Antigen‐specific B cell detection reagents: Use and quality control

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Abstract
Tests for immunoglobulin reactivity with specific antigens are some of the oldest and most used assays in immunology. With efforts to understand B cell development, B cell dysregulation in autoimmunity, and to generate B cell vaccines for infectious agents, investigators have found the need to understand the ontogeny and regulation of epitope‐specific B cell responses. The synchrony between surface and secreted antibodies for individual B cells has led to the development of reagents and techniques to identify antigen‐specific B cells via reagent interactions with the B cell receptor complex. B cell antigen‐specific reagents have been reported for model systems of haptens, for whole proteins, and for identification of double stranded (ds) DNA antibody‐producing B cells using peptide mimics. Here we provide an overview of reported techniques for the detection of antigen‐specific B cell responses via secreted antibody or by the surface B cell receptor and briefly discuss our recent work developing a panel of reagents to probe the B cell response to HIV‐1 envelope. We also present an analysis of strengths and weaknesses of various methods for flow cytometric analysis of antigen‐specific B cells. © 2008 International Society for Advancement of Cytometry