Characterization of murine lymphocyte activation and exhaustion markers by a 14-color flow cytometry panel

Previously designed flow cytometry panels have provided a framework to analyze T-cell activation; however, few provide an extensive view of lymphocyte populations, and none are optimized for murine models. This article describes a panel designed specifically to assess the expression of activation and exhaustion markers in expanding lymphocyte populations in tumor-bearing mice across two distinct genetic backgrounds: BALB/c and C57BL/6. This comprehensive panel enables the assessment of multiple functional states and immune checkpoint markers across cytotoxic CD8⁺ T cells, helper and regulatory CD4⁺ T cells and natural killer cells in murine whole blood, lymph nodes and tumor.