Rationale: Lung injury after cigarette smoking is related to particle retention. Iron accumulates with the deposition of these particles. Objectives: We tested the postulate that 1) injury after smoking correlates with exposure to the particulate fraction of cigarette smoke, 2) these particles alter iron homeostasis triggering metal accumulation, and 3) this alteration in iron homeostasis affects oxidative stress and inflammation. Methods: Rats and human respiratory epithelial cells were exposed to cigarette smoke, filtered cigarette smoke, and cigarette smoke condensate (CSC; the particulate fraction of smoke) and indices of iron homeostasis, oxidative stress, and inflammatory injury were determined. Comparable measures were also evaluated in non-smokers and smokers. Measurements and Main Results: After exposure of rats to cigarette smoke, increased lavage concentrations of iron and ferritin, serum ferritin levels, and non-heme iron concentrations in the lung and liver tissue all increased. Lavage ascorbate concentrations were decreased supporting an oxidative stress. After filtering of the cigarette smoke to remove particles, most of these changes reversed. Exposure of cultured respiratory epithelial cells to CSC caused a similar accumulation of iron, metal-dependent oxidative stress, and increased interleukin-8 release. Lavage samples in healthy smokers and smoking patients with chronic obstructive pulmonary disease revealed elevated concentrations of both iron and ferritin relative to healthy non-smokers. Lavage ascorbate decreased with cigarette smoking. Serum iron and ferritin levels among smokers were increased supporting systemic accumulation of this metal after cigarette smoke exposure. Conclusions: We conclude that cigarette smoke particles alter iron homeostasis both in the lung and systemically.