As gender may influence thermal and air quality perception in indoor environments, the aim of this study was to analyse gender influence on air movement, air humidity, air quality and thermal perception in office buildings in Southern Brazil. Statistical descriptions, regression analyses and hypothesis tests were performed using data collected from field studies conducted in a fully air-conditioned building and in three mixed-mode buildings. In addition, comfort temperatures were estimated through the Griffiths method. Results showed that females tend to feel colder compared to males. Men and women tended to present higher thermal acceptability and thermal comfort in mixed-mode buildings and in fully air-conditioned buildings, respectively. Weak but significant correlations were obtained between some environmental and subjective variables. In general, comfort temperatures were statistically higher for females (24.2 °C) than for males (23.5 °C). Significant gender differences for thermal perceptions of indoor environments were detected.