Application of imaging mass cytometry for spatially profiling the microenvironment of salivary glands in primary Sjögren’s syndrome

Abstract Primary Sjogren’s syndrome (pSS) is a slowly progressive, systemic autoimmune disorder characterized by gradual lymphocytic infiltration of exocrine glands. However, the spatially profiling the immune microenvironment in pSS is largely unclear, limiting the understanding of the complex inte...

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Main Authors: Guolin Wu, Fangping Wu, Lipei Wang, Lixiong Ying, Wenwen Lu, Kang Qian, Tianxiao Fu, Danbin Wu, Fenglin Hu, YiHang Shi, Li Xu
Format: Article
Language:English
Published: Nature Publishing Group 2025-05-01
Series:Cell Death and Disease
Online Access:https://doi.org/10.1038/s41419-025-07717-7
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author Guolin Wu
Fangping Wu
Lipei Wang
Lixiong Ying
Wenwen Lu
Kang Qian
Tianxiao Fu
Danbin Wu
Fenglin Hu
YiHang Shi
Li Xu
author_facet Guolin Wu
Fangping Wu
Lipei Wang
Lixiong Ying
Wenwen Lu
Kang Qian
Tianxiao Fu
Danbin Wu
Fenglin Hu
YiHang Shi
Li Xu
author_sort Guolin Wu
collection DOAJ
description Abstract Primary Sjogren’s syndrome (pSS) is a slowly progressive, systemic autoimmune disorder characterized by gradual lymphocytic infiltration of exocrine glands. However, the spatially profiling the immune microenvironment in pSS is largely unclear, limiting the understanding of the complex interplay among cells within the microenvironment. Based on imaging mass cytometry (IMC) analysis of clinical pSS samples, we first revealed that labial salivary gland (LSG) comprised of epithelial, immune cells and stromal cells, and epithelial was the main cell type in LSG. Eight immune cells populations were identified, including CD8+ T, CD4+ T, Treg, B, NK cells, neutrophils, resident macrophages and a mixed immune cell cluster. We found that CD8+ T cells, but not CD4+ T cells, were the most prominent T cells in immune infiltrates of pSS LSG. With the increase of pSS disease activity and severity, the infiltration abundance of CD8+ T cells gradually increased and was accompanied by the activation of inflammatory response. sc-RNA-seq analysis based on the GSE272409 dataset confirmed that CD8+ T cells were the main immune cells, and dominated the most intercellular ligand-receptor interactions. CD8+ T cells were further clustered into five cell subsets, of which CD160+CD8+ T cells subset appeared to present only in pSS patients. Further experiments demonstrated that CD160 expression on CD8+ T cells was associated with an enhanced expression of proinflammatory and cytotoxic cytokines IFN-γ, GZMB and TNF-α, and the injury of salivary gland epithelial cells. Besides, proportion of GZMK+CD8+ T cells subset was increased in pSS patients. Trajectory analysis confirmed an enhanced frequency of CD8+ T cell differentiation and activation during the progression of pSS. This study provided single cell profile with spatial information for analyzing the LSG immune microenvironment in pSS, which could not be achieved by conventional immunofluorescence and immunohistochemistry assays.
