Opiates and opioid agents are known to affect the immune system. In humans this includes alterations in natural killer (NK) cell activity. Morphine is reported to reduce in vivo spleen NK activity in rats, whereas for methadone only in vitro data have been described. In the present paper we describe a systematic study on the chronic effects of well-known opiates, comparing for the first time the effects of morphine and methadone on NK cell activity in various organs: in addition to spleen, also in the peritoneal cavity, and lungs. In all organs the NK activity was determined using three effector: target cell ratios. Morphine and methadone given by food during 6 weeks decreased the NK cell activity in rat spleen, supporting published data on morphine. The role of the opiate receptor is discussed. However, the overall action of morphine could not be described as suppressive because stimulation of NK cell activity in the peritoneal cavity and lungs by morphine was found. In contrast, methadone induced a decrease in the NK cell activity in these organs. Apart from these differential expressions of morphine- and methadone associated effects on NK cell activity, the findings demonstrate the potential adverse effects of these opiates on an important antiviral defence mechanism.