OMIP‐099: 31‐color spectral flow cytometry panel to investigate the steady‐state phenotype of human T cells

• Published in: Cytometry. Part A Vol. 105; no. 1; pp. 10 - 15
• Main Authors: Zacharias, Zeb R., Houtman, Jon C. D.
• Format: Journal Article
• Language: English
• Published: Hoboken, USA John Wiley & Sons, Inc 01.01.2024; Wiley Subscription Services, Inc
• Subjects: Blood; CC chemokine receptors; CD11a antigen; CD25 antigen; CD27 antigen; CD28 antigen; CD38 antigen; CD4 antigen; CD4 T cells; CD45RA antigen; CD49d antigen; CD56 antigen; CD8 antigen; CD8 T cells; CD95 antigen; Color; Cryopreservation; CXCR3 protein; CXCR5 protein; cytek aurora; Flow Cytometry; Foxp3 protein; gamma‐delta T cells; Humans; Leukocyte Common Antigens - metabolism; Leukocytes (mononuclear); Leukocytes, Mononuclear - metabolism; Lymphocytes; Lymphocytes T; Peripheral blood mononuclear cells; Phenotype; Phenotypes; spectral flow cytometry; T cells; T-Lymphocyte Subsets; T-Lymphocytes - metabolism; gamma-delta T cells; CD4 T cells; T cells; spectral flow cytometry; CD8 T cells; cytek aurora
• ISSN: 1552-4922; 1552-4930; 1552-4930
• DOI: 10.1002/cyto.a.24799

We have developed a 31‐color panel to define the steady‐state phenotype of T cells in human peripheral blood (Table 1). The panel presented here was optimized using cryopreserved peripheral blood mononuclear cells (PBMC). The markers included in this panel were chosen in order to characterize the steady‐state phenotype of T cells and includes markers (CD45RA, CD45RO, CCR7, CD95) to distinguish the main subsets (e.g., naïve, TEM, TCM, TEMRA, TSCM etc.) of CD4, CD8, and γδ T cells. This panel also includes markers for the identification of differentiation status (CD27, CD28), activation/antigen experience status (CD11a, CD49d, CD38, HLA‐DR, CD56, and CD39), co‐inhibitory marker expression (PD‐1, TIM‐3), and CD4 T helper subsets (CXCR3, CXCR5, CCR4, CCR6, Foxp3, CD25, and CD127). This optimized panel provides a broad assessment of the steady‐state phenotype of human T cells.