Preparation and analysis of antigen-specific memory B cells

E. Yefenof, V. M. Sanders, E. C. Snow, R. J. Noelle, K. G. Oliver, J. W. Uhr, E. S. Vitetta

Research output: Contribution to journalArticlepeer-review

32 Scopus citations

Abstract

A procedure has been developed for the enrichment of TNP-binding memory B cells (TNP-MABC) from spleens of immunized mice. More than 75% of the cells expressed surface IgM (sIgM) and IgD (sIgD) and about 9% expressed surface IgG (sIgG). The TNP-MABC consisted of small resting lymphocytes with high affinity antigen-binding receptors. These cells expressed increased densities of Ia antigens and decreased densities of sIgD. Adoptive transfer of the cells into irradiated, carrier-primed syngeneic recipients resulted in their differentiation into IgG anti-TNP antibody-secreting cells. TNP-MABC secreted high affinity IgG anti-TNP antibodies when cultured in vitro with carrier-primed T cells and antigen. Limiting dilution analysis revealed that TNP-MABC contained a relatively low frequency of precursors for IgG-secreting cells that had an exceptionally large clone size. These results show that a highly enriched population of antigen-specific memory B cells can now be prepared and used to analyze their activation requirements.

Original languageEnglish (US)
Pages (from-to)3777-3784
Number of pages8
JournalJournal of Immunology
Volume135
Issue number6
StatePublished - 1985

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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