Research-based evidence supports the effectiveness of soothing music in improving stress-related psycho-physiological indices in a clinical setting. However, there is currently insufficient scientific knowledge of the effects of music on immune markers of stress in humans. Therefore, the aims of the study were to compare the effects of music and quiet rest on the levels of interleukin-6 (IL-6), tumour necrosis factor-α (TNF-α), interleukin-10 (IL-10), heart rate and mean arterial pressure among healthcare workers. By using a randomized controlled trial design, 60 nurses were randomly assigned to the stimulating or sedating music or rest groups for 30 min. Participants' psychoneuroimmunological parameters were measured using enzyme-linked immunosorbent assays. General estimating equation was used to analyse data. Results revealed that IL-6, TNF-α and IL-10 were not detectable in this population. No significance differences in heart rate were found among the three groups. However, the stimulating music group had significantly higher mean arterial pressure levels than the sedating music group but no differences between the quiet rest group and the sedating music group. Music with different tempi had little effect on mean arterial pressure. Any effect of music on immune markers of stress requires further research.