TY - CHAP
T1 - In Situ Multiplexing to Identify, Quantify, and Phenotype the HIV-1/SIV Reservoir Within Lymphoid Tissue
AU - Busman-Sahay, Kathleen
AU - Nekorchuk, Michael D.
AU - Starke, Carly Elizabeth
AU - Chan, Chi Ngai
AU - Estes, Jacob D.
N1 - Funding Information:
We acknowledge support from the National Institutes of Health grant AI143411-01A1 (J.D.E) and the Oregon National Primate Research Center grant award P51OD011092. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Publisher Copyright:
© 2022, Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2022
Y1 - 2022
N2 - 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.
AB - 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.
KW - Fluorescent multiplex
KW - HIV
KW - Human immunodeficiency virus
KW - IF
KW - ISH
KW - Immunofluorescence
KW - In situ hybridization
KW - Lymphoid tissue
KW - Reservoir
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U2 - 10.1007/978-1-0716-1871-4_19
DO - 10.1007/978-1-0716-1871-4_19
M3 - Chapter
C2 - 34985671
AN - SCOPUS:85122780249
T3 - Methods in Molecular Biology
SP - 277
EP - 290
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -