Rapid multiparametric analysis of activation and cytotoxicity in lymphocyte subpopulations (TECH2P.928)

Multiplexing cell health parameters with phenotyping markers provides greater understanding of time and sequence for activation and cellular stress pathways in immune cells. Sample heterogeneity often makes this type of assessment complex to perform and evaluate. The development of the easyCyte™ 12 instruments, which combines 3 lasers and 12 parameters, has greatly facilitated 6-9 color multiparametric analysis of immune cell populations using small amounts of sample without compensation. In this study, we evaluated the impact of activation agents and cytotoxic treatment on PBMCs using a multiparametric assay which identifies lymphocyte subsets in combination with cell health or activation parameters. Our studies demonstrate that overnight treatment with anisomycin resulted in more sensitive impacts on mitochondrial depolarization on all lymphocyte subsets with lower apoptotic and cell death effects being observed. Overnight treatment with celastrol demonstrated almost parallel changes in mitochondrial potential and annexin V based apoptosis effects in lymphocyte subpopulations. The methodology was also utilized to study dose and time course treatments of activation, cellular stress, and cell death caused by PHA or PMA/ionomycin with PBMCs. Which allowed for dissection of sequence of changes in multiple subpopulations. Such studies can greatly enhance our understanding on response of immune cell subpopulations to compounds that cause activation or cytotoxicity.