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Updated: Sep 5, 2025
Single-cell Analysis of Immunophenotype and Cytokine Production in Peripheral Whole Blood via Mass Cytometry
Published on: June 26, 2018
Mass Cytometry and Single-Cell Transcriptome Analyses Reveal the Immune Cell Characteristics of Ulcerative Colitis
Yongxin Luo1, Shiying Liu1, Huibiao Li2
1School of Basic Medical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, China.
Insights
This study identified novel immune cell subsets that differentiate active ulcerative colitis (UCa) from inactive ulcerative colitis (UCin) and healthy controls. These findings may aid in distinguishing between UCa and UCin patient groups.
Area of Science:
- Immunology
- Gastroenterology
- Cell Biology
Background:
- Ulcerative colitis (UC) pathogenesis is intrinsically linked to immune system dysregulation.
- Key immune characteristic differences between active UC (UCa) and inactive UC (UCin) remain incompletely understood.
- Advanced single-cell technologies are crucial for dissecting complex immune profiles in disease states.
Purpose of the Study:
- To elucidate distinct immune cell subset profiles differentiating active UC, inactive UC, and healthy controls.
- To identify novel immune biomarkers for distinguishing between active and inactive ulcerative colitis.
- To leverage high-dimensional cytometry and transcriptomics for comprehensive immune cell characterization.
Main Methods:
- Mass cytometry (CyTOF) was employed to analyze and distinguish immune cell subsets in UCa, UCin, and healthy control (HC) subjects.
- Single-cell RNA sequencing (scRNA-seq) was utilized to validate CyTOF findings and provide deeper molecular insights.
- Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed on differential immune cell subsets.
Main Results:
- Distinct immune cell subsets, including specific effector memory Tregs, B cells, dendritic cells (DCs), and natural killer (NK) cells, were identified.
- UCa and UCin showed enrichment of CCR6+TNF+CD161+ effector memory T cells compared to HC.
- UCa exhibited unique increases in specific Tregs, B cells, macrophages/monocytes, DCs, and cytotoxic NK cells, alongside decreases in plasmablasts and regulatory/tolerant NK cells.
Conclusions:
- Novel immune cell subsets capable of distinguishing between UCa, UCin, and HC individuals were identified.
- The identified immune signatures offer potential for precise differentiation between active and inactive ulcerative colitis patients.
- These findings contribute to a deeper understanding of UC immunopathogenesis and potential therapeutic targets.
Abstract:
Background: The pathogenesis of ulcerative colitis (UC) is closely related to immunity. The immune characteristic differences between active UC (UCa) and inactive UC (UCin) have not been completely explained. Mass cytometry (CyTOF) and single-cell RNA sequencing (scRNA-seq) were used to analyze the immune cells of UCa, UCin and healthy control (HC) subjects to determine the specific immune characteristics. Methods: The immune cell subsets among UCa, UCin, HC were distinguished using CyTOF analysis. scRNA-seq analysis was used to validate the results of CyTOF. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed to understand the roles of differential immune cell subsets. Results: After CyTOF analysis and validation of scRNA-seq analysis, differential immune cell subsets mainly contained TNF+IL-17A++ effector memory (EM) Tregs, CXCR3+CTLA4+ EM Tregs, CXCR3++CCR7+ B cells, HLA-DR+CCR7+ dendritic cells (DCs) and CTLA-4+ natural killer (NK) cells. In comparison to HC, CCR6+TNF+CD161+ EM T cells were highly enriched in UCa and UCin. Besides, UCa was characterized by an increase in CD38+TNF+ EM Tregs, CXCR3+CCR4+ naïve B cells, HLA-DR+CD14+IL21+ macrophages/monocytes, HLA-DR+CCR7+ DCs, AHR+CD14+ cytotoxic NK (cNK) cells and CD8A+IFNG+ cNK cells. Decreases in CD38+CD27+ plasmablasts, CXCR3+CD38+ regulatory NK cells, and CXCR3+CCR7+ tolerant NK cells in UCa were discovered. Conclusions: Novel immune cell subsets which was used to distinguish UCa, UCin and HC were identified. This information might be utilized to distinguish the patients with UCa and UCin.
