Journal
NATURE PROTOCOLS
Volume 8, Issue 6, Pages 1073-1087Publisher
NATURE PORTFOLIO
DOI: 10.1038/nprot.2013.058
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Funding
- Bill & Melinda Gates Foundation [OPP1017157, PROVIDE]
- European Union Seventh Framework Program [280873 ADITEC]
- World Health Organization (WHO
- Global Polio Eradication Program)
- PATH Vaccine Solutions (EVI Program)
- Institut National de la Sante et de la Recherche Medicale (INSERM, France)
- Swedish Agency for International Cooperation (SIDA)
- Swedish National Research Council
- government of the Republic of Korea
- government of the Republic of Sweden
- government of the Republic of the Netherlands
- Bill and Melinda Gates Foundation [OPP1017157] Funding Source: Bill and Melinda Gates Foundation
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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 (similar to 6-8 h) detection of specific ASCs in small (1-2 ml) samples of blood and can be performed in resource-poor settings.
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