Single-cell suspensions of mouse lung CD11bϩ and CD11bϪ cells were prepared and plated in 24-well plates at a concentration of 10 6 cells in 1 ml of complete RPMI-1640 for 1 hour. The supernatants were removed, and 2 ϫ 10 5 d7EB cells in fresh medium were added either directly with the mouse cells or on the insert membrane of the Transwell system (HTS Transwell 0.4-m pore size polycarbonate membrane; Corning, Inc.). As a control, CD11bϩ cells were plated without d7EB cells. The co-cultured cells were or were not stimulated with 1 g/ml of lipopolysaccharide (LPS) for 6 hours. At the end of the stimulation period, the supernatants were collected, and mouse-specific ELISAs (R&D Systems, Inc.) were performed on samples to detect the released mouse TNF-␣, IFN-␥, and IL-10. This protocol was repeated for the experiments with ACE and KDR sorted d7EB cells.