Division of Hematology/Oncology, Department of Medicine, Heersink School of Medicine, University of Alabama at Birmingham, Birmingham, AL, USA.
Birmingham VA Health Care System, Research Service, Birmingham, AL, USA.
Methods Mol Biol. 2024;2807:299-323. doi: 10.1007/978-1-0716-3862-0_21.
Ex vivo cervical tissue explant models offer a physiologically relevant approach for studying virus-host interactions that underlie mucosal HIV-1 transmission to women. However, the utility of cervical explant tissue (CET) models has been limited for both practical and technical reasons. These include assay variation, inadequate sensitivity for assessing HIV-1 infection and replication in tissue, and constraints imposed by the requirement for using multiple replica samples of CET to test each experimental variable and assay parameter. Here, we describe an experimental approach that employs secreted nanoluciferase (sNLuc) and current HIV-1 reporter virus technologies to overcome certain limitations of earlier ex vivo CET models. This method augments application of the CET model for investigating important questions involving mucosal HIV-1 transmission.
离体宫颈组织外植体模型为研究病毒-宿主相互作用提供了一种生理相关的方法,这些相互作用是导致 HIV-1 经黏膜向女性传播的基础。然而,由于实际和技术原因,宫颈外植体组织 (CET) 模型的应用受到了限制。这些原因包括检测变异性、评估组织中 HIV-1 感染和复制的敏感性不足,以及需要使用多个 CET 复制品样本来测试每个实验变量和检测参数所带来的限制。在这里,我们描述了一种实验方法,该方法采用分泌型纳米荧光素酶 (sNLuc) 和当前的 HIV-1 报告病毒技术,克服了早期离体 CET 模型的某些限制。这种方法增强了 CET 模型在研究涉及黏膜 HIV-1 传播的重要问题方面的应用。