Automatic adjustments of muscle activity throughout the body are required for the maintenance of balance during human walking. One mechanism that is likely to contribute to this control is the heteronymous spinal excitation between human ankle dorsiflexors and knee extensors (CPQ-reflex). Here, we investigated the CPQ-reflex at different walking speeds (1-6 km/h) and stride frequencies (0.6-1.3 Hz) in healthy human subjects to provide further evidence of its modulation, and its role in ensuring postural stability during walking. The CPQ-reflex was small or absent at walking speeds below 2-3 km/h, then increased with walking speeds about 3-4 km/h, and reached a plateau without any further change at walking speeds from 4 to 6 km/h. The reflex showed no modulation when the stride cycle was varied at constant speed (4 km/h; short steps versus long steps). These changes were unlikely to be only caused by changes in the background EMG activity and modifications in peripheral input, and likely reflected central modulation of transmission in the involved reflex pathways as well. It is suggested that the purpose of the reflex is to ensure knee stability at moderate-to-high walking speeds.