Integrated Proteomics and Single-Cell Mass Cytometry Analysis Dissects the Immune Landscape of Ankylosing Spondylitis
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Updated: Jul 31, 2025
Single-cell Analysis of Immunophenotype and Cytokine Production in Peripheral Whole Blood via Mass Cytometry
Published on: June 26, 2018
Hao Wang1,2, Fengting Luo3, Xianfeng Shao4
1Department of Clinical Laboratory, The Affiliated Cancer Hospital of Zhengzhou University and Henan Cancer Hospital, Zhengzhou 450008, China.
Insights
Integrating mass spectrometry proteomics with mass cytometry reveals immune cell changes in ankylosing spondylitis (AS). This approach identified key protein alterations and cellular interactions, offering insights into AS pathogenesis and potential therapeutic targets.
Area of Science:
- Immunology
- Proteomics
- Single-cell analysis
Background:
- Mass cytometry is a key single-cell technology for immune cell analysis but is limited to a few dozen proteins.
- Understanding immune cell heterogeneity in diseases like ankylosing spondylitis (AS) requires deeper molecular insights.
- Current methods necessitate prior knowledge of specific protein markers for analysis.
Purpose of the Study:
- To integrate mass spectrometry-based proteomics with single-cell mass cytometry for a comprehensive immune system analysis.
- To investigate the immune landscape of ankylosing spondylitis (AS) by identifying proteomic and cellular alterations.
- To uncover potential therapeutic targets and cellular phenotypes for AS treatment.
Main Methods:
- Quantitative proteomics was used to analyze proteome alterations in peripheral blood mononuclear cells (PBMCs) from AS patients.
- Mass cytometry was employed to decipher the immunome, focusing on differentially expressed signaling molecules identified by proteomics.
- Correlation analysis was performed to understand cellular interactions within immune cell subclusters.
Main Results:
- A wide spectrum of dysregulated proteins was identified in AS, including upregulated glycolytic enzymes and downregulated lipid transporters.
- Chemokine signaling molecules involved in inflammation and leukocyte migration were found to be dysregulated.
- Single-cell analysis revealed upregulated chemokine signaling regulators in innate and adaptive immune cell subclusters in AS, alongside interplay between monocyte, CD4+ T cell, and CD8+ T cell subclusters.
Conclusions:
- The integration of mass spectrometry proteomics and single-cell mass cytometry is a powerful strategy for in-depth immune system understanding.
- This combined approach revealed clinically relevant information about immune cell phenotypes and protein alterations in AS.
- The findings provide a foundation for developing novel therapeutic strategies targeting identified pathways and cell populations in AS.
Abstract:
Mass cytometry is a powerful single-cell technology widely adopted to depict immune cell heterogeneity in different contexts. However, this method is only capable of examining several dozens of proteins simultaneously and requires a prior knowledge of the markers to be analyzed. Here we propose that the integration of mass cytometry with shot-gun proteomics may serve as a valuable tool to achieve an in-depth understanding of the immune system. By implementing such a strategy, we investigated the immune landscape of ankylosing spondylitis (AS), a chronic inflammatory arthritis with unclear etiology. The proteome alteration in peripheral blood mononuclear cells (PBMCs) was investigated by quantitative proteomics, and then mass cytometry analysis was conducted to decipher the immunome by considering the signaling molecules identified with differential expression by proteomics. As a result, we identified a wide spectrum of proteins dysregulated in AS, e.g., upregulation of glycolytic enzymes, downregulation of lipid transporters, and dysregulation of chemokine signaling molecules involved in proinflammatory cytokine production and leucocyte migration. Moreover, the single-cell analysis showed the upregulation of chemokine signaling regulators in subclusters of both innate and adaptive immune cells in AS. In addition, correlation analysis unveiled the interplay among Phenograph-identified subclusters of monocytes, CD4+ T cells, and CD8+ T cells. Taken together, our findings demonstrated that the integration of mass spectrometry-based proteomics and single-cell mass cytometry may serve as a useful tool to reveal clinically relevant information regarding useful targets and cellular phenotypes that could be further exploited to develop novel therapeutic strategies.
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