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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.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
|August 3, 2020
PubMed

Insights

This study presents a 24-color flow cytometry panel for detailed B-cell analysis in chronic infections. The panel aids in characterizing antigen-specific B cells and B-cell subsets, applicable to autoimmunity research.

Area of Science:

  • Immunology
  • Cell Biology
  • Medical Research

Background:

  • Chronic infections and autoimmune diseases involve complex B-cell dynamics.
  • Characterizing B-cell subsets and antigen-specific responses is crucial for understanding disease pathogenesis.
  • Existing flow cytometry panels may lack the comprehensive markers needed for in-depth B-cell analysis.

Purpose of the Study:

  • To develop and optimize a 24-color flow cytometry panel for detailed characterization of B-cell subsets.
  • To enable precise delineation of antigen-specific B cells in the context of chronic infections, such as hepatitis B virus (HBV).
  • To provide a versatile platform adaptable for studying other chronic diseases and research questions.

Main Methods:

  • Optimization of a 24-color flow cytometry panel using cryopreserved human peripheral blood mononuclear cells (PBMCs).
  • Selection of markers to distinguish B-cell lineages (e.g., CD19, CD20, CD10, CD38, CD24, IgM, IgD, CD27, CD21, CD43, CD5).
  • Inclusion of antigen-specific probes (initially for HBV) and a comprehensive set of functional, regulatory, and inhibitory markers (e.g., CXCR3, CXCR5, CD86, CD32, BTLA, CD39, PD-1, FcRL5, CD11c, CD22).

Main Results:

  • A validated 24-color flow cytometry panel capable of extensive B-cell subset characterization.
  • Demonstrated ability to identify and analyze antigen-specific B cells, exemplified by HBV-specific B cells.
  • The panel incorporates functional markers for in-depth analysis of B-cell states during chronic infection.

Conclusions:

  • The developed flow cytometry panel provides a robust tool for dissecting B-cell heterogeneity in chronic infections and autoimmunity.
  • This versatile panel can be customized with different antigen-specific probes or markers for diverse research applications.
  • Enables advanced investigation into the role of B cells in disease pathogenesis and potential therapeutic targets.

Keywords:
B cellsHBVantigen-specific B cellschronic Infectionsmemory B cellsphenotyping

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