Clonal dissection of the human memory B-cell repertoire following infection and vaccination

Eur J Immunol. 2009 May;39(5):1260-70. doi: 10.1002/eji.200839129.


The analysis of the human memory B-cell repertoire is of both fundamental and practical significance. We developed a simple method for the selective activation of memory B cells in total fresh or frozen PBMC using a combination of R848 and IL-2. In these conditions, 30-40% of memory B cells generated clones producing on average 200 ng IgG in 10 days. This method was used to measure the frequency of antigen-specific memory B cells as well as the fine specificity, cross-reactivity and neutralizing activity of the secreted antibodies. Following influenza vaccination, specific B cells expanded dramatically, reaching up to 50% of total clonable memory B cells on day 14. Specific B-cell expansions were detected also in individuals that did not show a significant serological response. Dynamic changes and persistence of B cells specific for a variety of pathogens were documented in serial PBMC samples collected over almost two decades. These results reveal novel aspects of memory B-cell kinetics and provide a powerful tool to monitor immune responses following infection and vaccination.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • B-Lymphocytes / immunology*
  • Cell Differentiation / drug effects
  • Cell Differentiation / immunology
  • Clone Cells / immunology
  • Cytokines / pharmacology
  • Enzyme-Linked Immunosorbent Assay
  • Epitopes, B-Lymphocyte / immunology
  • Flow Cytometry
  • Humans
  • Immunoglobulin G / immunology
  • Immunologic Memory / immunology*
  • Influenza A Virus, H1N1 Subtype / immunology*
  • Influenza Vaccines / immunology*
  • Influenza, Human / blood
  • Influenza, Human / immunology*
  • Influenza, Human / virology
  • Kinetics
  • Leukocytes, Mononuclear / immunology
  • Toll-Like Receptors / agonists
  • Toll-Like Receptors / immunology


  • Cytokines
  • Epitopes, B-Lymphocyte
  • Immunoglobulin G
  • Influenza Vaccines
  • Toll-Like Receptors