Inguinal, axillary, brachial and mesenteric lymph nodes (LN) or spleens were isolated from CD45.1 OT-I or P14 LCMV mice. LN and spleens were meshed through 70μm filters and treated with ACK red blood cell lysis buffer. CD8+ T cells were purified using EasySep CD8 negative selection kits (Stemcell Technologies). 1×105 (for >14 day read-out) – 2×106 T cells (for day 4 read-out) were adoptively transferred through retro-orbital injection in 100μl PBS.
For the comparison of OT-I T cells and p14 LCMV T cells, mice received a 1:1 mix of both T cells in 100μl of PBS through retro-orbital injection. The following day mice were inoculated with a bolus of CFA containing gp33-peptide (50μg/mouse; Anaspec) and SL8/SIINFEKL peptide (50μg/mouse; Anaspec) subcutaneously, to sustain both T cell populations.
For the comparison of OT-I T cells and p14 LCMV T cells, mice received a 1:1 mix of both T cells in 100μl of PBS through retro-orbital injection. The following day mice were inoculated with a bolus of CFA containing gp33-peptide (50μg/mouse; Anaspec) and SL8/SIINFEKL peptide (50μg/mouse; Anaspec) subcutaneously, to sustain both T cell populations.