OMIP 071: A 31‐Parameter Flow Cytometry Panel for In‐Depth Immunophenotyping of Human T‐Cell Subsets Using Surface Markers

• Published in: Cytometry. Part A Vol. 99; no. 3; pp. 273 - 277
• Main Authors: Wang, Song‐Rong, Zhong, Na, Zhang, Xin‐Mei, Zhao, Zhi‐Bin, Balderas, Robert, Li, Liang, Lian, Zhe‐Xiong
• Format: Journal Article
• Language: English
• Published: Hoboken, USA John Wiley & Sons, Inc 01.03.2021; Wiley Subscription Services, Inc
• Subjects: CC chemokine receptors; CCR6 protein; CCR7 protein; CD103 antigen; CD28 antigen; CD57 antigen; CD69 antigen; CD7 antigen; CD95 antigen; Cell activation; Cell differentiation; Cell surface; Chemokine receptors; Chemokines; CXCR3 protein; CXCR4 protein; CXCR5 protein; Differentiation (biology); Flow cytometry; Helper cells; Histocompatibility antigen HLA; human T‐cell subsets cell surface staining31‐parameter flow cytometry panelPBMCs; Lymph nodes; Lymphocytes; Lymphocytes T; Markers; Parameters; Peripheral blood; Receptors; Surface markers; T cell receptors; Tissue analysis; human T-cell subsets cell surface staining31-parameter flow cytometry panelPBMCs
• DOI: 10.1002/cyto.a.24272
• ISSN: 1552-4922; 1552-4930

Dissecting the functional diversity of T cells is critical in elucidating mechanisms and in developing therapies for various diseases. Here, we designed a 31‐parameter (29‐color) panel to enable the characterization of T‐cell subsets and immunophenotyping of the human peripheral blood and lymph nodes using cell surface staining. In addition to adaptive T‐cell markers, TCR Vα24‐Jα18, TCR γδ, TCR Vɑ7.2, and CD161 were included to identify iNKT, γδ T, and MAIT cells, respectively, which are innate‐like T cells. C‐X‐C chemokine receptors (CXCR3, CXCR4, CXCR5, CXCR6) and C‐C motif chemokine receptors (CCR4, CCR6, CCR7) were included to enable the identification of Th cell subsets (Th1, Th2, Th17), Tfh cell subsets (Tfh1, Tfh2, Tfh17), and Th cells with specific homing capacities. Furthermore, in this panel, we also used markers for assessing cell differentiation (CD45RO, CD7), activation (CD57, CD95, HLA‐DR) and the expression of some cosignaling molecules (PD‐1, NKG2D, CD28). Particularly, CD69 and CD103 were included for the further analysis of tissue resident memory T (Trm) cells. This panel would enable the in‐depth immunophenotyping of human T‐cell subsets, and may be applied in the monitoring, prognosis, and mechanistic studies of various immune‐related diseases.