Objectives: In Poland, occupational exposure to cold microclimate is quite common (5.1 workers/1000 occupationally active people). Reports on health effects of this exposure are rather scarce. The aim of the study was to evaluate the physiological reaction in workers occupationally exposed to cold microclimate. Materials and Methods: Examinations were performed in a group of 102 workers (41 women and 61 men) employed at cold storage units. The mean age in the group was 39.1 ± 9.9 years and the duration of employment under conditions of cold environment was over 12 years. The study population was divided into four groups, according to microclimate conditions (group I, ambient temperature 26°C; group II, 10-14°C; group III, 18-20°C, control group; and group IV, 0-10°C). The workers underwent the following procedures: general medical examinations, cold pressor test, ambulatory blood pressure monitoring, and heart rate variability (HRV) analysis (time- and frequency-domain parameters). Results: The results were adjusted for confounding factors (age, smoking and drinking habits). The analysis of HRV parameters did not reveal any significant differences between the study groups. However, systolic and diastolic blood pressure (BP) in the daytime and at night was significantly higher in group IV compared to group II. Mean heart rate (HR) in the daytime and at night and the BP and HR day/night ratio did not differ between the groups. The analysis of BP by gender revealed that in women, systolic BP during the day and at night was significantly higher in group IV than in group II. In the group of workers with hypertension (18 men and 5 women), men reacted to the cold pressor test either by increased or decreased BP while all the women reacted by the increased BP. Conclusions: Our findings indicated that in workers exposed to cold microclimate, the physiological reaction was dependent on gender and ambient temperature. Women seemed to be more sensitive to cold stress than men. However, this finding must be further investigated.