In Situ Multiplexing to Identify, Quantify, and Phenotype the HIV-1/SIV Reservoir Within Lymphoid Tissue

Kathleen Busman-Sahay, Michael D. Nekorchuk, Carly Elizabeth Starke, Chi Ngai Chan, Jacob D. Estes

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Modern combination antiretroviral therapy (ART) regimens provide abiding viral suppression for most individuals infected with human immunodeficiency virus (HIV). However, the persistence of viral reservoirs ensures that eradication of HIV-1 (i.e., cure) or sustained ART-free remission (i.e., functional cure) remains elusive, necessitating continual, strict ART adherence and contributing to HIV-1-related comorbidities. Eradication of these viral reservoirs, which persist primarily within lymphoid tissue, will require a deeper understanding of the cellular neighborhoods in which latent and active HIV-1-infected cells reside. By pairing highly sensitive in situ hybridization (ISH) with an exceptionally flexible immunofluorescence (IF) approach, we describe a simple, yet highly adaptable multiplex protocol for investigating the quantity, distribution, and characteristics of HIV-1 viral reservoirs.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages277-290
Number of pages14
DOIs
StatePublished - 2022
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume2407
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Fluorescent multiplex
  • HIV
  • Human immunodeficiency virus
  • IF
  • Immunofluorescence
  • In situ hybridization
  • ISH
  • Lymphoid tissue
  • Reservoir

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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