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Updated: Dec 13, 2025

Detection and Enrichment of Rare Antigen-specific B Cells for Analysis of Phenotype and Function
Published on: February 16, 2017
OMIP-068: High-Dimensional Characterization of Global and Antigen-Specific B Cells in Chronic Infection
Katherine Cascino1, Mario Roederer2, Thomas Liechti2
1Viral Hepatitis Center, Department of Infectious Disease, Johns Hopkins University School of Medicine, Baltimore, Maryland, 21205, USA.
Abstract:
This 24-color flow cytometry panel focuses on characterizing antigen-specific B cells and precise delineation of B-cell subsets in chronic infections and is applicable to other chronic diseases such as autoimmunity. The panel was optimized for human cryopreserved peripheral blood mononuclear cells (PBMCs). Markers were chosen to extensively distinguish B-cell lineages (CD19, CD20, CD10, CD38, CD24, IgM, IgD, CD27, CD21, CD43, CD5). Inclusion of antigen-specific probes was of high priority in order to assess hepatitis B virus (HBV) antigen-specific B cells for our purposes. These probes can be readily exchanged for other pathogen-specific probes or additional markers for the panel to be tailored to desired research questions beyond HBV. In addition, we included a comprehensive and unique set of functional markers such as chemokine receptors (CXCR3, CXCR5), co-stimulatory molecule (CD86), Fc receptor (CD32), regulatory molecules (BTLA, CD39), and inhibitory markers associated with chronic infections (PD-1, FcRL5, CD11c, CD22) to enable in-depth analysis of global and antigen-specific B cells during chronic infection. © 2020 International Society for Advancement of Cytometry.
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