PBMCs were isolated from whole blood by Ficoll-Paque density gradient centrifugation (GE Healthcare Bio-sciences), as described previously (55 (link)). CD8+ T cells were subsequently isolated from PBMCs by negative selection using the EasySep Human CD8+ T Cell Enrichment Kit (STEMCELL Technologies Inc.) according to the manufacturer’s instructions. The CD8+ T cells were maintained in RPMI-1640 medium supplemented with 10% human serum, 200 u/ml penicillin, 200 µg/ml streptomycin, 1 mM L-glutamine, and 10 mM HEPES (55 (link)). Cells were activated for 72–96 h in a cell culture dish coated with 10 µg/ml mouse anti-human CD3 antibody (Biolegend) and 10 µg/ml mouse anti-human CD28 antibody (Biolegend).
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CD8+ T Cell Isolation and Activation
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Corresponding Organization : University of Cincinnati
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Variable analysis
independent variables
- Activating CD8+ T cells with 10 µg/ml mouse anti-human CD3 antibody and 10 µg/ml mouse anti-human CD28 antibody
- Not explicitly mentioned
- Maintaining CD8+ T cells in RPMI-1640 medium supplemented with 10% human serum, 200 u/ml penicillin, 200 µg/ml streptomycin, 1 mM L-glutamine, and 10 mM HEPES
- Isolating CD8+ T cells from PBMCs by negative selection using the EasySep Human CD8+ T Cell Enrichment Kit
- Positive control: Activating CD8+ T cells with anti-CD3 and anti-CD28 antibodies
- Negative control: Not explicitly mentioned
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