The thermal comfort of a work glove affects wear compliance. This study investigates the thermal comfort of five types of anti-vibration gloves that use chloroprene rubber and spacer fabric as materials to isolate vibration. An evaluation of the fabric and material on the palm and dorsal of the glove, and a wear trial of the gloves were subsequently carried out. The results showed that the palm of the glove made of spacer fabric had higher air and water vapour permeabilities than chloroprene rubber. Using a thin mesh fabric for the dorsal of the glove resulted in a significantly lower skin temperature than using spacer fabric or chloroprene rubber during hand activity. The thermal sensation of the subjects showed no significant difference among the five glove samples. However, the glove wear comfort of the spacer fabric glove was the lowest. Recommendations for the fabric thickness and glove design are provided for developing anti-vibration gloves.