Objective: Recombinant human interferon alpha (rhIFN-a) is used therapeutically in malignant disorders and chronic hepatitis. The present study was assessed to study the effects of rhIFN-a on the hypothalamic±pituitary±testicular (HPT) axis. Design and methods: We performed a saline-controlled cross-over study in six healthy men, sequentially measuring the serum concentrations of gonadotropins, testosterone, the free androgen index (FAI) and sex hormone-binding globulin (SHBG) after a bolus subcutaneous injection of rhIFN-a. Results: rhIFN-a induced a sustained decrease of both testosterone (from 19.5 6 1.88 to a nadir of 5.49 6 0.51 nmol/l at the end of the study) and FAI (from 98.7 6 14.7 to a nadir of 32.1 6 5.3 at the end of the study), whereas concentrations of LH, FSH and SHBG were not different between the two studies. Conclusions: Our results suggest that rhIFN-a affects the HPT axis at the testicular level, either directly or indirectly, and changes feedback relationships between the pituitary and the testis.