Heinrich Pette Institute - Leibniz Institute for Experimental Virology, Hamburg, Germany.
PLoS Pathog. 2013;9(9):e1003587. doi: 10.1371/journal.ppat.1003587. Epub 2013 Sep 26.
Stable integration of HIV proviral DNA into host cell chromosomes, a hallmark and essential feature of the retroviral life cycle, establishes the infection permanently. Current antiretroviral combination drug therapy cannot cure HIV infection. However, expressing an engineered HIV-1 long terminal repeat (LTR) site-specific recombinase (Tre), shown to excise integrated proviral DNA in vitro, may provide a novel and highly promising antiviral strategy. We report here the conditional expression of Tre-recombinase from an advanced lentiviral self-inactivation (SIN) vector in HIV-infected cells. We demonstrate faithful transgene expression, resulting in accurate provirus excision in the absence of cytopathic effects. Moreover, pronounced Tre-mediated antiviral effects are demonstrated in vivo, particularly in humanized Rag2⁻/⁻γc⁻/⁻ mice engrafted with either Tre-transduced primary CD4⁺ T cells, or Tre-transduced CD34⁺ hematopoietic stem and progenitor cells (HSC). Taken together, our data support the use of Tre-recombinase in novel therapy strategies aiming to provide a cure for HIV.
HIV 前病毒 DNA 稳定整合到宿主细胞染色体中，这是逆转录病毒生命周期的标志和基本特征，使感染永久化。目前的抗逆转录病毒联合药物治疗不能治愈 HIV 感染。然而，表达一种经过工程改造的 HIV-1 长末端重复（LTR）位点特异性重组酶（Tre），已证明可在体外切除整合的前病毒 DNA，可能提供一种新颖且极具前景的抗病毒策略。我们在此报告在 HIV 感染的细胞中，从先进的慢病毒自我失活（SIN）载体中条件表达 Tre 重组酶。我们证明了忠实的转基因表达，在没有细胞病变效应的情况下导致准确的前病毒切除。此外，在体内证明了明显的 Tre 介导的抗病毒作用，特别是在用人 Rag2⁻/⁻γc⁻/⁻小鼠移植 Tre 转导的原代 CD4⁺ T 细胞或 Tre 转导的 CD34⁺造血干细胞和祖细胞（HSC）的情况下。总之，我们的数据支持在旨在为 HIV 提供治愈方法的新型治疗策略中使用 Tre 重组酶。
