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Related Experiment Videos

Tracking human antigen-specific memory B cells: a sensitive and generalized ELISPOT system.

Shane Crotty1, Rachael D Aubert, John Glidewell

  • 1Emory Vaccine Center and Department of Microbiology and Immunology, Emory University School of Medicine, 1510 Clifton Road, Rm G-211, Atlanta, GA 30322, USA. shane@liai.org

Journal of Immunological Methods
|April 17, 2004
PubMed
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We developed a new assay to quantify antigen-specific memory B cells in human blood. This method successfully detected protective-antigen specific memory B cells elicited by the anthrax vaccine.

Area of Science:

  • Immunology
  • Vaccinology
  • Cell Biology

Background:

  • Understanding the role of human memory B cells is crucial for disease protection.
  • Existing methods for quantifying antigen-specific memory B cells are limited.
  • Memory B cells are key for long-term immunity following vaccination or infection.

Purpose of the Study:

  • To develop and validate a novel assay for quantifying antigen-specific memory B cells in human blood.
  • To assess the immunogenicity of the anthrax vaccine (AVA; BioThrax) in terms of memory B cell response.
  • To establish a generalized method for detecting memory B cells specific to various antigens.

Main Methods:

  • A 6-day polyclonal stimulation of peripheral blood mononuclear cells (PBMC).
  • Antigen-specific ELISPOT assay to detect memory B cells differentiating into antibody-secreting cells (ASC) in vitro.

Related Experiment Videos

  • Characterization of memory B cell surface phenotype (CD19(+)CD20(+)Ig(+)CD27(+)).

    • Main Results:

    • The anthrax vaccine elicits a significant population of protective-antigen (PA) specific memory B cells.
    • These vaccine-induced memory B cells exhibit the canonical human memory B cell phenotype (CD19(+)CD20(+)Ig(+)CD27(+)) and are IgG+.
    • Anti-PA specific memory B cells constitute up to 2% of circulating IgG+ B cells, residing within the CD27+ population.

    Conclusions:

    • The developed ELISPOT-based assay is effective in quantifying antigen-specific memory B cells in human blood.
    • The anthrax vaccine induces a robust and phenotypically defined memory B cell response.
    • This adaptable assay can be used to assess vaccine potency and the longevity of B cell memory against various pathogens.