This study aimed to clarify the effects of uncomfortable thermal environments within clothing on brain activation, autonomic nervous system activity, and work efficiency. Oxy-Hb concentrations in the prefrontal cortex were measured using near-infrared spectroscopy (NIRS), and autonomic nervous system activity was analyzed using electrocardiography. The participants were twelve healthy women in their 20s, but the valid number of participants was nine. In a laboratory at 28°C and 50% RH, the environment within participants’clothing was changed using a moisture-impermeable raincoat. Participants performed a calculation task as a stress task under comfortable and uncomfortable conditions, with the latter condition involving high temperature and humidity within clothing. The results showed that work efficiency did not improve in the uncomfortable garment environment despite increasedΔoxy-Hb concentrations of a calculation task minus a vocalization task. In contrast, various autonomic nervous system indices derived from Lorenz plots using RRI showed that when the clothing environment was uncomfortable, the sympathetic nervous system tended to become dominant, and the parasympathetic nervous system tended to become subordinate, thereby increasing stress. In conclusion, it was found that an increase inΔoxy-Hb under stressful conditions caused by an uncomfortable clothing environment did not lead to improved work efficiency.