Anti human cd3 cd28 antibodies

Whole blood from HNSCC patients and HDs was obtained and peripheral blood mononuclear cells were isolated using Ficoll-paque PLUS density gradient (GE Healthcare Biosciences) as described previously.⁶ (link) CD3⁺, CD8⁺, and CD8⁺ memory T cells were each isolated by negative selection using commercially available kits (EasySep Enrichment kits, StemCell Technologies). The purities of these enriched T cell populations were determined using flow cytometry as indicated in Figure S3. T cells were maintained in T cell medium (RPMI-1640 medium containing L-glutamine supplemented with 200 U/mL penicillin, 200 mg/mL streptomycin, 1 mM L-glutamine, 10 mM HEPES, and 10% human serum) and then either remained unstimulated for 12–24 h (±NPs) or were activated in cell culture plates coated with 10 μg/mL anti-human CD3/CD28 antibodies (BioLegend) or 25 μL/mL ImmunoCult Human CD3/CD28 T Cell Activator (StemCell Technologies) for 72–96 h (±NPs) in the 37°C/5% CO₂ incubator as indicated in the following protocols.