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. 2012 Dec;81(12):1019-21.
doi: 10.1002/cyto.a.22218. Epub 2012 Oct 18.

OMIP-014: validated multifunctional characterization of antigen-specific human T cells by intracellular cytokine staining

Stephen C De Rosa  1 , Donald K Carter, M Juliana McElrath
Affiliations

OMIP-014: validated multifunctional characterization of antigen-specific human T cells by intracellular cytokine staining

Stephen C De Rosa et al. Cytometry A. 2012 Dec.
No abstract available

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Figures

Figure 1
Figure 1
Example staining profile for PBMC stimulated with Staphylococcal enterotoxin B (SEB). The upper two rows show the gating hierarchy to identify CD4+ and CD8+ T cells. Due to pressure fluctuations during the first 10 seconds of collection of the sample using the high-throughput sampler (HTS), events collected during this time are excluded. The lower two rows show expression of the functional markers for CD4+ and CD8+ T cells. A gate is applied for each functional marker, not accounting for co-expression of other markers. Boolean gates are later created based on the gates shown to identify cells expressing various combinations of markers. Note that the functional gates for many markers are placed relatively high in reference to the negative cells to lower the number of cells falling into these gates for the unstimulated samples (not shown).
See this image and copyright information in PMC

References

    1. Horton H, Thomas EP, Stucky JA, Frank I, Moodie Z, Huang Y, Chiu YL, McElrath MJ, De Rosa SC. Optimization and validation of an 8-color intracellular cytokine staining (ICS) assay to quantify antigen-specific T cells induced by vaccination. J Immunol Methods. 2007;323:39–54. - PMC - PubMed
    1. Betts MR, Nason MC, West SM, De Rosa SC, Migueles SA, Abraham J, Lederman MM, Benito JM, Goepfert PA, Connors M, et al. HIV nonprogressors preferentially maintain highly functional HIV-specific CD8+ T cells. Blood. 2006;107:4781–9. - PMC - PubMed
    1. Betts MR, Brenchley JM, Price DA, De Rosa SC, Douek DC, Roederer M, Koup RA. Sensitive and viable identification of antigen-specific CD8+ T cells by a flow cytometric assay for degranulation. J Immunol Methods. 2003;281:65–78. - PubMed
    1. Chattopadhyay PK, Yu J, Roederer M. A live-cell assay to detect antigen-specific CD4+ T cells with diverse cytokine profiles. Nat Med. 2005;11:1113–7. - PubMed
    1. Mahnke YD, Roederer M. OMIP-001: Quality and phenotype of Ag-responsive human T-cells. Cytometry A. 2010;77A:819–20. - PubMed

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