Reagent Tracker Platform Verifies and Provides Audit Trails for the Error-Free Implementation of T-Cell ImmunoSpot® Assays

Methods Mol Biol. 2024:2768:105-115. doi: 10.1007/978-1-0716-3690-9_7.

Abstract

ELISPOT and FluoroSpot assays, collectively called ImmunoSpot assays, permit to reliable detection of rare antigen-specific T cells in freshly isolated cell material, such as peripheral blood mononuclear cells (PBMC). Establishing their frequency within all PBMC permits to assess the magnitude of antigen-specific T-cell immunity; the simultaneous measurement of their cytokine signatures reveals these T-cells' lineage and effector functions, that is, the quality of T-cell-mediated immunity. Because of their unparalleled sensitivity, ease of implementation, robustness, and frugality in PBMC utilization, T-cell ImmunoSpot assays are increasingly becoming part of the standard immune monitoring repertoire. For regulated workflows, stringent audit trails of the data generated are a requirement. While this has been fully accomplished for the analysis of T-cell ImmunoSpot assay results, such are missing for the wet laboratory implementation of the actual test performed. Here we introduce a solution for enhancing and verifying the error-free implementation of T-cell ImmunoSpot assays.

Keywords: ELISPOT; FluoroSpot; High throughput; Immune memory; Immune monitoring; ImmunoSpot®; T cells.

MeSH terms

  • Cytokines
  • Enzyme-Linked Immunospot Assay / methods
  • Immunity, Cellular
  • Leukocytes, Mononuclear*
  • T-Lymphocytes*

Substances

  • Cytokines