Immune-profiling of T helper 1 (Th1), Th2 and Th17 signatures in murine splenocytes by targeting intracellular cytokines

Functional cytokines shape both innate and adaptive immune responses in the host after infection or immunization. Deep immunophenotyping of the key functional cytokine signatures associated with T cells in murine lymphoid tissue, especially in the spleen, is challenging. Using spectral flow cytometry, we developed a 17-parameter panel to profile major immune cell subsets along with T cells, memory phenotypes and functional cytokines in murine splenocytes in steady state as well as in stimulated conditions. This panel dissects the memory T cell compartment via CD62L and CD44 expression after mitogen stimulation. To profile T helper (Th) cells distribution after mitogen stimulation, established Th1 markers IFNγ, TNF and IL-2; Th2 markers IL-4/5 and the Th17 marker, IL-17, are included. This optimized multicolor spectral flow panel allows a detailed immune profiling of functional cytokines in the murine T cell compartment and might be useful for exploratory analysis of how these functional cytokines shape host immunity after infection or vaccination. Our panel could be easily modified, if researchers wish to tailor the panel to their specific needs.