Mucosal Entry of HIV and Mucosal Immunity, Institut Cochin, Université de Paris, Paris, France.
INSERM U1016, Paris, France.
Methods Mol Biol. 2022;2407:189-204. doi: 10.1007/978-1-0716-1871-4_14.
HIV reservoirs in tissues are poorly understood and their establishment largely depends on the nature of tissues that interact with the virus. In this chapter, we will describe in vitro and ex vivo models of human urethral mucosal macrophages used in the investigation of the establishment and maintenance of tissue HIV reservoirs. In addition, we will describe how macrophage latent HIV infection was assessed in these models by reverting a nonproductive state of infection back into a productive state. Consequently, infectious particles are released to the macrophage extracellular milieu and detected by adapted viral outgrowth assays. Altogether, these approaches provide invaluable tools for the investigation on tissue-specific pathways that HIV-1 employs to reach host cells and form reservoirs in the genital mucosa. These models will contribute to the development of an efficient and targeted prophylaxis against HIV and of a HIV cure.
组织中的 HIV 储存库尚未被充分理解，其建立在很大程度上取决于与病毒相互作用的组织性质。在本章中，我们将描述用于研究组织 HIV 储存库建立和维持的人尿道黏膜巨噬细胞的体外和离体模型。此外，我们将描述如何通过将非生产性感染状态逆转回生产性状态来评估这些模型中的巨噬细胞潜伏 HIV 感染。结果，感染性颗粒被释放到巨噬细胞细胞外环境中，并通过适应性病毒扩增测定法检测到。总之，这些方法为研究 HIV-1 进入宿主细胞并在生殖道黏膜中形成储存库所利用的组织特异性途径提供了宝贵的工具。这些模型将有助于开发针对 HIV 的有效和靶向预防措施，并实现 HIV 的治愈。
