Proceedings 31st Symposium ESVN-ECVN

In earlier studies an ELISA test measuring IL17 in cerebrospinal fluid (CSF) samples and an ELISpot technique were successfully applied. However, these techniques cannot be performed on a daily routine basis. Therefore, Th17 cells should be measured using flow cytometry. Since Th17 cells are only a minor population of lymphocytes, stimulation is necessary for better visibility. The measurement protocol time takes about 15 hours being the main difficulty for routine analysis. For this reason, results of 10 blood samples divided in two portions were compared: half of the blood sample was processed immediately and the other half was stored overnight for 15‐20 hours. Cells were stimulated with PMA, Ionomycin and Brefeldin A for 6 hours and Th17 cells were intracellularly stained with biotinylated IL17A (Dendritics, France) and measured by flow cytometry.