30-color full spectrum flow cytometry panel for deep immunophenotyping of T cell subsets in murine tumor tissue

Ying Liu; Xiye Xu; Di Liu; Xiayan Wu; Yajing Gao; Huan Wang; Fang Yan; Wenting Yang; Dianyuan Zhao; Fuchu He; Li Tang

doi:10.1016/j.jim.2023.113459

30-color full spectrum flow cytometry panel for deep immunophenotyping of T cell subsets in murine tumor tissue

J Immunol Methods. 2023 May:516:113459. doi: 10.1016/j.jim.2023.113459. Epub 2023 Mar 16.

Authors

Ying Liu¹, Xiye Xu¹, Di Liu¹, Xiayan Wu¹, Yajing Gao², Huan Wang¹, Fang Yan¹, Wenting Yang¹, Dianyuan Zhao¹, Fuchu He³, Li Tang⁴

Affiliations

¹ State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences·Beijing, Beijing Institute of Lifeomics, Beijing, China.
² Cytek (Shanghai) BioSciences Co.Ltd 603, No.560 ShengXia Road, Z.J. Hi-Tech Park, Shanghai 201203, China.
³ State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences·Beijing, Beijing Institute of Lifeomics, Beijing, China; Research Unit of Proteomics Driven Cancer Precision Medicine, Chinese Academy of Medical Sciences, Beijing 102206, China.
⁴ State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences·Beijing, Beijing Institute of Lifeomics, Beijing, China. Electronic address: tangli@ncpsb.org.cn.

PMID: 36931458
DOI: 10.1016/j.jim.2023.113459

Abstract

This 30-color full spectrum flow cytometry panel was developed and optimized for in-depth analysis T cells immunophenotype in tumor microenvironment and peripheral lymphoid organs. The panel presented here first identify the main cell subsets including myeloid cells, B cells, NKT cells, γδ T cells, CD4⁺ T cells and CD8⁺ T cells. For CD4⁺ T cells or CD8⁺ T cells, the panel includes markers for further characterization by including a selection of activation status(CD44, CD62L, CD69, Ki67, CD127, KLRG1 and CXCR3), costimulatory/co-inhibitory molecules (ICOS, OX-40, PD-1, LAG3, TIM-3, CTLA-4 and TIGIT), pro-inflammatory/anti-inflammatory cytokines (IFN-γ, TNF-α and IL-10) and cytotoxic molecules (Perforin, Granzymes B and CD107a). The panel has been tested on the tumor infiltrating T cells and corresponding spleen T cells in B16-F10 murine melanoma models.

Keywords: Full spectrum; High-dimensional flow cytometry; T cell immunophenotyping; Tumor microenvironment.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
CD8-Positive T-Lymphocytes*
Cytokines
Flow Cytometry
Immunophenotyping
Melanoma, Experimental* / diagnosis
Mice
T-Lymphocyte Subsets
Tumor Microenvironment

Substances

Cytokines