Battivelli Emilie, Verdin Eric
Gladstone Institute of Virology and Immunology, Gladstone Institutes, San Francisco, CA, USA.
Department of Medicine, University of California San Francisco, San Francisco, CA, USA.
Bio Protoc. 2018 Oct 20;8(20). doi: 10.21769/bioprotoc.3050.
While able to suppress HIV replication in HIV infected individuals, combination antiretroviral therapy (ART) fails to eliminate viral latent reservoir, which consists in integrated transcriptional silenced HIV provirus. So far, identification of latently-infected cells has relied on activating cells to induce expression of HIV proteins which can then be detected. Unfortunately, this activation significantly changed the cell phenotype. We developed a novel HIV reporter, named HIV, that allows the purification of latently-infected cells in absence of reactivation. Indeed, latent cells can be identified by expression of the EF1a-driven mKO2 and lack of expression of the LTR-driven csGFP. This protocol can be used to study the effectiveness of LRAs (Latency Reversal Agents) in reactivating latent HIV in primary cells.
虽然联合抗逆转录病毒疗法（ART）能够抑制HIV感染者体内的HIV复制，但无法清除由整合的转录沉默HIV前病毒组成的病毒潜伏库。到目前为止，潜伏感染细胞的鉴定依赖于激活细胞以诱导HIV蛋白表达，然后对其进行检测。不幸的是，这种激活显著改变了细胞表型。我们开发了一种名为HIV的新型HIV报告基因，它能够在不进行再激活的情况下纯化潜伏感染细胞。实际上，潜伏细胞可以通过EF1a驱动的mKO2的表达以及LTR驱动的csGFP的不表达来鉴定。该方案可用于研究潜伏逆转剂（LRAs）在激活原代细胞中潜伏HIV方面的有效性。
