Inhibition of human immunodeficiency virus type 1 replication in vitro by a novel combination of anti-Tat single-chain intrabodies and NF-kappa B antagonists
Inhibition of human immunodeficiency virus type 1 replication in vitro by a novel combination of anti-Tat single-chain intrabodies and NF-kappa B antagonists
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DOI：
10.1128/jvi.71.9.6486-6494.1997
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发表时间：
1997-09-01
影响因子：
5.4
通讯作者：
Marasco, WA
中科院分区：
文献类型：
--
作者：
Mhashilkar, AM;Biswas, DK;Marasco, WA
人类免疫缺陷病毒1型（HIV-1）达特是一种早期调节蛋白，对病毒基因表达和复制至关重要，通过与反式激活反应元件（TAR）结合反式激活HIV-1长末端重复序列（LTR），并与其他细胞因子沿着增加病毒转录起始和延伸，达特还通过TAR非依赖性机制超激活HIV-1启动子，包括肿瘤坏死因子α诱导的和蛋白激酶C（PKC）依赖的NF-κ B活化，达特和NF-κ B的抑制剂协同下调这种Tat介导的LTR超活化。在这项研究中，使用两种PRC（NF-κ B）抑制剂，戊茶碱（PTX）和Go-6976，以及稳定表达的抗Tat单链胞内抗体（sFv intrabody）的组合药理学和遗传学策略，以获得对HIV-1 LTR驱动的基因表达和HIV-1复制的协同抑制。用PTX和Go-6976处理细胞导致HIV-1 LTR驱动的基因表达和HIV-1复制的协同抑制。此外，抗Tat sFv胞内抗体和两种NF-κ B抑制剂的组合使用使病毒保持潜伏状态长达45天，与单独使用NF-κ B抑制剂或单独使用抗Tar sFv胞内抗体相比，联合治疗导致对HIV-1复制的更持久抑制。总之，这些结果表明，在未来的临床基因治疗试验中，像这里报道的那样，结合药理学和遗传学策略可能会改善转导细胞的存活率并延长临床获益。
Human immunodeficiency virus type 1 (HIV-1) Tat, an early regulatory protein that is critical for viral gene expression and replication, transactivates the HIV-1 long terminal repeat (LTR) via its binding to the transactivation response element (TAR) and, along with other cellular factors, increases viral transcription initiation and elongation, Tat also superactivates the HIV-1 promoter through a TAR-independent mechanism, including tumor necrosis factor alpha-induced and protein kinase C (PKC)-dependent activation of NF-kappa B, and inhibitors of Tat and NF-kappa B cooperatively down-regulate this Tat-mediated LTR superactivation. In this study, a combined pharmacologic and genetic strategy using two PRC (NF-kappa B) inhibitors, pentoxifylline (PTX) and Go-6976, and a stably expressed anti-Tat single-chain intracellular antibody (sFv intrabody) was employed to obtain cooperative inhibition of both HIV-1 LTR-driven gene expression and HIV-1 replication. Treatment of cells with PTX and Go-6976 resulted in cooperative inhibition of both HIV-1 LTR-driven gene expression and HIV-1 replication, In addition, the combined use of anti-Tat sFv intrabodies and the two NF-kappa B inhibitors retained the virus in the latent state for as long as 45 days, The combined treatment resulted in more durable inhibition of HIV-1 replication than was seen with the NF-kappa B inhibitors alone or the anti-Tar sFv intrabodies alone. Together, these results suggest that in future clinical gene therapy trials, a combined pharmacologic and genetic strategy like the one reported here may improve the survival of transduced cells and prolong clinical benefit.