Adoptive transfer of OT-1 T cells

Spleen and MLNs were collected from OT-1 TCR-Tg Rag1 knockout CD45.1 mice. OT-1 T cells were isolated using a CD8⁺ T cell Isolation kit (Miltenyi Biotec) according to the manufacturer’s protocol. The purity of OT-1 T cells was confirmed to be >90%, which was evaluated by flow cytometry using anti-Vα2 TCR (BD Biosciences) and anti-CD45.1 (BD Biosciences). 50,000 OT-1 T cells in 200 μL PBS were injected into the tail vein of each recipient mouse 1 day before immunization. In some experiments, OT-1 T cells were labeled with CFSE (Invitrogen) prior to the adoptive transfer. Mice were sacrificed at indicated time points, single-cell suspensions were obtained from lung and MLNs for flow cytometry.