MBTP 11: Immunophenotyping of T cells from mouse spleen using flow cytometry

Gating strategy showing the analysis of T cell subsets from mouse spleen. Splenocytes from C57BL/6 mice were stained using the previously described T cell panel to identify major T cell subsets. CD4⁺ and CD8⁺ T cells were identified from CD3-expressing cell subsets (A, B). Different subsets of naive, central memory (CM), effector memory (EM), CD127-, and KLRG1-expressing T cells were analyzed within CD4⁺ (C, D, E) and CD8⁺ T cells (F, G, H) respectively. The differential expression of the adhesion molecules CD62L and CD44 was used to identify naive T cells from CM and EM T cells. Naive T cells expressed high levels of CD62L and showed absent or low expression of CD44 (C). Both CM and EM populations expressed CD44. CM T cells also expressed CD62L, whereas expression of CD62L was absent in EM T cells. Analysis of CD127 and KLRG1 was included in this panel to allow the study of T cell responses during infection. During the course of an infection the levels of KLRG1 increase, especially on CD8⁺ T cells.