The levels of ultrafine particles (UFPs) and their size distributions on a highway electronic toll collection (ETC) lane were measured from October 30 to November 1 and November 5 to November 6, 2008. The hourly UFP levels measured at the highway ETC lane were 1.3 × 104–1.9 × 105 particles cm−3 (mean = 9.4 × 104 particles cm−3). Compared with urban UFP levels, average UFP levels at the highway ETC lane were about 5–10 times higher than those previously measured in urban areas, indicating that a considerable amount of UFPs were exhausted from vehicles. At the highway ETC lane, the average UFP number size distribution had a dominant mode at about 10 nm and a minor mode at about 33 nm. Measurement results indicate that nucleation mode particles remained at relatively high levels compared to Aitken mode and accumulation mode particles at the highway ETC lane. This study identifies the impacts of traffic volumes and wind speeds on ambient UFP levels. Measurement results show that the elevated UFP levels resulting from traffic volume decreased exponentially as wind speed increased on the highway when wind speed was <2.0 m s−1. However, the elevated UFP levels resulting from traffic volume increased slightly when wind speed was >2.0 m s−1 due to local turbulent mixing causes UFPs at high wind speeds. According to measurement results, high levels of UFP were observed at low wind speeds and under high traffic volumes.