Profiling antibody responses by multiparametric analysis of primary B cells

Craig M. Storya, Eliseo Papa, Chih Chi Andrew Hu, Jehnna L. Ronan, Kara Herlihy, Hidde L. Ploegh, J. Christopher Love

Research output: Contribution to journalArticle

53 Scopus citations

Abstract

Determining the efficacy of a vaccine generally relies on measuring neutralizing antibodies in sera. This measure cannot elucidate the mechanisms responsible for the development of immunological memory at the cellular level, however. Quantitative profiles that detail the cellular origin, extent, and diversity of the humoral (antibody-based) immune response would improve both the assessment and development of vaccines. Here, we describe a novel approach to collect multiparametric datasets that describe the specificity, isotype, and apparent affinity of the antibodies secreted from large numbers of individual primary B cells (≈103-104). The antibody/antigen binding curves obtained by this approach can be used to classify closely related populations of cells using algorithms for data clustering, and the relationships among populations can be visualized graphically using affinity heatmaps. The technique described was used to evaluate the diversity of antigen-specific antibody-secreting cells generated during an in vivo humoral response to a series of immunizations designed to mimic a multipart vaccination. Profiles correlating primary antibody-producing cells with the molecular characteristics of their secreted antibodies should facilitate both the evaluation of candidate vaccines and, broadly, studies on the repertoires of antibodies generated in response to infectious or autoimmune diseases.

Original languageEnglish (US)
Pages (from-to)17902-17907
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume105
Issue number46
DOIs
StatePublished - Nov 18 2008

Keywords

  • Affinity
  • Immunomics
  • Microengraving
  • Soft lithography
  • Vaccines

ASJC Scopus subject areas

  • General

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