Highly Sensitive Analysis of Cervical Mucosal HIV-1 Infection Using Reporter Viruses Expressing Secreted Nanoluciferase

Methods Mol Biol. 2024:2807:299-323. doi: 10.1007/978-1-0716-3862-0_21.

Abstract

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.

Keywords: Cervical mucosa; Cervix; Ex vivo model; HIV-1; Nanoluciferase; Reporter virus; Transmission.

MeSH terms

  • Cervix Uteri* / metabolism
  • Cervix Uteri* / virology
  • Female
  • Genes, Reporter
  • HIV Infections* / virology
  • HIV-1* / genetics
  • HIV-1* / physiology
  • Humans
  • Luciferases / genetics
  • Luciferases / metabolism
  • Mucous Membrane / metabolism
  • Mucous Membrane / virology
  • Virus Replication