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.