Investigating the potential of full spectral flow cytometry for natural killer cell immunophenotyping

Investigating the potential of full spectral flow cytometry for natural killer cell immunophenotyping

Natural Killer (NK) cells are essential effector cells in mediating protection against viral and cancerous diseases. Full spectrum flow cytometry (FSFC) has recently emerged as a promising new single cell level analysis tool that can be exploited to interrogate the extensive diversity of NK cells and their clinical implication in human diseases. With the increasing complexity of panels that can characterize the deep phenotyping of the immune cell landscape using single cell technologies requires a proper design, a certain layer of optimization, and a well-established standard operating procedure. In this study, we developed a 35 – color panel encompassing major activating and inhibitory NK cell targets with a specific focus on Killer – cell immunoglobulin – like receptors (KIRs). We further cross validated our results by proceeding to a comparative assessment with the single-cell mass cytometry by time-of-flight (CyTOF). Our data revealed an overall high concordance between the platforms, with minor disagreements in the detection and quantification of rare NK cell subpopulations. Our findings demonstrate that FSFC is a highly suitable method to capture the diversity of NK cells and substantiate that the choice of technology is not a primary factor for successful biological assessment of cell profiles but must be considered in a broader framework of clinical studies.