Diminished in vitro proliferation and altered mitotic profiles of AD DC lymphocytes. Lymphocytes collected from various AD DC subjects (UPN) and age-matched controls were placed in long-term culture supplemented with soluble anti-CD3 (1 μ g/mL), anti-CD28 (0.5 μ g/mL), and IL-2 (50 U/mL). For mitoses determinations, cells were loaded with CFSE as per manufacturers instructions. (A) Proliferation was quantitated by counting live cells and is expressed as percent fold difference in total cell number, over time, relative to healthy age-matched controls (unique patients represented by different symbols). P values (as calculated by comparing mean value relative to control to determine if significantly different than 1 at each time point) were significant at days 6 to 7 (P = .008) and days 8 to 9 (P = .0006). (B) Example of gating strategy in FL1 wavelength for determining number of mitoses (M = 0, 1-2, or ≥ 3). (C) Mitotic profile for 7 AD DC subjects, expressed as a percentage of gated cells within the entire cell population undergoing varying numbers (0, 1-2, or ≥ 3) of mitoses on day 4 in culture, relative to control values. The P values (as calculated from the average percent of cells from the patients versus the controls in each mitotic grouping to determine if mean value relative to control was significantly different from 1) are displayed in the inserted box. Bold line on y axis represents control value.