Revealing HIV-1 persistence in myeloid cell reservoirs

摘要 在有效的抗逆转录病毒疗法面前，评估髓系细胞是否支持HIV-1的持久性是一个重要的 技术挑战。因此，到目前为止，还没有直接证据表明髓系细胞在病毒中扮演任何角色。 在有效的艺术面前坚持。因此，对髓系细胞储存库的研究正在落入 默默无闻。巨噬细胞的感染只能由能够使用低水平的HIV-1变异体来启动 细胞表面的CD_4分子。因此，如果一个功能正常的髓系储存库有助于病毒在 有效的ART，我们可以预测在分析治疗中断(ATI)后反弹的病毒血症，将 含有对CD4有很高亲和力的病毒变体。我们已经开发出一种方法，可以识别 ATI后反跳性病毒血症的低频率巨噬细胞嗜性变异体。通过单基因组 扩增(SGA)，我们从ATI患者的血浆中克隆了大量的病毒包膜。 当这些包膜被用来构建重组分子克隆时，它们赋予了融合的能力 与初级巨噬细胞结合，并在其中复制。我们认为，这些结果提供了确凿的证据 在抑制ART的感染者中存在髓系细胞库，此外， 当ART被中断时，这个储存库有助于病毒反弹。 我们假设髓系细胞库是稳定的，允许病毒在抑制下长期存在。 ART和燃料在治疗中断时病毒反弹。为了进一步研究这一假设，我们建议： 1：通过回弹纵向抽样评估嗜巨噬细胞病毒的频率 急性心肌梗死后病毒血症。 2：评估长期缓解者的髓系细胞库的持久性 骨髓移植后的间隔时间(波士顿患者)以及复发性病毒血症的采样 来自经历了连续ATI的个体。 3：评估嗜巨噬细胞病毒是否起源于中枢神经系统，以及它们是否不太适合 T嗜性病毒。 这项提议将结合临床、病毒学和分子研究来揭示髓系细胞的存在。 对水库个体的抑制艺术，并阐明这一水库的动态。关键人物 从这项研究中获得的信息将提供是否有理由扩大开发努力 以髓系细胞库为靶点并加速病毒治愈的战略。

Abstract Assessing whether myeloid cells support HIV-1 persistence in the face of effective ART represents a significant technical challenge. As a result, there is as yet, no direct evidence that myeloid cells play any role in viral persistence in the face of effective ART. As a consequence, research on myeloid cell reservoirs is falling into obscurity. Infection of macrophages can only be initiated by HIV-1 variants that have the ability to use low levels of CD4 on the cell surface. Therefore, if a functional myeloid reservoir contributes to viral persistence under effective ART, we would predict that viremia that rebounds following analytic treatment interruption (ATI), would contain viral variants that have a high affinity for CD4. We have developed an approach that allows identification of low frequency macrophage-tropic variants in rebounding viremia post ATI. Through single genome amplification (SGA), we cloned a large number of viral envelopes from plasma of individuals who underwent ATI. When these envelopes were used to construct recombinant molecular clones, they conferred the ability to fuse with, and replicate within primary macrophages. We believe that these results provide definitive evidence for the existence of a myeloid cell reservoir in infected individuals on suppressive ART and furthermore, that this reservoir contributes to viral rebound when ART is interrupted. We hypothesize that the myeloid cell reservoir is stable, permits long term viral persistence under suppressive ART and fuels viral rebound upon treatment interruption. To pursue this hypothesis, we propose to: 1: Assess the frequency of macrophage-tropic viruses through longitudinal sampling of rebounding viremia post-ATI. 2: Assess the persistent nature of the myeloid cell reservoir from individuals with prolonged remission intervals following bone marrow transplant (Boston patients) as well as sampling of rebounding viremia from individuals who underwent sequential ATI. 3: Evaluate whether macrophage-tropic viruses have a CNS origin and whether they are less fit relative to T tropic viruses. This proposal will integrate clinical, virologic and molecular studies to reveal the existence of a myeloid cell reservoir in individuals on suppressive ART and to shed light on the dynamics of this reservoir. The critical information gained from this study will provide whether there is rationale for an expanded effort to develop strategies to target the myeloid cell reservoir and to accelerate the path to a viral cure.