Peripheral blood was obtained from healthy donors through an IRB-approved program at the University of Maryland, Baltimore. Peripheral blood mononuclear cells (PBMCs) were harvested from peripheral blood by Ficoll–Paque (GE Healthcare) gradient centrifugation.
T cells from PBMCs were isolated using human Pan T Cell Isolation Kits (Miltenyi Biotec) according to the manufacturer’s protocol. T cells were further cultured with T-cell medium, which was composed of RPMI 1640 (Life Technologies), 2 mMl -glutamine (Life Technologies), 10% FBS (Benchmark), and 100 U/ml pen/strep (Life Technologies)] supplemented with recombinant IL-2 (rIL-2) (30 U/ml) (Millipore Sigma). Human T-Activator CD3/CD28 Dynabeads™ (Thermo Fisher Scientific) were used for T cell expansion and activation according to the manufacturer’s protocol. Activated T cells were incubated at 37 °C and 5% CO2 for 7 days before injection into mice.
Before injection into mice, T cells were labeled by incubation with 320 μg/mL Xenolight Dir (PerkinElmer) for 30 min and subsequently washed twice with chilled PBS.
CD14+ cells and CD16+ cells were isolated by using human CD14 microbeads (Miltenyi Biotec) and CD16 microbeads (Miltenyi Biotec), respectively, according to the manufacturer’s protocol.
T cells from PBMCs were isolated using human Pan T Cell Isolation Kits (Miltenyi Biotec) according to the manufacturer’s protocol. T cells were further cultured with T-cell medium, which was composed of RPMI 1640 (Life Technologies), 2 mM
Before injection into mice, T cells were labeled by incubation with 320 μg/mL Xenolight Dir (PerkinElmer) for 30 min and subsequently washed twice with chilled PBS.
CD14+ cells and CD16+ cells were isolated by using human CD14 microbeads (Miltenyi Biotec) and CD16 microbeads (Miltenyi Biotec), respectively, according to the manufacturer’s protocol.