While increasing numbers of cytomegalovirus (CMV)-associated diseases are occurring in patients undergoing conventional chemotherapy, information regarding CMV reactivation is limited. This pilot study was conducted to investigate CMV reactivation induced by chemotherapy. Seven blood samples were collected from each of 15 patients with newly diagnosed malignant disease, at baseline before chemotherapy, and once every month after chemotherapy was commenced. CMV viral loads in leukocytes were determined by real-time PCR. Host responses to changes in viral loads were assessed by assaying CMV-specific IgG titres and tumour necrosis factor (TNF)-alpha and interferon (IFN)-gamma levels in each of the blood samples, and by scoring the number of CMV-associated clinical symptoms that developed. All except one patient experienced CMV reactivation during the course of chemotherapy, with the average viral load peaking after the third course of treatment. Titres of CMV-specific IgG increased in line with the increase in viral load. Plasma levels of TNF-alpha and IFN-gamma initially decreased from baseline, and then rose to peak levels at the same time as, or shortly after, the highest viral loads were recorded. Clinical symptoms potentially attributable to CMV infection appeared as the viral load increased. It was concluded that the incidence of CMV reactivation in patients receiving conventional chemotherapy is high. Reactivation is not asymptomatic, but was self-limiting in most of these cases. Increases in plasma TNF-alpha and IFN-gamma occur after reactivation, but not before.