我国HIV-1肺储存库的准种遗传多样性和细胞嗜性研究

The lung is considered as one of the most important HIV-1 latent reservoirs. Most AIDS-associated opportunistic infections, such as tuberculosis, etc., can be infected by the respiratory tract (pulmonary). This project aims to establish an HIV-1-associated lung opportunistic infections cohort. To obtain quasispecies sequences of viral RNA and provirus DNA from peripheral blood mononuclear cells (PBMCs) and bronchoalveolar lavage fluid (BALF) from patients by single genomic amplification (SGA) and Next-generation sequencing (NGS) platforms. The subtype identification, genotypic tropism prediction and genetic diversity of peripheral blood and latent virus were carried out by the sequence analysis. In addition, the phenotypes of latent virus strains were further confirmed in vitro according to the cellular affinity experiments. This project will elucidate the relationship between HIV repository, cell tropism, and lung opportunistic infections among HIV-1 subtypes. It will provide an evidence for improving the antiviral therapy and finding new effective prevention and treatment methods for opportunistic infections of AIDS.

肺被认为是HIV-1感染患者非常重要的潜伏病毒储存库之一，大部分艾滋病机会性感染如肺结核等，可经由呼吸道（肺）感染。本项目通过建立HIV-1合并肺部机会性感染的研究队列，应用单模板基因组扩增技术（SGA）和二代测序（NGS）平台获得患者外周血和支气管肺泡灌洗液（BALF）的病毒RNA与单核细胞前病毒DNA准种序列，对外周血来源病毒和肺部潜伏病毒株序列进行亚型鉴定、基因型嗜性预测和遗传多样性分析；并根据细胞嗜性实验构建克隆进一步体外确认潜伏病毒株的表型嗜性。本项目将阐明我国HIV-1主要亚型的肺部HIV存储库特征和细胞嗜性转变与肺部机会性感染的关系，将为完善抗病毒治疗方案、寻找新的有效的艾滋病机会性感染防治方法提供理论依据。

肺被认为是HIV-1感染患者非常重要的潜伏病毒储存库之一，大部分艾滋病机会性感染如肺结核等，可经由呼吸道（肺）感染。本项目通过建立HIV-1合并肺部机会性感染的研究队列，应用单模板基因组扩增技术（SGA）获得患者外周血和支气管肺泡灌洗液（BALF）的HIV-1病毒RNA与单核细胞前病毒DNA准种序列，对外周血来源病毒和肺部潜伏病毒株序列进行亚型鉴定分析；通过对BALF和粪便来源的病原谱进行16S rRNA和18S rDNA测序分析，初步探索了HIV-1合并肺部机会性感染者体内病毒潜伏与肺部和肠道微生物多样性的关系，基本建成HIV测序分析技术以及准种进化分析平台。本项目初步阐述患者肺部细菌和真菌的多样性与肺部HIV-1病毒载量之间存在一定关系，肺部HIV-1病毒载量越低，肺部微生物群落越丰富；而抗病毒治疗对出现肺部机会性感染的患者的肠道菌群特征影响不大。本项目发现肺部机会性感染的发生发展主要与肺部HIV-1水平有关，为完善有效的艾滋病机会性感染防治方法提供理论依据。