Sequential staining of HIV gp140 to capture antigen-specific human B cells via flow cytometry

Protocols for efficient capture of antigen-specific B cells (ASBCs) are useful for understanding pathogen-specific B-cell responses during natural infection or vaccination. Fluorescently labeled tetramerized probes are classically used to capture ASBCs, but many occlude valuable epitopes available f...

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Autores principales: Townsley, Samantha M., Leggat, David J., Prabhakaran, Madhu, McDermott, Adrian B., Krebs, Shelly J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8390768/
https://www.ncbi.nlm.nih.gov/pubmed/34471908
http://dx.doi.org/10.1016/j.xpro.2021.100771
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author Townsley, Samantha M.
Leggat, David J.
Prabhakaran, Madhu
McDermott, Adrian B.
Krebs, Shelly J.
author_facet Townsley, Samantha M.
Leggat, David J.
Prabhakaran, Madhu
McDermott, Adrian B.
Krebs, Shelly J.
author_sort Townsley, Samantha M.
collection PubMed
description Protocols for efficient capture of antigen-specific B cells (ASBCs) are useful for understanding pathogen-specific B-cell responses during natural infection or vaccination. Fluorescently labeled tetramerized probes are classically used to capture ASBCs, but many occlude valuable epitopes available for B-cell receptor binding. Here, we describe a bead assay to confirm ASBC receptor accessibility on probes and a sequential staining process to capture HIV gp140-specific B cells from human peripheral blood mononuclear cells. For complete details on the use and execution of this protocol, please refer to Townsley et al. (2021).
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spelling pubmed-83907682021-08-31 Sequential staining of HIV gp140 to capture antigen-specific human B cells via flow cytometry Townsley, Samantha M. Leggat, David J. Prabhakaran, Madhu McDermott, Adrian B. Krebs, Shelly J. STAR Protoc Protocol Protocols for efficient capture of antigen-specific B cells (ASBCs) are useful for understanding pathogen-specific B-cell responses during natural infection or vaccination. Fluorescently labeled tetramerized probes are classically used to capture ASBCs, but many occlude valuable epitopes available for B-cell receptor binding. Here, we describe a bead assay to confirm ASBC receptor accessibility on probes and a sequential staining process to capture HIV gp140-specific B cells from human peripheral blood mononuclear cells. For complete details on the use and execution of this protocol, please refer to Townsley et al. (2021). Elsevier 2021-08-24 /pmc/articles/PMC8390768/ /pubmed/34471908 http://dx.doi.org/10.1016/j.xpro.2021.100771 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Townsley, Samantha M.
Leggat, David J.
Prabhakaran, Madhu
McDermott, Adrian B.
Krebs, Shelly J.
Sequential staining of HIV gp140 to capture antigen-specific human B cells via flow cytometry
title Sequential staining of HIV gp140 to capture antigen-specific human B cells via flow cytometry
title_full Sequential staining of HIV gp140 to capture antigen-specific human B cells via flow cytometry
title_fullStr Sequential staining of HIV gp140 to capture antigen-specific human B cells via flow cytometry
title_full_unstemmed Sequential staining of HIV gp140 to capture antigen-specific human B cells via flow cytometry
title_short Sequential staining of HIV gp140 to capture antigen-specific human B cells via flow cytometry
title_sort sequential staining of hiv gp140 to capture antigen-specific human b cells via flow cytometry
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8390768/
https://www.ncbi.nlm.nih.gov/pubmed/34471908
http://dx.doi.org/10.1016/j.xpro.2021.100771
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