Naïve T cells from TcR Tg mice were obtained from pooled spleen and lymph nodes as previously described (25 (link)). Resulting TcR Tg cells were routinely >97% TcR+ and expressed a characteristic naive phenotype (small size, CD62Lhi, CD44lo and CD25lo). TH1 or Tc1 effectors were generated from naïve TcR Tg T cells cultured as previously described (25 (link)–27 (link)). In vitro-generated memory cells were obtained from effector cultures that were washed several times and rested for at least 3 days in media free of antigen (Ag) or exogenous cytokine. In some experiments, cells were labeled with carboxyfluorescein succinimidyl ester (CFSE) (Molecular Probes) prior to adoptive transfer. All cell populations were adoptively transferred to unprimed mice in 200 μl PBS by i.v. injection.
Mice were injected i.p. with IL-6-neutalizing Ab (MP5-20F3), or isotype control Ab (HRPN) (BioXcell) as indicated. In some experiments, mice were treated i.p. with 0.5 mg of anti Ly-6G Ab to deplete neutrophils (1A8) or isotype control (2A3) (BioXcell) on 0, 2, 4, and 6 dpi.
Mice were injected i.p. with IL-6-neutalizing Ab (MP5-20F3), or isotype control Ab (HRPN) (BioXcell) as indicated. In some experiments, mice were treated i.p. with 0.5 mg of anti Ly-6G Ab to deplete neutrophils (1A8) or isotype control (2A3) (BioXcell) on 0, 2, 4, and 6 dpi.
