The toxicity of two important chemical components of fine ambient particulate matter (PM 2.5)-ammonium bisulfate (ABS) and elemental carbon (C)-was studied using aged (senescent) rats. The study tested the hypotheses that fine particle exposure can damage lungs and impair host defenses in aged rats and that ozone would potentiate the toxicity of these particles. Ammonium bisulfate aerosols were generated by nebulization of dilute aqueous solutions. Elemental carbon was generated from an aqueous suspension of carbon black. Carbon and ABS mixtures were generated by nebulization of a suspension of carbon black in a dilute aqueous solution of ABS. Rats were exposed, nose-only, for 4 h a day, three consecutive days a week, for 4 weeks. The rats were exposed to one of six atmospheres: (1) purified air; (2) C, 50 microg m(-3), 0.3 microm mass median aerodynamic diameter (MMAD); (3) ABS, 70 microg m(-3), 0.3 microm MMAD; (4) O3, 0.2 ppm; (5) ABS + C, 0.46 microm MMAD; and (6) ABS + C + O3, 0.45 microm MMAD. Data were analyzed using ANOVA and Tukey multiple comparison tests; a two-tailed significance level of 0.05 was used. The nuclei of lung epithelial and interstitial cells were examined to determine the labeling of the DNA of dividing cells by 5-bromo-2-deoxyuridine and to identify the location of injury-repair-related cell replication. Increased labeling of both epithelial and interstitial lung cells occurred following all pollutant exposures. Although epithelial cells are most likely impacted by inhaled particles first, the adjacent interstitial cells were the cells that showed the greatest degree of response. Exposure to the ABS + C + O3 mixture resulted in losses of lung collagen and increases in macrophage respiratory burst and phagocytic activities that were statistically significant. Our results demonstrate that ozone can increase the toxicity of inhaled particles (or vice versa), and suggest that detailed study of mixtures could provide a more comprehensive understanding of the mechanisms by which inhaled pollutants adversely affect human health.