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@ARTICLE{Schllhorn:265700,
      author       = {A. Schöllhorn and A. Maia and F. Kimmerle and J. Born and
                      H.-G. Rammensee$^*$ and S. Dimitrov and C.
                      Gouttefangeas$^*$},
      title        = {{S}taining of activated ß2-integrins in combination with
                      {CD}137 and {CD}154 for sensitive identification of
                      functional antigen-specific {CD}4+ and {CD}8+ {T} cells.},
      journal      = {Frontiers in immunology},
      volume       = {13},
      issn         = {1664-3224},
      address      = {Lausanne},
      publisher    = {Frontiers Media},
      reportid     = {DKFZ-2023-00295},
      pages        = {1107366},
      year         = {2023},
      abstract     = {Common flow cytometry-based methods used for functional
                      assessment of antigen-specific T cells rely on de novo
                      expression of intracellular cytokines or cell surface
                      activation induced markers. They come with some limitations
                      such as complex experimental setting, loss of cell viability
                      and often high unspecific background which impairs assay
                      sensitivity. We have previously shown that staining of
                      activated ß2-integrins either with multimers of their
                      ligand ICAM-1 or with a monoclonal antibody can serve as a
                      functional marker detectable on T cells after minutes (CD8+)
                      or few hours (CD4+) of activation. Here, we present a simple
                      method for detection of activated ß2-integrins in
                      combination with established cell surface activation induced
                      markers. We observed that activated ß2-integrins were still
                      detectable after 14 hours of stimulation, allowing their
                      detection together with CD137 and CD154. Combinatorial
                      gating of cells expressing activated ß2-integrins and CD137
                      or CD154 reduced background in unstimulated samples,
                      increasing the signal-to-noise ratio and allowing improved
                      assessment of low-frequency T cell responses. Extracellular
                      staining of these markers highly correlated with production
                      of intracellular cytokines IL-2, TNF or IFNγ in CD4+ and
                      CD8+ T cells. As an exemplary application, SARS-CoV-2
                      spike-specific T cell responses were assessed in individuals
                      after COVID-19 vaccination. This method should be useful for
                      epitope discovery projects and for the simultaneous
                      monitoring of low-frequency antigen-specific CD4+ and CD8+ T
                      cell responses in various physiological situations.},
      keywords     = {Humans / CD8-Positive T-Lymphocytes / CD4-Positive
                      T-Lymphocytes / Integrins: metabolism / COVID-19 Vaccines:
                      metabolism / COVID-19: metabolism / SARS-CoV-2 / Antigens:
                      metabolism / CD40 Ligand / Cytokines: metabolism / CD137
                      (Other) / CD154 (Other) / CD4+ T cells (Other) / CD8+ T
                      cells (Other) / activation induced marker (AIM) (Other) /
                      antigen-specificity (Other) / cell surface staining (Other)
                      / integrin activation (Other) / Integrins (NLM Chemicals) /
                      COVID-19 Vaccines (NLM Chemicals) / Antigens (NLM Chemicals)
                      / CD40 Ligand (NLM Chemicals) / Cytokines (NLM Chemicals)},
      cin          = {TU01},
      ddc          = {610},
      cid          = {I:(DE-He78)TU01-20160331},
      pnm          = {899 - ohne Topic (POF4-899)},
      pid          = {G:(DE-HGF)POF4-899},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:36741378},
      pmc          = {pmc:PMC9892897},
      doi          = {10.3389/fimmu.2022.1107366},
      url          = {https://inrepo02.dkfz.de/record/265700},
}