title: ELISPOT cell rescue.
abstract: The enzyme-linked immunospot (ELISPOT) assay is a highly sensitive and reproducible method for quantifying T cell-mediated immune responses, and has been used to measure antigen-specific responses post-vaccination. While there are several advantages of the ELISPOT assay for use in field settings for large-scale vaccination trials, blood draw volumes are often limited, and the number of antigen-specific responses that can be measured is constrained by the limited cell number. We reasoned that it should be possible to salvage and rescue viable cells from a completed ELISPOT assay post-incubation, to use for further experimentation. Here, we show that cells rescued from an ELISPOT plate after assay are viable, and may be used in a second cytokine-producing assay, in a proliferation assay, or to provide a source of DNA for genetic studies such as human leukocyte antigen (HLA) typing. Rescue of cells after an ELISPOT assay will be particularly useful for increasing sample utility and maximizing data collection from T cell assays in vaccine trials.
authors.affiliation: Gladstone Institute of Virology and Immunology, University of California, PO Box 419100, San Francisco, CA 94141-9100, USA.
authors.forename: Duncan A
authors.initials: DA
authors.lastname: Meiklejohn
authors.forename: R Karl
authors.initials: RK
authors.lastname: Karlsson
authors.forename: Annika C
authors.initials: AC
authors.lastname: Karlsson
authors.forename: Joan M
authors.initials: JM
authors.lastname: Chapman
authors.forename: Douglas F
authors.initials: DF
authors.lastname: Nixon
authors.forename: Becky
authors.initials: B
authors.lastname: Schweighardt
mesh_terms: D004247:DNA / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006680:HLA Antigens / Q000235:genetics* / Q000276:immunology; D006801:Humans; D007371:Interferon-gamma / Q000032:analysis / Q000276:immunology; D007962:Leukocytes / Q000276:immunology* / Q000378:metabolism
chemical_list: D006680:HLA Antigens; D007371:Interferon-gamma; D004247:DNA