Baxter Amy E, Niessl Julia, Morou Antigoni, Kaufmann Daniel E
Research Centre of the Centre Hospitalier de l'Université de Montréal (CRCHUM), and Université de Montréal, Montreal, QC, Canada.
Center for HIV/AIDS Vaccine Immunology and Immunogen Discovery (CHAVI-ID), La Jolla, CA, USA.
AIDS Res Ther. 2017 Sep 12;14(1):40. doi: 10.1186/s12981-017-0171-x.
Despite the tremendous success of anti-retroviral therapy (ART) no current treatment can eradicate latent HIV reservoirs from HIV-infected individuals or generate, effective HIV-specific immunity. Technological limitations have hampered the identification and characterization of both HIV-infected cells and HIV-specific responses in clinical samples directly ex vivo. RNA-flow cytometric fluorescence in situ hybridisation (RNA Flow-FISH) is a powerful technique, which enables detection of mRNAs in conjunction with proteins at a single-cell level. Here, we describe how we are using this technology to address some of the major questions remaining in the HIV field in the era of ART. We discuss how CD4 T cell responses to HIV antigens, both following vaccination and HIV infection, can be characterized by measurement of cytokine mRNAs. We describe how our development of a dual HIV mRNA/protein assay (HIV assay) enables high sensitivity detection of very rare HIV-infected cells and aids investigations into the translation-competent latent reservoir in the context of HIV cure.
尽管抗逆转录病毒疗法（ART）取得了巨大成功，但目前尚无治疗方法能够从HIV感染者体内根除潜伏的HIV储存库，也无法产生有效的HIV特异性免疫。技术限制阻碍了对直接取自临床样本的HIV感染细胞和HIV特异性反应进行体外鉴定和表征。RNA流式细胞荧光原位杂交技术（RNA Flow-FISH）是一项强大的技术，能够在单细胞水平上结合蛋白质检测mRNA。在此，我们描述了如何利用这项技术来解决ART时代HIV领域中仍然存在的一些主要问题。我们讨论了如何通过测量细胞因子mRNA来表征接种疫苗和感染HIV后CD4 T细胞对HIV抗原的反应。我们描述了我们开发的双重HIV mRNA/蛋白质检测方法（HIV检测方法）如何实现对极其罕见的HIV感染细胞的高灵敏度检测，并有助于在HIV治愈的背景下对具有翻译能力的潜伏储存库进行研究。
