神经毒性因子。目前，一旦 HIV 感染确立，针对 HIV 逆转录酶或蛋白酶的联合抗逆转录病毒疗法无法阻止早期病毒蛋白的产生，尤其是 Tat。大脑中感染艾滋病毒的巨噬细胞和神经胶质细胞仍然将 Tat 释放到细胞外空间，在那里它可以发挥直接和间接的神经毒性。因此，大脑中稳定产生的抗Tat抗体将中和HIV-1 Tat，从而提供保护神经元的有效方法。我们构建了一种人源化抗 Tat Hutat2:Fc 融合蛋白，旨在拮抗 HIV-1 Tat，并通过基于 HIV 的慢病毒载体将该基因递送至细胞系和原代人单核细胞源性巨噬细胞 (hMDM) 中。体外评估了抗 Tat Hutat2:Fc 融合蛋白的功能以及慢病毒载体介导的基因转移的潜在副作用。我们的研究表明，基于 HIV-1 的慢病毒载体介导的基因转导导致抗 HIV-1 Tat Hutat2:Fc 在人类神经元和单核细胞系以及原代 hMDM 中高水平、稳定表达。 Hutat2:Fc 在细胞和上清液中均可检测到，并且在上清液中继续积累至高水平。 Hutat2:Fc 保护小鼠皮质神经元免受 HIV-1 Tat86 诱导的神经毒性。此外，分泌的 Hutat2:Fc 和转导的 hMDM 都会导致人类巨噬细胞中 HIV-1BaL 病毒复制的减少。此外，基于慢病毒载体的基因导入没有导致细胞形态和细胞活力的任何显着变化。尽管IL8、STAT1和IDO1基因的表达在转导的hMDM中上调，但这种基因表达的改变并不影响Hutat2:Fc的神经保护作用。我们的研究表明，慢病毒介导的基因转移可以有效地将 Hutat2:Fc 基因传递到原代 hMDM 中，并且不会导致 hMDM 免疫激活发生任何显着变化。 Hutat2:Fc 产生的神经保护和 HIV-1 抑制作用与全长抗 Tat 抗体相当。这项研究为未来的研究提供了基础和见解，即通过利用自然穿过血脑屏障的单核细胞/巨噬细胞进行基因传递，将 Hutat2:Fc 作为 HAND 的新型基因治疗方法的潜在用途。本文的在线版本 (doi:10.1186/s12974-014-0195-2) 包含补充材料，可供授权用户使用。
HIV-1 Tat is essential for HIV replication and is also a well-known neurotoxic factor causing HIV-associated neurocognitive disorder (HAND). Currently, combined antiretroviral therapy targeting HIV reverse transcriptase or protease cannot prevent the production of early viral proteins, especially Tat, once HIV infection has been established. HIV-infected macrophages and glial cells in the brain still release Tat into the extracellular space where it can exert direct and indirect neurotoxicity. Therefore, stable production of anti-Tat antibodies in the brain would neutralize HIV-1 Tat and thus provide an effective approach to protect neurons. We constructed a humanized anti-Tat Hutat2:Fc fusion protein with the goal of antagonizing HIV-1 Tat and delivered the gene into cell lines and primary human monocyte-derived macrophages (hMDM) by an HIV-based lentiviral vector. The function of the anti-Tat Hutat2:Fc fusion protein and the potential side effects of lentiviral vector-mediated gene transfer were evaluated in vitro. Our study demonstrated that HIV-1-based lentiviral vector-mediated gene transduction resulted in a high-level, stable expression of anti-HIV-1 Tat Hutat2:Fc in human neuronal and monocytic cell lines, as well as in primary hMDM. Hutat2:Fc was detectable in both cells and supernatants and continued to accumulate to high levels within the supernatant. Hutat2:Fc protected mouse cortical neurons against HIV-1 Tat86-induced neurotoxicity. In addition, both secreted Hutat2:Fc and transduced hMDM led to reducing HIV-1BaL viral replication in human macrophages. Moreover, lentiviral vector-based gene introduction did not result in any significant changes in cytomorphology and cell viability. Although the expression of IL8, STAT1, and IDO1 genes was up-regulated in transduced hMDM, such alternation in gene expression did not affect the neuroprotective effect of Hutat2:Fc. Our study demonstrated that lentivirus-mediated gene transfer could efficiently deliver the Hutat2:Fc gene into primary hMDM and does not lead to any significant changes in hMDM immune-activation. The neuroprotective and HIV-1 suppressive effects produced by Hutat2:Fc were comparable to that of a full-length anti-Tat antibody. This study provides the foundation and insights for future research on the potential use of Hutat2:Fc as a novel gene therapy approach for HAND through utilizing monocytes/macrophages, which naturally cross the blood-brain barrier, for gene delivery. The online version of this article (doi:10.1186/s12974-014-0195-2) contains supplementary material, which is available to authorized users.
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影响因子：
14.8
作者：
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