Figure 5

GIM can mediate non-MHC-restricted tumor cytotoxicity. GIM, peripheral blood mononuclear cells, and normal (5 × 10⁶, 1 × 10⁶, or 5 × 10⁵) microglia were each incubated as an effector population with 1 × 10⁵ carboxy-fluorescein diacetate succinimidyl ester-labeled U-87 target cells, for an effector-to-target ratio of 50:1, 10:1, or 5:1, respectively. Propidium iodide was added after 24 hours incubation, and cells were analyzed by flow cytometry. Cells were first gated only on the carboxy-fluorescein diacetate succinimidyl ester⁺ target population (excluding all other cells in the assay), and the propidium iodide expression on these cells was determined. The percentages of cytolytic activity of the effector cells were calculated (Y axis)(dead targets in upper right quadrant/[dead targets in upper right quadrant + live targets in lower right quadrant]) and plotted against each respective effector-to-target ratio (X axis). These data are from one tumor specimen and are representative of experiments from cells isolated from 3 different glioma patients. Bars at each data point represent 95% confidence intervals for each proportion. After 24 hours, GIM were functional in their tumor cytotoxic activity and comparable to PBMCs, however cytotoxic activity of GIM was significantly (*p < 0.0001) lower than that of normal microglia.