1. Sequential staining of HIV gp140 to capture antigen-specific human B cells via flow cytometry.
- Author
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Townsley SM, Leggat DJ, Prabhakaran M, McDermott AB, and Krebs SJ
- Subjects
- Humans, Leukocytes, Mononuclear immunology, Molecular Probes, Receptors, Antigen, B-Cell metabolism, B-Lymphocytes immunology, Flow Cytometry methods, Staining and Labeling methods, env Gene Products, Human Immunodeficiency Virus chemistry
- Abstract
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)., Competing Interests: The authors declare no competing interests., (© 2021 The Author(s).)
- Published
- 2021
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