The aim of this study was to investigate the effect of long-term consumption of caffeine in the development of Ehrlich ascites carcinoma (EAC) cells in adult female mice, 25-30 g, in relation to immune response. Mice were treated with caffeine (20 mgkg-1 daily, p.o.) for 22-27 consecutive days or inoculated with EAC cells (5 × 106 cells/mL, i.p.), or both. Control mice, corresponding to experimental groups, were treated with corresponding vehicles under similar conditions. The lymphocyte viability, mitogen-induced proliferating activity, cytotoxicity and DNA fragmentation from blood, spleen and thymus of both control and experimental groups were measured as immune response parameters. An immune response index, corticosterone, was also measured in adrenals and plasma under similar conditions. Results showed that development of EAC cells caused immune suppression with a reduction of lymphocyte viability, cytotoxicity and proliferative activity and induction of DNA fragmentation in those tissues, as well as an increase in plasma corticosterone. Though long-term caffeine treatment (which resulted in tolerance to caffeine) alone did not alter significantly any of the immune response parameters studied, including corticosterone status (immune biomarker), the continuation of caffeine treatment during the development of EAC cells either restored or reduced the EAC cell-induced alteration in these parameters, including the HPA axis biomarker. These results suggest that long-term caffeine intake may inhibit or reverse the EAC cell-induced immune suppression. © 2007 The Authors.