Background information
IFN-γ (interferon-gamma) 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 TH 1 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 virus, CMV, EBV, HIV, HBV, and ADV.
Virus-specific T cells were expanded in vitro showing highly specific and very efficient killing of target cells and have been analyzed for TCR clonotypes.
The IFN-γ Secretion Assay was used for the isolation and analysis of antigen-specific T cells from PBMCs after stimulation with Tetanus Toxoid, minor histocompatibility antigens, and tumor antigens. The assay was also used to purify and analyze tumor-specific T cells from T cell lines, for the isolation of functional antigen-specific, IFN-γ-secreting T cells reacting to other tumor antigens, e.g. SSX, CEA, or HER2 from PBMCs or TILs (tumor infiltrating lymphocytes).
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.
The IFN-γ Secretion Assay was also used for isolation and functional characterization of allergen-specific T cells. Furthermore, the IFN-γ Secretion Assay was used for epitopemapping of MHC class II peptides.
IFN-γ-secreting human NK cells were isolated using the IFN-γ Secretion Assay. IFN-γ Secretion Assay reagents were reported to cross-react with chimpanzee cells but not with rhesus macaque cells.
The IFN-γ Secretion Assay can also be used for two-color cytokine analysis and allows counterstaining of peptide- MHC-tetramer-labeled cells. It can also be combined with flow cytometric proliferation assays.
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