Body weight supported treadmill training assisted with a robotic driven gait orthosis is an emerging clinical tool helpful to restore gait in individuals with loss of motor skills. However, the autonomic response during this rehabilitation protocol is not known. The aim of the study was to evaluate the autonomic response during a routine protocol of motor rehabilitation through spectral and symbolic analyses of short-term heart rate variability in a group of 20 healthy subjects (11 men, mean age 25+/-3.8 years). The protocol included the following phases: (1) sitting position; (2) standing position; (3) suspension during subject instrumentation; (4 and 5) robotic-assisted treadmill locomotion at 1.5km/h and 2.5km/h respectively with partial body weight support; (6) standing recovery after exercise. Results showed a significant tachycardia associated with the reduction in variance during the suspended phase of the protocol compared to the sitting position. Spectral analysis did not demonstrate any significant autonomic response during the entire protocol, while symbolic analysis detected an increase in sympathetic modulation during body suspension and an increase of vagal modulation during walking. These results could be used to improve understanding of the cardiovascular effects of rehabilitation in subjects undergoing robotic driven gait orthosis treadmill training.