AbstractWe aimed to investigate whether health-related quality of life (HRQoL) is associated with retinal vascular caliber, an indicator of subclinical cardiovascular disease risk. 1600 students aged between 11–19 years (821 girls and 779 boys) were examined during 2009–2011. Retinal vessel caliber was measured from digital retinal images. HRQoL was assessed by the Pediatric Quality of Life Inventory (PedsQL). In the overall cohort, each 1-unit increase in PedsQL total score and the psychosocial summary score was associated with ~0.05 μm narrowing in retinal arteriolar caliber (multivariable-adjusted p-value = 0.01). Participants in the lowest versus highest tertile of PedsQL total score, psychosocial summary, social and school item scores had significantly wider retinal arteriolar caliber: 161.7 μm versus 160.2 μm (p = 0.02); 161.6 μm versus 160.0 μm (p = 0.02); 161.6 μm versus 159.9 μm (p = 0.002); and 161.6 μm versus 159.9 μm (p = 0.01), respectively. Significant interactions (p < 0.05) were observed between gender and PedsQL total score with retinal arteriolar calibre. In boys, inverse associations were observed between PedsQL total score (p = 0.01), psychosocial summary (p = 0.01), and social scores (p = 0.01) and retinal arteriolar caliber. No significant associations were observed between PedsQL scores and retinal vessel caliber in girls. Diminished HRQoL in adolescents was independently associated with structural retinal microvascular changes.