Vagaries of the ELISpot assay: Specific detection of antigen responsive cells requires purified CD8+ T cells and MHC class I expressing antigen presenting cell lines
YF Fuchs, GW Jainta, D Kühn, C Wilhelm, M Weigelt… - Clinical …, 2015 - Elsevier
Quantification of antigen-specific CD8+ T cells is important for monitoring infection,
vaccination, and response to therapy in cancer and immune-mediated diseases. Cytokine
enzyme-linked-immunospot (ELISpot) assays are often used for this purpose. We found that
substantial spot formation in IFNγ ELISpot assays occurred independently of CD8+ T cells
even when classical MHC class I restricted peptides are used for stimulation. Using
fractionated cells and intracellular cytokine staining, the non-CD8+ T cell IFNγ production …
vaccination, and response to therapy in cancer and immune-mediated diseases. Cytokine
enzyme-linked-immunospot (ELISpot) assays are often used for this purpose. We found that
substantial spot formation in IFNγ ELISpot assays occurred independently of CD8+ T cells
even when classical MHC class I restricted peptides are used for stimulation. Using
fractionated cells and intracellular cytokine staining, the non-CD8+ T cell IFNγ production …