The purpose of this study was to determine age-related changes in lung lining fluid antioxidants in broilers reared under thermoneutral or cold temperature conditions. Male broilers (Cobb 500) were placed in floor pens within environmental chambers and fed a standard commercial starter diet. The thermoneutral Control chamber was maintained at 32, 30, 27, and 22 to 25 C for Weeks 1, 2, 3, and 4 to 7, respectively, whereas temperature in the Cold chamber was lowered to 18 C during Week 3 and maintained between 15 and 18 C for the rest of the study. At 2, 4, and 7 wk, four to six birds per chamber were selected randomly. The lungs were lavaged with heparinized saline (2 mL/g lung) to obtain lung lining fluid. Antioxidants [reduced (GSH), oxidized (GSSG), and total (TGSH) glutathione, uric acid, ascorbic acid, and alpha- and gamma-tocopherol] in lung lining fluid were determined by HPLC; protein was determined colorimetrically. In Controls, levels of alpha- and -gamma-tocopherol, uric acid, and GSH in lung lining fluid decreased between 2 and 7 wk of age. Birds in the Cold chamber exhibited higher protein, a higher GSSG:TGSH ratio, and a decrease in ascorbic acid (7 wk) in lung lining fluid relative to Controls. Lung lining fluid antioxidants were not correlated with antioxidants in plasma. To determine the effect of vitamin E supplementation on lung lining fluid antioxidants, birds were given a supplement of 200 IU alpha-tocopherol per day for 7 d. Alpha-tocopherol supplementation elevated alpha-tocopherol levels in lung lining fluid, but lowered ascorbic acid, GSH, and GSSG and had no effect on uric acid in lung lining fluid. The results of this study suggest that antioxidant protection in lung lining fluid may diminish with age, that cold conditions in this study produced an oxidative stress in lung lining fluid in broilers, and that oral supplementation of alpha-tocopherol elevated lung lining fluid alpha-tocopherol.