Abstract Text: Interrogating immune cell composition and function is critical for disease prognoses, monitoring immunotherapies, and identifying novel biomarkers and therapeutic targets. CyTOF® is high-plex flow cytometry that exploits metal-isotope-tagged antibodies. In contrast to fluorescence-based cytometers, CyTOF is not limited by fluorescence overlap, enabling rapid design of 40-plus-marker panels. To expand the clinical and preclinical utility of a 30-marker Maxpar® Direct™ Immune Profiling Assay™ (Maxpar Direct Assay), nine Expansion Panels were developed for deeper phenotyping of cell types and activation states, including markers to characterize ex vivo and activated myeloid cells, T cells, and NK cells.
Peripheral blood mononuclear cells from healthy donors and donors with multiple myeloma were stimulated in vitro, then stained in the 30-antibody Maxpar Direct Assay tube with the NK Cell Expansion Panel (NKp30, NKp46, CD181, PD-1, NKG2A, ICOS, and TIGIT) or the T Cell Expansion Panel 3 (OX40, TIGIT, CD69, PD-1, Tim-3, ICOS, and 4-1BB). Surface staining was followed by intracellular staining with the Basic Activation Expansion Panel antibodies (IL-2, TNFα, IFNγ, perforin, granzyme B) after cell fixation and permeabilization. Anti-CD107a was added during stimulation to measure degranulation. Samples were acquired on a CyTOF XT™ instrument in automated batch acquisition mode. Automated analysis was performed with Maxpar Pathsetter™ software to quantify immune cell activation, cytokine responsiveness and marker expression.
Maxpar reagents and analysis tools thus provide an end-to-end workflow to characterize immune cells in health and disease.
For Research Use Only. Not for use in diagnostic procedures.
