Blood from 3 healthy donors was used to separate peripheral blood mononuclear cells, using Ficoll-Paque separation media (GE Healthcare). MLR was essentially performed as previously described.38 (link) Further details are in the supplemental Material and methods. After 48 hours, all samples were treated for 5 hours with Golgi plug protein transport inhibitor, brefeldin (BD Biosciences) to inhibit cytokine accumulation in the Golgi complex and processed to measure interferon-γ (IFN-γ) production by CD4+ and CD8+ T cells by flow cytometry.
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Measuring IFN-γ production in T cells
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Corresponding Organization :
Other organizations : University of Siena, Sapienza University of Rome, Beth Israel Deaconess Medical Center, Harvard University
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Variable analysis
independent variables
- Separation of peripheral blood mononuclear cells using Ficoll-Paque separation media
- Interferon-γ (IFN-γ) production by CD4+ and CD8+ T cells
- Blood samples from 3 healthy donors
- Positive control: None mentioned
- Negative control: None mentioned
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