In this study we assessed the in vitro toxicity of 14 paired indoor and outdoor PM(2.5) samples (particulate matter < or =2.5 microm in aerodynamic diameter) collected in 9 Boston-area homes. Samples were collected as part of a large indoor particle characterization study that included the simultaneous measurement of indoor and outdoor PM(2.5), particle size distributions, and compositional data (e.g., elemental/organic carbon, endotoxin, etc.). Bioassays were conducted using rat alveolar macrophages (AMs), and tumor necrosis factor (TNF) was measured to assess particle-induced proinflammatory responses. Additional experiments were also conducted in which AMs were primed with lipopolysaccharides (LPS) to simulate preexisting pulmonary inflammation such as that which might exist in sick and elderly individuals. Significant TNF production above that of negative controls was observed for AMs exposed to either indoor or outdoor PM(2.5). TNF releases were further amplified for primed AMs, suggesting that preexisting inflammation can potentially exacerbate the toxicity of not only outdoor PM(2.5) (as shown by previous studies) but also indoor PM(2.5). In addition, indoor particle TNF production was found to be significantly higher than outdoor particle TNF production in unprimed AMs, both before and after normalization for endotoxin concentrations. Our results suggest that indoor-generated particles may be more bioactive than ambient particles. Endotoxin was demonstrated to mediate proinflammatory responses for both indoor and outdoor PM(2.5), but study findings suggest the presence of other proinflammatory components of fine particles, particularly for indoor-generated particles. Given these study findings and the fact that people spend 85-90% of their time indoors, future studies are needed to address the toxicity of indoor particles.