The present study aimed to establish whether 2 weeks of high-intensity interval training would have a beneficial effect on aerobic fitness, fat oxidation, blood pressure and body mass index (BMI) in healthy adolescent boys. Ten adolescent boys (15.1 ± 0.3 years, 1.3 ± 0.2 years post-estimated peak height velocity) completed six sessions of Wingate-style high-intensity interval training over a 2-week period. The first session consisted of four sprints with training progressed to seven sprints in the final session. High-intensity interval training had a beneficial effect on maximal O2 uptake (mean change, ±90% confidence intervals: 0.19 L · min(-1), ±0.19, respectively), on the O2 uptake at the gas exchange threshold (0.09 L · min(-1), ±0.13) and on the O2 cost of sub-maximal exercise (-0.04 L · min(-1), ±0.04). A beneficial effect on the contribution of lipid (0.06 g · min(-1), ±0.06) and carbohydrate (-0.23 g · min(-1), ±0.14) oxidation was observed during sub-maximal exercise, but not for the maximal rate of fat oxidation (0.04 g · min(-1), ±0.08). Systolic blood pressure (1 mmHg, ±4) and BMI (0.1 kg · m2, ±0.1) were not altered following training. These data demonstrate that meaningful changes in health outcomes are possible in healthy adolescent boys after just six sessions of high-intensity interval training over a 2-week period.