This study examined thermo-physiological responses according to the design change of construction site working clothes (control (C) working clothes; prototype (P) working clothes). We measured rectal temperature, skin temperature, micro-climate within the clothes and sweat rate. In the evaluation of physiological functionality, based on pattern improvement in working clothes, P working clothes showed significantly lower rectal temperatures, trunk and thigh skin temperatures than C working clothes. It is preferable that rectal temperature should be kept low during work that is not favorable to an increase in body temperature. P working clothes were more physiologically functional than C working clothes. In addition, P working clothes showed significantly lower temperatures in the trunk and thigh parts in a micro climate temperature. We could explain that the side seam zipper on the pants and the gusset on armpit parts create an air permeability effect of lowering the temperature of micro-climate. Aggressive ventilation through the slit of the garment is an important factor for the restoration of the physiological function of the worker at rest between work. Sweat rate showed a higher level in C working clothes than P working clothes. When working in a hot environment, workwear needs to be designed so that the worker is not exposed to thermal stress. Therefore, it was evaluated that the P work clothes used in this study alleviated the physiological burdens of heat.