<scp>OMIP</scp>‐079: Cell cycle of <scp>CD4</scp><sup>+</sup> and <scp>CD8</scp><sup>+</sup> naïve/memory T cell subsets, and of Treg cells from mouse spleen
Ambra Natalini, Sonia Simonetti, Gabriele Favaretto, Giovanna Peruzzi, Fabrizio Antonangeli, Angela Santoni, Miguel Muñoz‐Ruiz, Adrian Hayday, Francesca Di Rosa
Abstract
Abstract 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: G 0 (Ki67 − , with 2n DNA), G 1 (Ki67 + , with 2n DNA), and S‐G 2 /M (Ki67 + , with 2n < DNA ≤ 4n). This panel was established for the analysis of mouse (C57BL/6J) spleen.