Previous work demonstrated that splenic natural killer (NK) cell cytolytic activity was suppressed in ethanol-consuming mice within 1 week and for as long as 10 weeks concurrent with ethanol intake. However, it is unknown if suppression of NK cell activity with ethanol consumption results from regional losses in NK cells. The present experiments were designed to examine the effects of ethanol on the percentage and total number of NK cells in the spleen and blood. Data indicate that, after 4 weeks of ethanol intake, NK cell activity of peripheral blood lymphocytes was also suppressed (50% of controls). With short-term (2-week) exposure, the percentage of NK cells in the blood and the spleen remained relatively constant. In the spleen, changes in the number of NK cells reflected generalized fluctuations in lymphocyte numbers rather than variation in the percentage of NK cells. NK cells were most sensitive to the effects of ethanol during long-term (8-week) ethanol intake, as demonstrated by significant reductions in the total percentage of NK1.1+ cells in the blood, and the LGL-1+ subset in the blood and spleen. These results provide insight into the mechanism of NK modulation by ethanol. Because NK cells were not selectively depleted after 2-4 weeks of ethanol intake, loss of cytolytically active NK cells cannot explain the suppression of in vitro cytolytic activity of cells from ethanol-consuming mice. However, with prolonged ethanol intake (8 weeks), the depletion of NK cells, specifically the LGL-1+ subset, may contribute to the overall suppression of NK cell cytolytic function.