Bcl-2 overexpression in cardiac allografts results in less cardiomyocyte apoptosis, immune cell infiltration, and production of T_H1 cytokines, chemokines, and adhesion molecule 30 days after transplantation. Thirty days after transplantation, cardiac allografts were harvested, and histologic sections and lysates from the cardiac allografts were prepared, as described in “Materials and methods.” (A) The number of apoptotic cells in cardiac allografts was counted by ISOL TUNEL staining, and the activity of caspase-3 and the level of Fas ligand in heart lysates were measured using ELISA. (B) Histologic sections were also stained with anti-CD3, -CD4, -CD8a, -CD11c, -Mac-1, and -B220 antibodies, as described in “Materials and methods.” The number of positive cells in each cardiac allograft was manually counted by 2 investigators blinded to the experimental conditions. Cells were counted in 8 animals (4 fields each) at 200 × magnification. The percentage of positively stained cells—that is, the number of labeled cells divided by total number of cells—was recorded. (C) Production of TNF-α, IFN-γ, MCP-1/CCL2, IP-10/CXCL10, MIG/CXCL9, ICAM-1, and VCAM-1 in lysates from the cardiac allografts was measured by ELISA. Filled columns show the data from WT heart grafts (n = 8), and open columns show data from the Bcl-2 Tg heart grafts (n = 8). Mean ± SD values are shown.