People in polluted communities are often exposed to both PM and ozone (O3), albeit not always simultaneously; an important question is whether exposure to particles with seasonal compositional differences can influence biological outcomes. We addressed this question using a mouse model of cardiovascular disease by contrasting the health outcomes of exposures to particles formed or aged during periods of relatively high photochemical activity (i.e. spring/summer), which has increased ambient O3 concentrations, with outcomes of exposures to fall/winter particles which are associated with lower O3 concentrations. Electrocardiographs (ECGs) and blood pressures (BPs) were acquired following exposures to concentrated ambient particles (CAPs). ECGs were analyzed to changes in specific waveform parameters and changes in heart rate variability (HRV). Exposures elicited several types of waveform abnormalities that were associated with seasonal differences in particle constituents. Alterations in R-R interval and P-R interval were seen following exposure to summer CAPs but not fall CAPs and differential responses were seen in the corrected Q-T interval following the two seasonal exposures. Measures of HRV increased after exposure to summer CAPs compared to air-exposed controls but not following the winter CAPs exposure. There were chemical differences with respect to the organic constituents in ambient particles between summer and fall aerosol. The oxygen to carbon ratios (O:C) were generally higher in the spring and summer than in the fall, consistent with seasonal differences in atmospheric photochemical activity. Seasonal differences in atmospheric photochemical activity can modify ambient aerosol composition and can alter biological responses in the cardiovascular system. The results from this study confirm that ambient photochemical activity can alter the toxicity of ambient PM. Regional and seasonal differences in PM2.5 composition should be important considerations when evaluating the effects of PM exposure on cardiovascular health.Implications: Particles formed during periods of high photochemical activity (e.g. spring/summer) elicit more adverse cardiovascular health effects than particles formed during periods of low photochemical activity (e.g. fall/winter). Seasonal differences in atmospheric photochemical activity modified ambient aerosol composition and worsened cardiovascular responses. These results can inform regulatory agencies and may help design air quality regulations for PM2.5 that consider seasonal and regional variations.