An MCA205 fibrosarcoma cell line expressing OVA protein was previously established21 (link). RMA cells, a Rauscher MuLV-induced lymphoma21 (link) was kindly provided by Dr Akira Shibuya (University of Tsukuba, Japan). Mice were inoculated subcutaneously with 8 × 105 MCA-OVA or 1 × 105 RMA, or were intravenously injected with 1 × 105 of the OVA/firefly luciferase-expressing melanoma cell line, MO4-Luc26 (link)52 (link), in the pulmonary metastatic model. Tumour size is expressed as a tumour index, the square root of (length × width), as described previously53 (link). To measure MO4 metastasis in the lung, luminescence images were analysed using NightOWL II (Berthold Technologies)26 (link). Two hundred μg of anti-IL-6R Ab, 15A7; anti-IL-6 Ab, MP5–20F3 (BioXCell); or control rat IgG Ab (Millipore) was injected 1 day before and after immunization. Daily intraperitoneal injections of 200 μg of anti-IL-4 Ab (BioXCell) and 100 μg of anti-IL-21 Ab (R&D Systems) were performed for 2 days after T-cell transfer. Two hundred μg of anti-IL-10 Ab (JES5-2A5; BioXCell) was injected every other day after tumour inoculation. For in vivo cell depletion, mice were injected with 200 μg of anti-CD4 Ab (GK1.5), anti-CD8 Ab (2.43) or anti-Gr-1 Ab (R56–86; BioXCell) 4–5 days before T-cell transfer, immunization or tumour inoculation.
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