Abstract Background Tramadol hydrochloride is a centrally acting analgesic drug that has turned out to be a drug of growing interest due to its success in the management of pain in humans. By considering the twin mechanism of action of tramadol, as the activation of the opioid and of the monoaminergic systems, the potential immunological effects of this drug should be considered and be evaluated. Studies show that Tramadol induces an improvement of postoperative immune suppression and increases NK cell activity, lymphocyte proliferation, and IL-2 production which may be linked to its addictive potentiality. The present study aims to investigate the possible association between opioid use disorder (tramadol) and the serum level of interleukin-2 (IL-2). An observational cross sectional study conducted at the Outpatient Clinic and Inpatient Unit of the Addiction Management Unit of the Neurology and Psychiatry Department of Assiut University Hospital. Assessment of serum level of IL-2 for 59 male patients diagnosed with opioid use disorder (tramadol) according to DSM-5 compared to 32 cross-matched male individuals as a control group. Results Comparing the serum level of IL-2 in patients with opioid use disorder (tramadol) with that of the controls, it was found that the patients with opioid use disorder (tramadol) have significantly higher serum level of IL-2 (318.69 ± 35.89) than that of controls (260.45 ± 30.08) (P = 0.01). Patients with tramadol urine assay (> 200 ng/ml) have significantly higher serum level of IL-2. Conclusion The current study found that patients with opioid use disorder (tramadol) have significantly elevated serum levels of IL-2 than healthy controls. Also, this elevation was dose related.