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

J Immunol Methods. 2004 Mar;286(1-2):111-22. doi: 10.1016/j.jim.2003.12.015.


In the interest of better understanding the role of human memory B cells in protection against disease, we developed an assay to quantitate antigen-specific memory B cells in human blood. This assay utilizes a 6-day polyclonal stimulation of PBMC followed by an antigen-specific ELISPOT for the detection of memory B cells that have differentiated into antibody secreting cells (ASC) in vitro. We have used this assay to demonstrate that the anthrax vaccine (AVA; BioThrax) elicits a substantial population of protective-antigen (PA) specific memory B cells, and these B cells satisfy the canonical surface phenotype of human memory B cells: CD19(+)CD20(+)Ig(+)CD27(+). These anti-PA antigen-specific memory B cells are IgG(+) and represent up to 2% of circulating IgG(+) B cells. Furthermore, these results confirm that vaccine-elicited memory B cells reside in the CD27(+) B cell population. This ELISPOT-based system has been designed in a generalized manner, such that the assay can be rapidly adapted to detect human antigen-specific memory B cells of any given specificity. This method should be useful for quantitatively assessing the potency of vaccines and the longevity of B cell immunological memory to various vaccines or infectious diseases.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Anthrax Vaccines / immunology
  • Antibodies, Bacterial / blood
  • Antigens, Bacterial*
  • B-Lymphocytes / cytology
  • B-Lymphocytes / immunology*
  • Bacterial Toxins / immunology
  • Epitopes, B-Lymphocyte / immunology*
  • Flow Cytometry
  • Humans
  • Immunoenzyme Techniques / methods*
  • Immunologic Memory / immunology*
  • Immunomagnetic Separation
  • Immunophenotyping
  • Pokeweed Mitogens / immunology
  • Sensitivity and Specificity
  • Staphylococcus aureus / immunology


  • Anthrax Vaccines
  • Antibodies, Bacterial
  • Antigens, Bacterial
  • Bacterial Toxins
  • Epitopes, B-Lymphocyte
  • Pokeweed Mitogens
  • anthrax toxin