Comparison of extracellular adenosine, CGS21680 and anti-CD3 mAb-induced apoptosis and cell death in mouse thymocytes.
Ex vivo thymocytes were incubated 16 hours at 37°C in 5% CO2 incubator in the presence of adenosine or adenosine analog CGS21680 or anti-CD3 mAb, and the extent of apoptosis or cell death was measured by flow cytometry as described in “Materials and methods.” (A) Detection of apoptotic events in thymocytes using Annexin V assay; (B) comparable proportion of thymocytes is lost after extAdo-induced and TCR/CD3-induced apoptosis. Cell loss was estimated by flow cytometry by PI staining of dead cells. (C) Demonstration of typical experimental gate selection for evaluation of number and proportion of live (gate R1), apoptotic (gate R2), and dead (gate R3) after 16 hours of incubation of thymocytes in vitro with CGS21680 as panel A. The status of cells was studied by flow cytometry after their staining with propidium iodide (PI) and Annexin V (upper graph) and by their site versus forward scatter (lower graph).
