A cross-sectional survey was carried out to evaluate the relation between occupational exposure to low levels of anaesthetic gases (nitrous oxide and isoflurane) and immune parameters. Fifty-one anaesthetists were recruited among different Services of Anaesthesiology and Reanimation. The control group consisted of non-exposed physicians, similar for gender, age, and job grade. Total number of lymphocytes, lymphocyte subpopulations and the natural killer (NK) cytotoxic activity were measured. Information on personal and professional characteristics and on short- and long-term exposure was collected. Percentages of T cells (CD3) decreased significantly in anaesthetists compared to controls, whereas numbers of NK cells (CD16+ CD3-) increased. After correction for confounders, short-term (last 2 weeks) exposure was associated with a decrease in percentages of total T and T helper (CD4) cells. Furthermore, T helper percentages were significantly reduced with increasing individual exposure score evaluated on the basis of working days and levels of anaesthetic gases in operating rooms. A significant X-ray-associated increase of numbers and percentages of NK cells was lastly observed. Despite limited present exposure to anaesthetic gases, a specific derangement in lymphocyte subpopulations, with T lymphocytes more affected than B, has been observed.