在淋巴组织中鉴定、定量和表型分析 HIV-1/SIV 储存库的原位多重分析。

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

机构信息

Vaccine & Gene Therapy Institute, Oregon Health & Science University, Beaverton, OR, USA.

Division of Pathobiology & Immunology, Oregon National Primate Research Center, Oregon Health & Science University, Portland, OR, USA.

出版信息

Methods Mol Biol. 2022;2407:277-290. doi: 10.1007/978-1-0716-1871-4_19.

DOI:10.1007/978-1-0716-1871-4_19

PMID:34985671

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.

摘要

现代联合抗逆转录病毒疗法（ART）方案为大多数感染人类免疫缺陷病毒（HIV）的个体提供持久的病毒抑制。然而，病毒储存库的持续存在确保了 HIV-1 的根除（即治愈）或持续无 ART 缓解（即功能性治愈）仍然难以实现，这需要持续、严格的 ART 依从性，并导致与 HIV-1 相关的合并症。消除这些主要存在于淋巴组织中的病毒储存库，需要更深入地了解潜伏和活跃的 HIV-1 感染细胞所在的细胞邻里关系。通过将高灵敏度原位杂交（ISH）与极其灵活的免疫荧光（IF）方法相结合，我们描述了一种简单但高度适应的多重方案，用于研究 HIV-1 病毒储存库的数量、分布和特征。