Determine the Transcriptional Activity of Non-integrated HIV-1 DNA

描述（由申请方提供）：人类免疫缺陷病毒（HIV）感染导致CD4 T细胞耗竭，并导致免疫抑制状态。病毒复制需要其基因组稳定整合到人类染色体中，然而HIV感染的自然过程导致大量未整合的病毒DNA在体内积累。尽管在体外的非整合的病毒DNA的转录活性的明确证明，在了解这个早期过程的调节和它在HIV生命周期中的直接作用方面取得了很小的进展。因此，未整合的病毒DNA在HIV感染中的致病潜力尚未得到很好的理解。本申请的长期目标是了解调节整合前转录的分子机制以及该早期过程在病毒发病机制中的贡献。该提议的具体目的是确定调节整合前转录的病毒DNA模板，以及HIV达特参与整合前转录的起始。实现这些特定目标的实验方法是使用HIV整合酶阴性病毒和我们最近构建的表征整合前转录的新型指示细胞。预计从这些研究中获得的数据将有助于深入了解非整合的病毒DNA能够产生病毒因子，增强病毒复制并促进病毒发病机制的情况。这项研究将有助于了解非整合的HIV DNA在疾病进展中的潜在致病作用。此外，在某些情况下，非整合的病毒DNA可以作为疾病进展的临床标志物。

DESCRIPTION (provided by applicant): Infection by the Human Immunodeficiency Virus (HIV) causes depletion of CD4 T cells and leads to a state of immuno-suppression. The viral replication requires stable integration of its genome into the human chromosome, yet the natural process of HIV infection leads to the accumulation of a large quantity of unintegrated viral DNA in the body. In spite of clear demonstration of transcriptional activity from non-integrated viral DNA in vitro, little progress has been made in understanding regulation of this early process and its direct role in the HIV life cycle. As such, pathogenic potential of non- integrated viral DNA in HIV infection is not well understood. The long-term objectives of this application are to understand the molecular mechanisms that regulate pre- integration transcription and the contribution of this early process in viral pathogenesis. The specific aims of this proposal are to determine the viral DNA template regulating pre-integration transcription, and the involvement of HIV Tat in the initiation of pre- integration transcription. The experiment methods to achieve these specific aims are to use an HIV integrase negative virus and a novel indicator cell that we have recently constructed to characterize pre-integration transcription. It is expected that data acquired from these studies will make help to gain insight into circumstances where non-integrated viral DNA is capable of producing viral factors, enhancing viral replication and contributing to viral pathogenesis. The proposed research will help to understand potential pathogenic role of non-integrated HIV DNA in disease progression. Additionally, non-integrated viral DNA could serve as clinical markers for disease progression in certain situation.