Antigen-Reactive T Cell Enrichment for Direct, High-Resolution Analysis of the Human Naive and Memory Th Cell Repertoire
Abstract
Ag-specific CD4+ T cells orchestrating adaptive immune responses are crucial for the development of protective immunity, but also mediate immunopathologies. To date, technical limitations often prevented their direct analysis. In this study, we report a sensitive flow cytometric assay based on magnetic pre-enrichment of CD154+ T cells to visualize rare Ag-reactive naive and memory Th cells directly from human peripheral blood. The detection limit of ∼1 cell within 105–106 permitted the direct enumeration and characterization of auto-, tumor-, or neo-Ag–reactive T cells within the naive and even memory CD4+ T cell repertoire of healthy donors. Furthermore, the analysis of high target cell numbers after pre-enrichment of rare Ag-specific T cells from large blood samples dramatically improved the identification of small subpopulations. As exemplified in this work, the dissection of the Ag-specific memory responses into small cytokine-producing subsets revealed great heterogeneity between pathogens, but also pathogen-related microsignatures refining Th cell subset classification. The possibility to directly analyze CD4+ T cells reactive against basically any Ag of interest at high resolution within the naive and memory repertoire will open up new avenues to investigate CD4+ T cell–mediated immune reactions and their use for clinical diagnostics.
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CD154+ cells were magnetically enriched from 25 to 50 million PBMCs to detect T cells at low frequencies with a detection limit of 1 in 10^6.
“CD154 + cells were magnetically enriched from 25 to 50 million PBMCs to detect T cells at low frequencies (detection limit = 1 in 10 6 ). 4,42”
From Autoantigen-specific CD4+ T cells acquire an exhausted phenotype and persist in human antigen-specific autoimmune diseases (2024) · cites this paper as [42]
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Self-citation
- Authors
- Bacher, P., Schink, C., Teutschbein, J., Kniemeyer, O., Assenmacher, M., Brakhage, A. A., & Scheffold, A.
- Affiliations
- Miltenyi Biotec , 51429 Bergisch Gladbach,
- Miltenyi Biotec (Germany) (DE)
- Leibniz-Institut für Naturstoff-Forschung und Infektionsbiologie e. V. - Hans-Knöll-Institut (HKI) (DE)
- Jena University Hospital (DE)
- Charité – Universitätsmedizin Berlin (DE)
- Corresponding author
- Schink, Christian
- Assenmacher, Mario
- Venue
- The Journal of Immunology
- Publisher
- Oxford University Press (OUP)
- Published
- 2013-04
- Volume
- 190
- Issue
- 8
- Pages
- 3967-3976
- ISSN
- 0022-1767 (ISSN-L)
- 1550-6606
- OpenAlex
- W2100544695
- Cited by
- 173
- References
- 38
- Citations by year
- 2026: 7 · 2025: 11 · 2024: 14 · 2023: 11 · 2022: 19 · 2021: 20 · 2020: 23 · 2019: 14 · 2018: 16 · 2017: 11 · 2016: 5 · 2015: 12 · 2014: 7 · 2013: 3
- FWCI
- 4.97
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- The Journal of Immunology (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
- Life Sciences › Immunology and Microbiology › Immunology › Immunotherapy and Immune Responses
- Life Sciences › Immunology and Microbiology › Immunology › Psoriasis: Treatment and Pathogenesis
- Keywords
- antigen-specific T cells, CD4+ T cells, naive T cells, memory T cells, CD154, flow cytometry assay, T cell receptor repertoire, neoantigens, peripheral blood
- MeSH terms
- Antigen-Antibody Reactions, Antigen-Presenting Cells, Aspergillus fumigatus, Cell Differentiation, Cell Line, Clone Cells, T-Lymphocytes, Helper-Inducer, Humans, Immunologic Memory, T-Lymphocyte Subsets, Antigen Presentation, CD4 Lymphocyte Count, Epitopes, T-Lymphocyte
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