Derived from inclusions late in the infectious cycle induce aponecrosis in human aortic endothelial cells-1

D (D). were detected with anti -MOMP (green), DNA strand breaks with TUNEL (red), cytoplasm with NHS-biotin (yellow) and DNA of bacteria and host cells with DAPI (blue). Cells in random fields were counted (n = 200) and the percentage of NHS-biotin-TUNEL positive cells was calculated (G, H). A: HAEC infected with 20 IFU/cell 48 hpi displaying normal nuclear morphology and two MOMP positive inclusions (asterisks). B: HAEC infected with 20 IFU/cell 48 hpi that show a spot-like infection and TUNEL positive condensed nuclei as well as cytoplasmic NHS-biotin signal. -MOMP positive spots and aggregates (arrows) are randomly scattered in the cell. C: Chloramphenicol treated cells infected with 20 IFU/cell 48 hpi lack TUNEL and NHS-biotin labelling and display a regular nuclear morphology. D: Uninfected HAEC displaying a regular nuclear structure without TUNEL and cytoplasmic NHS-biotin label. E: HAEC infected for 24 h with 40 IFU/cell. F: HAEC infected for 72 h with 5 IFU/cell. Note that although the cells are infected with different infection doses and fixed at different time points they both elicit the same cell death morphology, thus excluding a cytotoxic effect and underlining the independency of cell death morphology from infection time. Scale bar; 10 μm G (left panel): The number of double positive cells (NHS-biotin + TUNEL) increases dose dependently. The number of single NHS-biotin positive cells does not significantly vary among different infection doses and time points. G (right panel): Chloramphenicol treated infected cells display a lower amount of NHS-biotin-TUNEL positive cells which is independent of the initial infectious dose compared to untreated cells. H: The number of NHS-biotin-TUNEL positive cells increases time dependently in HAEC infected with low infection doses whereas in cells infected with high concentrations the number of NHS-biotin-TUNEL positive cells decreases over time. One representative experiment out of three is shown.<p><b>Copyright information:</b></p><p>Taken from "derived from inclusions late in the infectious cycle induce aponecrosis in human aortic endothelial cells"</p><p>http://www.biomedcentral.com/1471-2180/8/32</p><p>BMC Microbiology 2008;8():32-32.</p><p>Published online 19 Feb 2008</p><p>PMCID:PMC2275732.</p><p>