Cyto 2011—Overview of the XXVI ISAC Congress Proceedings Issue of Cytometry

Smith, Paul J.; Radbruch, Andreas

doi:10.1002/cyto.a.21069

Cited by 2 publications

(3 citation statements)

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“…Our improved method was connected to reduced cell loss, as well as efficient debarcoding and immunophenotyping of low-frequency antigen-specific T cells by means of mass cytometry. The latter technology is advantageous over spectral cytometry when it comes to assessment of multiple cytokines and/or transcription factors as part of the assay’s marker panel ( Smith et al., 2023 ).…”

Section: Discussionmentioning

confidence: 99%

“…Both CyTOF and spectral cytometry overcome technical limitations associated to autofluorescence related to specific cell types. However, for functional studies requiring detection of multiple cytokines or transcription factors, CyTOF appears superior ( Smith et al., 2023 ). During the last years, CyTOF has significantly advanced scientific fields related to cancer ( Levine et al., 2015 ; Spitzer et al., 2015 ; Krieg et al., 2018 ), allergy, autoimmunity ( Hartmann et al., 2016 ; Rao et al., 2017 ; Mrdjen et al., 2018 ), and infection and hematopoiesis ( Bendall et al., 2011 ; Bendall et al., 2014 ) by providing detailed insights into several research questions.…”

Section: Introductionmentioning

confidence: 99%

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A novel mass cytometry protocol optimized for immunophenotyping of low-frequency antigen-specific T cells

Balz,

Grange,

Pegel

et al. 2024

Front. Cell. Infect. Microbiol.

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Understanding antigen-specific T-cell responses, for example, following virus infections or allergen exposure, is of high relevance for the development of vaccines and therapeutics. We aimed on optimizing immunophenotyping of T cells after antigen stimulation by improving staining procedures for flow and mass cytometry. Our method can be used for primary cells of both mouse and human origin for the detection of low-frequency T-cell response using a dual-barcoding system for individual samples and conditions. First, live-cell barcoding was performed using anti-CD45 antibodies prior to an in vitro T-cell stimulation assay. Second, to discriminate between stimulation conditions and prevent cell loss, sample barcoding was combined with a commercial barcoding solution. This dual-barcoding approach is cell sparing and, therefore, particularly relevant for samples with low cell numbers. To further reduce cell loss and to increase debarcoding efficiency of multiplexed samples, we combined our dual-barcoding approach with a new centrifugation-free washing system by laminar flow (Curiox™). Finally, to demonstrate the benefits of our established protocol, we assayed virus-specific T-cell response in SARS-CoV-2–vaccinated and SARS-CoV-2–infected patients and compared with healthy non-exposed individuals by a high-parameter CyTOF analysis. We could reveal a heterogeneity of phenotypes among responding CD4, CD8, and gd-T cells following antigen-specific stimulations. Our protocol allows to assay antigen-specific responses of minute populations of T cells to virus-derived peptides, allergens, or other antigens from the same donor sample, in order to investigate qualitative and quantitative differences.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A novel mass cytometry protocol optimized for immunophenotyping of low-frequency antigen-specific T cells

Balz,

Grange,

Pegel

et al. 2024

Front. Cell. Infect. Microbiol.

View full text Add to dashboard Cite

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“…These unidirectional HAPS (uHAPs) are reduced in hypoxia to the metabolically stable reduction products (AQ4 and OCT1001, respectively). Studies with AQ4N and OCT1002 show that hypoxia‐mediated reduction results in a product with high affinity for DNA and targeting of topoisomerase II,which can effect a long‐term inhibition of both DNA replication and G2/M cell transition …”

Section: Introductionmentioning

confidence: 99%