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

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HIV Reservoirs

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2407))

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.

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Acknowledgments

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.

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Authors and Affiliations

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

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

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

    Jacob D. Estes

Authors
  1. Kathleen Busman-Sahay
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  2. Michael D. Nekorchuk
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  3. Carly Elizabeth Starke
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  4. Chi Ngai Chan
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  5. Jacob D. Estes
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Corresponding author

Correspondence to Jacob D. Estes .

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Editors and Affiliations

  1. School of Medicine and Human Immuno-Virology (H.I.V.) Group, San Raffaele University and Scientific Institute, Milano, Italy

    Guido Poli

  2. Viral Pathogenesis and Biosafety Group, San Raffaele Scientific Institute, Milano, Italy

    Elisa Vicenzi

  3. Department of Molecular and Comparative Pathobiology, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Fabio Romerio

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Busman-Sahay, K., Nekorchuk, M.D., Starke, C.E., Chan, C.N., Estes, J.D. (2022). In Situ Multiplexing to Identify, Quantify, and Phenotype the HIV-1/SIV Reservoir Within Lymphoid Tissue. In: Poli, G., Vicenzi, E., Romerio, F. (eds) HIV Reservoirs. Methods in Molecular Biology, vol 2407. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1871-4_19

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  • DOI: https://doi.org/10.1007/978-1-0716-1871-4_19

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1870-7

  • Online ISBN: 978-1-0716-1871-4

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