title: Enzyme-linked immunospot assays for direct ex vivo measurement of vaccine-induced human humoral immune responses in blood.
abstract: The enzyme-linked immunospot (ELISPOT) assay was originally developed to enumerate antigen-specific antibody-secreting cells (ASCs), and has subsequently been adapted for various applications, including the detection cytokine-secreting cells. Owing to its exceptionally high sensitivity, the ELISPOT has proven to be especially useful for detecting discrete populations of active cells (e.g., antigen-specific cells). Because of its versatility, the ELISPOT assay is used for a wide range of applications, including clonal analyses of immune responses after vaccination or after immunotherapy. Here we describe standard protocols for the detection of human ASCs specific to virtually any vaccine antigen after enrichment of circulating plasmablasts. In addition, a protocol is described for the measurement of mucosal ASC responses after prior immunomagnetic enrichment of mucosally derived blood lymphocytes. The protocols described allow rapid (~6-8 h) detection of specific ASCs in small (1-2 ml) samples of blood and can be performed in resource-poor settings.
authors: Laboratory Sciences Division, International Vaccine Institute, Seoul, South Korea.
authors: Giulietta
authors: G
authors: Saletti
authors: Nicolas
authors: N
authors: Çuburu
authors: Jae Seung
authors: JS
authors: Yang
authors: Ayan
authors: A
authors: Dey
authors: Cecil
authors: C
authors: Czerkinsky
mesh_terms: D000921:Antibody-Producing Cells / Q000276:immunology*; D058501:Enzyme-Linked Immunospot Assay / Q000379:methods*; D006801:Humans; D056724:Immunity, Humoral / Q000276:immunology*; D009580:Nitroblue Tetrazolium; D014612:Vaccines / Q000097:blood / Q000276:immunology*
chemical_list: D014612:Vaccines; D009580:Nitroblue Tetrazolium
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