Exposing virus-induced long noncoding RNA in plasmacytoid dendritic cells

摘要 浆细胞样树突状细胞（pDC）是先天性免疫应答的重要组成部分，有助于 通过产生大量的抗病毒I型干扰素和调节 适应性反应我们的实验室已经阐明了人类pDC激活中的许多关键途径， 在一些实施方案中，所述方法包括使所述细胞成熟，特别是关于ssRNA病毒如HIV-1和流感病毒。我们展示了 这是pDC首次通过Toll样受体（TLR）7， 需要CD 4依赖性内化到内体区室的过程，其中下游IFN 信号通过干扰素调节因子7（IRF）激活。尽管触发相同的TLR（TLR 7），HIV-1 和流感对pDC的影响不同。HIV-1诱导大量I型IFN，但不能使pDC完全成熟 进入抗原呈递细胞或APC（“IFN”表型对“APC”表型），而流感诱导两者 表型值得注意的是，我们还表明，HIV-2激活pDC成为APC，类似于流感。HIV-2是一种 自然减毒形式的艾滋病毒，这往往导致控制病毒和延迟疾病的发展。 我们现在已经确定了一组长的非编码RNA（lncRNA），可以调节人类的常规免疫反应。 DC（cDC）和pDC功能，包括它们的IFN和成熟/APC表型。我们还描述了 人pDC中的IFN-诱导型lncRNA组以及可能潜在影响pDC的独特lncRNA pDC响应病毒感应而发挥作用。本应用程序的总体目标是调查病毒的作用 在pDC调节中诱导的lncRNA，鉴于病毒可以诱导lncRNA，lncRNA是重要的调节剂 基因及其编码蛋白的表达和功能，以及lncRNAS影响免疫细胞 行为，包括树突状细胞。我们假设ssRNA病毒（HIV-1，流感病毒，HIV-2）可以 通过其调节性lnc-RNA的差异诱导影响pDC功能。具体目标是： (1)应用RNA测序鉴定人pDC中独特的病毒诱导的lncRNA;（2）功能上定义 在pDC中调节IFN应答的lncRNA，和（3）机械地区分在pDC中分离IFN应答的lncRNA。 pDC对HIV-1和HIV-2的应答。 我们目前对lncRNA在感染的先天免疫细胞中的身份和功能的了解是非常有限的。 有限，特别是对人类先天免疫细胞。鉴定病毒诱导的lncRNA可能导致 发现新的细胞途径，以帮助开发治疗工具， 根除HIV-1感染。

Abstract Plasmacytoid dendritic cells (pDC) are an important component of the innate immune response, contributing to the immune control of viral infections by producing large amounts of anti-viral type I interferon and modulating adaptive responses. Our laboratory has elucidated many of the key pathways in human pDC activation and maturation, especially with regard to ssRNA viruses such as HIV-1 and influenza virus. We demonstrated for the first time that pDC sense HIV-1 associated single stranded RNA through Toll-like receptor (TLR) 7, a process that requires CD4-dependent internalization into endosomal compartments where downstream IFN signaling is activated via interferon regulatory factor 7 (IRF). Despite triggering the same TLR (TLR7), HIV-1 and influenza differentially impact pDC. HIV-1 induces large amounts of type I IFN but fails to fully mature pDC into antigen presenting cells or APC (“IFN” phenotype vs “APC” phenotype), whereas influenza induces both phenotypes. Notably, we also show that HIV-2 activates pDC to become APC, similar to influenza. HIV-2 is a naturally attenuated form of HIV, which results often in control of the virus and delayed disease development. We have now identified a set of long noncoding RNAs (lncRNAs) that can modulate both human conventional DC (cDC) and pDC function, including their IFN and maturation/APC phenotypes. We have also described sets of IFN-inducible lncRNAs in human pDC as well as pDC-unique lncRNAs that could potentially impact pDC function in response to virus sensing. The overall goal of this application is to investigate the role of virus induced-lncRNA in pDC regulation, given that viruses can induce lncRNAs, lncRNAs are important regulators of the expression and function of genes and their encoded proteins, and that lncRNAS affect immune cell behavior, including that of dendritic cells. We hypothesize that ssRNA viruses (HIV-1, influenza, HIV-2) can impact pDC function through their differential induction of regulatory lnc-RNAs. The specific aims are to: (1) Apply RNA sequencing to identify unique virus-induced lncRNA in human pDC; (2) Functionally define lncRNA that modulate the IFN response in pDC, and (3) Mechanistically distinguish lncRNA that segregate pDC responses to HIV-1 and HIV-2. Our current knowledge about the identity and function of lncRNAs in infected innate immune cells is very limited, especially for human innate immune cells. Identifying virus inducible lncRNAs may lead to the discovery of novel cellular pathways to aid the development of therapeutic tools for the treatment and eradication of HIV-1 infection.