Epidemiologic studies indicate that cigarette smoking (CS) increases the risk and severity of acute lung injury (ALI)/acute respiratory distress syndrome (ARDS). The mechanism is not understood at least in part due to lack of animal models that reproduce the key features of CS priming process. In this study, using two strains of mice we characterized a double-hit mouse model of ALI induced by CS priming of injury caused by lipopolysaccharide (LPS). C57BL/6 and AKR mice were pre-exposed to CS briefly (3 hours) or sub-acutely (3 weeks) before intra-tracheal instillation of LPS and ALI was assessed 18 hours after LPS administration by measuring lung static compliance, lung edema, vascular permeability, inflammation and alveolar apoptosis. We found that as little as 3 hours of exposure to CS enhanced LPS-induced ALI in both strains of mice. Similar exacerbating results were observed after 3 weeks of pre-exposure to CS. However, there is a strain difference in susceptibility to CS priming for ALI, with a greater effect in AKR mice. The key features we observed suggest that 3 weeks of CS pre-exposure of AKR mice is a reproducible, clinically relevant animal model that is useful for studying mechanisms and treatment of CS priming for a second hit-induced ALI. Our data also support the concept that increased susceptibility to ALI/ARDS is an important adverse health consequence of CS exposure that needs to be taken into consideration when treating critically ill individuals.