A sensitive and less cytotoxic assay for identification of proliferating T cells based on bioorthogonally-functionalized uridine analogue

Stempels, Femmy C.; Wit, A S de; Swierstra, M S; Maassen, Sjors; Bianchi, Frans; Bogaart, Geert van den; Baranov, Maksim V.

doi:10.1016/j.jim.2022.113228

Cited by 1 publication

(2 citation statements)

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“…Traditionally, antigen‐specific T‐cell responses have been quantified by measuring cell proliferation, cytokine production and/or peptide–major histocompatibility complex (MHC) multimer staining. Proliferation assays are typically performed by stimulating peripheral blood mononuclear cells (PBMCs) with an antigen of interest, and quantifying cell proliferation by H 3 ‐thymidine uptake or dilution of a cell‐proliferation dye, such as carboxyfluorescein succinimidyl ester (CFSE) after 4–7 days 1,2 . However, these assays require antigen stimulation that directly affects the phenotype of proliferating cells, meaning the initial frequency of antigen‐specific T cells in the original sample cannot be quantified.…”

Section: Introductionmentioning

confidence: 99%

“…Proliferation assays are typically performed by stimulating peripheral blood mononuclear cells (PBMCs) with an antigen of interest, and quantifying cell proliferation by H 3 -thymidine uptake or dilution of a cell-proliferation dye, such as carboxyfluorescein succinimidyl ester (CFSE) after 4-7 days. 1,2 However, these assays require antigen stimulation that directly affects the phenotype of proliferating cells, meaning the initial frequency of antigenspecific T cells in the original sample cannot be quantified. Proliferation assays also do not detect hypoproliferative antigen-specific cells, such as regulatory T cells (Tregs), and are influenced by bystander activation, whereby interleukin (IL)-2 secreted by antigen-specific T cells drives antigenindependent proliferation of nearby T cells.…”

Section: Introductionmentioning

confidence: 99%

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T‐cell activation–induced marker assays in health and disease

et al. 2023

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Activation‐induced marker (AIM) assays have proven to be an accessible and rapid means of antigen‐specific T‐cell detection. The method typically involves short‐term incubation of whole blood or peripheral blood mononuclear cells with antigens of interest, where autologous antigen‐presenting cells process and present peptides in complex with major histocompatibility complex (MHC) molecules. Recognition of peptide–MHC complexes by T‐cell receptors then induces upregulation of activation markers on the T cells that can be detected by flow cytometry. In this review, we highlight the most widely used activation markers for assays in the literature while identifying nuances and potential downfalls associated with the technique. We provide a summary of how AIM assays have been used in both discovery science and clinical studies, including studies of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) immunity. This review primarily focuses on AIM assays using human blood or peripheral blood mononuclear cell samples, with some considerations noted for tissue‐derived T cells and nonhuman samples. AIM assays are a powerful tool that enables detailed analysis of antigen‐specific T‐cell frequency, phenotype and function without needing to know the precise antigenic peptides and their MHC restriction elements, enabling a wider analysis of immunity generated following infection and/or vaccination.

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Section: Introductionmentioning

confidence: 99%