Analysis by flow cytometry of calcium influx kinetics in peripheral lymphocytes of patients with rheumatoid arthritis

To characterize Ca2+ influx kinetics upon activation in major T cell subsets (CD4, Th1, Th2, CD8) isolated from patients with recently diagnosed and established RA and their sensitivity to the specific inhibition of Kv1.3 and IKCa1 lymphocyte K+ channels.
Conclusion:
Th2 cells of patients with established RA react slower to activating stimuli, whereas CD8 cells show a faster reaction than in recently diagnosed RA patients. While initially Th1 cells are less sensitive to the inhibition of Kv1.3 and IKCa1 channels in RA, their sensitivity increases along with the duration of the disease. With the algorithm of function fitting instead of smoothing, more statistically significant differences of potassium channel inhibition between the two RA groups could be demonstrated. The function fitting algorithm applied by FacsKin is suitable to provide a common basis for evaluating and comparing flow cytometry kinetic data. Unstained and single stained PBMC controls were used for pre-experimental setup and for compensation. The flow cytometer was set up before experiments using seven-color beads (BD FACS, BD Biosciences, San Jose, CA, USA). Voltages were adjusted such that fluorescence intensity was identical for each antibody over the experiments.