Living in deprived neighbourhoods may have biological consequences, but few studies have assessed this empirically. We examined the association between neighbourhood deprivation and allostatic load, a biological marker of wear and tear, taking into account individual’s socioeconomic position. We analysed data from three cohort studies (CoLaus-Switzerland; EPIPorto-Portugal; Whitehall IIUK) comprising 16,364 participants. We defined allostatic load using ten biomarkers of dysregulated metabolic, cardiovascular, and inflammatory systems (body mass index; waist circumference; total, high and low density lipoprotein cholesterol; triglycerides; glucose; systolic and diastolic blood pressure; C-reactive protein). Mixed Poisson regression models were fitted to examine associations with neighbourhood deprivation (in quintiles, Q1-least deprived as reference). After adjustment for confounding variables, participants living in the most deprived quintile had 1.13 times higher allostatic load than those living in the least deprived quintile (Relative Risk, RR, for Q2 RR = 1.06, 95% CI 1.03- 1.09; Q3 = 1.06, 1.03–1.10; Q4 = 1.09, 1.06–1.12; Q5 = 1.13, 1.09–1.16). This association was partially modified by individual’s socioeconomic position, such that the relative risk was higher in participants with low socioeconomic position (Q5 vs Q1 1.16, 1.11–1.22) than those with high socioeconomic position (Q5 vs Q1 1.07, 1.01–1.13). Neighbourhood deprivation is associated with biological wear and tear, suggesting that neighbourhood-level interventions may yield health gains.