Figure 6
Effect of IL-6 on Mage-b-induced immune responses in vitro. To analyse the effect of IL-6 on Mage-b-specific immune responses in vitro, draining lymph nodes (LNs) of 4T1 tumour-bearing mice were cocultured with or without 64pT tumour cells, expressing highly Mage-b, and producing high levels of IL-6. These cocultures were performed in the absence or presence of anti-IL-6 antibodies (A). After 2 days of stimulation, the production of IFNγ was determined with quantitative ELISA. In this experiment, the lymph nodes (LNs) of 10 mice were pooled. In addition, spleen cells of 4T1 tumour-bearing mice were cocultured with or without autologous bone marrow (BM) cells transfected with pcDNA3.1-Mage-b and pCMV-GM-CSF. These cocultures were performed in the absence or presence of purified IL-6 (B). After 2 days of stimulation, the number of IFNγ-producing cells was determined with an ELISPOT reader. This experiment was performed twice with spleens of mice that received vaccination with LM-LLO-Mage-b/2nd. In each experiment, spleens of five mice were pooled. Controls such as BM cells transfected with pcDNA3.1-Mage-b and/or pCMV-GM-CSF, or non-transfected BM cells, did not produce IFNγ (data not shown). The results of both assays were subjected to statistical analysis using the unpaired t-test. The error bars represent the s.e.m.
