| Description |
| The IFN-γ Secretion Assay - Cell Enrichment and Detection Kit was developed for the sensitive detection as well as the enrichment of human IFN-γ-secreting cells. |
| Details |
Background information
Interferon gamma (IFN-γ) is predominantly secreted by activated CD8+ and CD4+ memory and effector T cells and by NK cells. It is mainly involved in the regulation of inflammatory immune responses. These T1 types of immune mechanisms are effective against intracellular pathogens and tumors. IFN-γ-secreting T cells can also be involved in immunological disorders such as autoimmune reactions.
Downstream applications
Virus-specific T cells were investigated after stimulation with peptides or proteins derived from influenza virus1, CMV2,3, EBV4,5,6, HIV5,7,8,9,10, HBV11, and ADV24.
Virus-specific T cells were expanded in vitro1,2,3,5,6,9 showing highly specific and very efficient killing of target cells and have been analyzed for TCR clonotypes8,10.
IFN-γ Secretion Assays were used for the isolation and analysis of antigen-specific T cells from PBMCs after stimulation with Tetanus Toxoid1, minor histocompatibility antigens, and tumor antigens12,13,14,15. IFN-γ Secretion Assays were also used to purify and analyze tumor-specific T cells from T cell lines13,14 and for isolation of functional antigen-specific, IFN-γ-secreting T cells reacting to other tumor antigens, e.g., SSX14, CEA16, or HER217 from PBMCs or TILs (tumor infiltrating lymphocytes)14,18.
The IFN-γ Secretion Assay was used to counterstain peptide-MHC-tetramer-labeled Melan A-specific CD8+ T cells to analyze functionality of the tetramer-positive cells.12,15
The IFN-γ Secretion Assay was also used for isolation and functional characterization of allergen-specific T cells.21 Furthermore, the IFN-γ Secretion Assay was used for epitope mapping of MHC class II peptides.19
IFN-γ-secreting human NK cells were isolated using the IFN-γ Secretion Assay.20 IFN-γ Secretion Assay reagents were reported to cross-react with chimpanzee cells22, but not with rhesus macaque cells.
The IFN-γ Secretion Assays can also be used for two-color cytokine analysis9 and allows counterstaining of peptide-MHC-tetramer-labeled cells 12,15. It can also be combined with flow cytometric proliferation assays 23. |
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| Figure 1 |
| PBMCs from a CMV+ donor were stimulated with PepTivator™ CMV pp65 or left untreated. Cells were stained and isolated according to secretion of IFN-γ using the IFN-γ Secretion Assay - Cell Enrichment and Detection Kit. The cells were counterstained with CD8-FITC. Cell debris and dead cells were excluded from the analysis based on scatter signals and PI fluorescence. |
| Stimulated sample |
| A: Before enrichment |
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| B: After enrichment |
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| Unstimulated control |
| C: Before enrichment |
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| D: After enrichment |
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