OMIP-079: Cell cycle of CD4⁺ and CD8⁺ naïve/memory T cell subsets, and of Treg cells from mouse spleen

A multicolor flow cytometry panel was designed and optimized to define the following nine mouse T cell subsets: Treg (CD3⁺ CD4⁺ CD8^- FoxP3⁺ ), CD4⁺ T naïve (CD3⁺ CD4⁺ CD8^- FoxP3^- CD44^int/low CD62L⁺ ), CD4⁺ T central memory (CD3⁺ CD4⁺ CD8^- FoxP3^- CD44^high CD62L⁺ ), CD4⁺ T effector memory (CD3⁺ CD4⁺ CD8^- FoxP3^- CD44^high CD62L^- ), CD4⁺ T EMRA (CD3⁺ CD4⁺ CD8^- FoxP3^- CD44^int/low CD62L^- ), CD8⁺ T naïve (CD3⁺ CD8⁺ CD4^- CD44^int/low CD62L⁺ ), CD8⁺ T central memory (CD3⁺ CD8⁺ CD4^- CD44^high CD62L⁺ ), CD8⁺ T effector memory (CD3⁺ CD8⁺ CD4^- CD44^high CD62L^- ), and CD8⁺ T EMRA (CD3⁺ CD8⁺ CD4^- CD44^int/low CD62L^- ). In each T cell subset, a dual staining for Ki-67 expression and DNA content was employed to distinguish the following cell cycle phases: G₀ (Ki67^- , with 2n DNA), G₁ (Ki67⁺ , with 2n DNA), and S-G₂ /M (Ki67⁺ , with 2n < DNA ≤ 4n). This panel was established for the analysis of mouse (C57BL/6J) spleen.