宿主细胞调控HIV-1复制和潜伏的分子机制

发布时间：2019-07-09 18:50

【摘要】：第一部分:宿主因子Naf1调控HIV-1复制人类免疫缺陷病毒(Human Immunodeficiency Virus,HIV-1)属于逆转录病毒,其感染细胞、完成复制周期主要包括以下几个环节:病毒通过受体和辅佐受体结合宿主靶细胞(主要是CD4+T细胞)、脱壳、逆转录合成双链c DNA进入细胞核然后整合到宿主基因组上、病毒基因的转录和剪切、病毒m RNA至细胞质的转运、病毒蛋白的翻译和病毒颗粒的包装、出芽和成熟等。深入研究HIV-1劫持和利用宿主系统的机制将帮助我们寻找和鉴定针对宿主蛋白的抗病毒药物和基因治疗方法。我们利用基因组学筛选了多种潜在调控HIV-1复制的宿主蛋白,其中Naf1(HIV-1 Nef-associated factor 1)是一种依赖于CRM1(Chromosome region maintenance 1)可在细胞核/质间穿梭的宿主因子,在炎症反应中调控多种受体介导的信号通路。定位在细胞质中的Naf1可以通过结合A20蛋白来抑制NF-κB的活化,这种由Naf1引起的NF-κB失活与宿主多种自身免疫疾病的发病有关。然而Naf1对HIV-1感染和复制的影响并不清楚。我们的研究发现Naf1可促进HIV-1未剪切gag m RNA的出核;Naf1这一功能依赖于与另外一个宿主因子CRM1的相互作用,并且与Naf1在细胞核/质间穿梭特性相关。本研究揭示了宿主因子Naf1调控HIV-1复制机制,为抗病毒药物设计和基因治疗提供了潜在的宿主靶点。第二部分树突状细胞激活潜伏HIV-1的机制研究尽管高效抗逆转录病毒治疗(Highly active antiretroviral therapy,HAART)能够显著降低HIV-1病毒感染者血浆病毒载量,延长病人生命,但潜伏/持续性感染的HIV-1不能被药物根除,这是当前HIV/AIDS治疗的难点之一。目前对HIV-1潜伏感染建立、维持和再激活机制还不太清楚。树突状细胞(Dendritic cells,DC)作为重要的抗原递呈细胞在抗病原体免疫反应中能激活T细胞,并在HIV-1感染过程中发挥双刃剑作用。在本研究中,我们探究了DCs在诱导潜伏HIV-1激活中的作用,发现DCs通过产生抗潜伏细胞因子TNF-α激活CD4+T细胞中潜伏HIV-1的复制。与HIV-1潜伏感染CD4+T细胞共培养能促进DCs成熟和TNF-α释放,这一过程依赖CD40-CD40L信号通路;此外,我们的研究结果证明,AIDS相关病原体刺激也能诱导DCs成熟,促进DCs表面CD40表达,从而增强DCs激活潜伏HIV-1的能力。深入理解DCs介导的潜伏HIV-1的激活机制将有助于我们提出新的抗病毒潜伏治疗方案,开发有效清除HIV-1病毒库的治疗策略。
文内图片：

图片说明：Rev介导HIV-1mRNA的出核
[Abstract]:The first part: host factor Naf1 modulates HIV-1 replication of human immunodeficiency virus (Human Immunodeficiency Virus,HIV-1) belongs to retrovirus, and its replication cycle mainly includes the following links: virus binds to host target cells (mainly CD4 T cells) through receptor and auxiliary receptor, shelled, reverse transcriptional synthesis of double stranded c DNA enters the nucleus and integrates into the host genome. The transcription and shearing of viral genes, the transport of virus m RNA to cytoplasm, the translation of viral proteins and the packaging of virus particles, budding and maturation, etc. Further study on the mechanism of HIV-1 hijacking and utilization of host system will help us to find and identify antiviral drugs and gene therapy methods for host proteins. We used genomics to screen a variety of host proteins that potentially regulate HIV-1 replication. Among them, Naf1 (HIV-1 Nef-associated factor 1) is a host factor that depends on CRM1 (Chromosome region maintenance 1), which can shuttle between nucleus and cytoplasm and regulate multiple receptor-mediated signaling pathways in inflammatory response. Naf1 located in cytoplasm can inhibit the activation of NF- kappa B by binding A20 protein. This inactivation of NF- kappa B caused by Naf1 is related to the pathogenesis of a variety of host autoimmune diseases. However, the effect of Naf1 on HIV-1 infection and replication is not clear. Our study found that Naf1 can promote the nucleation of uncut gag m RNA in HIV-1, and the function of Naf1 depends on the interaction with another host factor CRM1 and is related to the shuttling characteristics of Naf1 between nucleus and cytoplasm. This study revealed the mechanism of host factor Naf1 regulating HIV-1 replication, which provided a potential host target for antiviral drug design and gene therapy. The second part is the mechanism of dendritic cells activating latent HIV-1 although highly effective antiretrovirus therapy for (Highly active antiretroviral therapy,HAART can significantly reduce the plasma viral load and prolong the life of patients infected with HIV-1 virus, but the latent / persistent infection HIV-1 can not be eliminated by drugs, which is one of the difficulties in the treatment of HIV/AIDS. At present, the mechanism of establishing, maintaining and reactivating HIV-1 latent infection is not very clear. Dendritic cells (Dendritic cells,DC), as important antigen presenting cells, can activate T cells in the immune response of antifungal chlamydia and play a double-edged role in the process of HIV-1 infection. In this study, we investigated the role of DCs in inducing latent HIV-1 activation, and found that DCs activated the replication of latent HIV-1 in CD4 T cells by producing anti-latent cytokine TNF- 伪. Co-culture with HIV-1 latent infected CD4 T cells can promote DCs maturation and TNF- 伪 release, which depends on CD40-CD40L signaling pathway. In addition, our results show that AIDS related pathogen stimulation can also induce DCs maturation and promote CD40 expression on DCs surface, thus enhancing the ability of DCs to activate latent HIV-1. Understanding the activation mechanism of DCs-mediated latent HIV-1 will help us to propose a new antiviral latent treatment scheme and develop an effective treatment strategy for clearing HIV-1 virus library.
【学位授予单位】：苏州大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R512.91

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