Perforin enhances the granulysin-induced lysis of Listeria innocua in human dendritic cells-3

<p><b>Copyright information:</b></p><p>Taken from "Perforin enhances the granulysin-induced lysis of Listeria innocua in human dendritic cells"</p><p>http://www.biomedcentral.com/1471-2172/8/14</p><p>BMC Immunology 2007;8():14-14.</p><p>Published online 16 Aug 2007</p><p>PMCID:PMC1976101.</p><p></p> perforin in a concentration of 2.5 kU/ml. At indicated time points fluorescent images (Excitation 340 nm; Emission 525 nm) were recorded (A). The excitation ratio 340/380 nm was monitored over the whole experimental period (B). Some cultures were pretreated with TMB-8 (10 μM) and dantrolene (25 μM) to block intracellular Camobilization (C). (D), DC were equilibrated in a low Ca(1.3 μM) buffer before a first exposure to perforin, after washout and buffer exchange to normal Caconditions (1.3 mM) the cells were again treated with perforin. ATP and ionomycin were used as positive controls for Camobilization from intracellular or extracellular sources. One representative measurement of more than six independent experiments is presented. (E), -challenged DC were incubated for 3 hours with granulysin in presence or absence of 2.5 kU/ml perforin or 1 μM ionomycin at 37°C or were incubated with ionomycin alone. After the incubation, the cells were lysed in ice-cold water and viability was calculated from bacterial growth curves. Mean values and SD of three independent experiments are presented. (F), to demonstrated typical Caresponses to ionomycin, DC were pretreated with Fura-2 for 15 minutes prior to the incubation with ionomycin (1 μM). One representative measurement out of four independent experiments is presented