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spelling doaj-art-e76d7cf89e664ff099f4cfa22911eb6f2025-08-20T03:10:32ZengNature Publishing GroupCell Death and Disease2041-48892025-05-0116111610.1038/s41419-025-07717-7Application of imaging mass cytometry for spatially profiling the microenvironment of salivary glands in primary Sjögren’s syndromeGuolin Wu0Fangping Wu1Lipei Wang2Lixiong Ying3Wenwen Lu4Kang Qian5Tianxiao Fu6Danbin Wu7Fenglin Hu8YiHang Shi9Li Xu10Department of Traditional Chinese Medicine, The First Affiliated Hospital, Zhejiang University School of MedicineSchool of Pharmaceutical Sciences, Zhejiang Chinese Medical UniversityHangzhou Normal University of Basic Medical SciencesThe First Affiliated Hospital, Zhejiang University School of MedicineDepartment of Traditional Chinese Medicine, The First Affiliated Hospital, Zhejiang University School of MedicineDepartment of Traditional Chinese Medicine, The First Affiliated Hospital, Zhejiang University School of MedicineDepartment of Traditional Chinese Medicine, The First Affiliated Hospital, Zhejiang University School of MedicineDepartment of Traditional Chinese Medicine, The First Affiliated Hospital, Zhejiang University School of MedicineDepartment of Traditional Chinese Medicine, The First Affiliated Hospital, Zhejiang University School of MedicineDepartment of Traditional Chinese Medicine, The First Affiliated Hospital, Zhejiang University School of MedicineDepartment of Traditional Chinese Medicine, The First Affiliated Hospital, Zhejiang University School of MedicineAbstract Primary Sjogren’s syndrome (pSS) is a slowly progressive, systemic autoimmune disorder characterized by gradual lymphocytic infiltration of exocrine glands. However, the spatially profiling the immune microenvironment in pSS is largely unclear, limiting the understanding of the complex interplay among cells within the microenvironment. Based on imaging mass cytometry (IMC) analysis of clinical pSS samples, we first revealed that labial salivary gland (LSG) comprised of epithelial, immune cells and stromal cells, and epithelial was the main cell type in LSG. Eight immune cells populations were identified, including CD8+ T, CD4+ T, Treg, B, NK cells, neutrophils, resident macrophages and a mixed immune cell cluster. We found that CD8+ T cells, but not CD4+ T cells, were the most prominent T cells in immune infiltrates of pSS LSG. With the increase of pSS disease activity and severity, the infiltration abundance of CD8+ T cells gradually increased and was accompanied by the activation of inflammatory response. sc-RNA-seq analysis based on the GSE272409 dataset confirmed that CD8+ T cells were the main immune cells, and dominated the most intercellular ligand-receptor interactions. CD8+ T cells were further clustered into five cell subsets, of which CD160+CD8+ T cells subset appeared to present only in pSS patients. Further experiments demonstrated that CD160 expression on CD8+ T cells was associated with an enhanced expression of proinflammatory and cytotoxic cytokines IFN-γ, GZMB and TNF-α, and the injury of salivary gland epithelial cells. Besides, proportion of GZMK+CD8+ T cells subset was increased in pSS patients. Trajectory analysis confirmed an enhanced frequency of CD8+ T cell differentiation and activation during the progression of pSS. This study provided single cell profile with spatial information for analyzing the LSG immune microenvironment in pSS, which could not be achieved by conventional immunofluorescence and immunohistochemistry assays.https://doi.org/10.1038/s41419-025-07717-7
spellingShingle Guolin Wu
Fangping Wu
Lipei Wang
Lixiong Ying
Wenwen Lu
Kang Qian
Tianxiao Fu
Danbin Wu
Fenglin Hu
YiHang Shi
Li Xu
Application of imaging mass cytometry for spatially profiling the microenvironment of salivary glands in primary Sjögren’s syndrome
Cell Death and Disease
title Application of imaging mass cytometry for spatially profiling the microenvironment of salivary glands in primary Sjögren’s syndrome
title_full Application of imaging mass cytometry for spatially profiling the microenvironment of salivary glands in primary Sjögren’s syndrome
title_fullStr Application of imaging mass cytometry for spatially profiling the microenvironment of salivary glands in primary Sjögren’s syndrome
title_full_unstemmed Application of imaging mass cytometry for spatially profiling the microenvironment of salivary glands in primary Sjögren’s syndrome
title_short Application of imaging mass cytometry for spatially profiling the microenvironment of salivary glands in primary Sjögren’s syndrome
title_sort application of imaging mass cytometry for spatially profiling the microenvironment of salivary glands in primary sjogren s syndrome
url https://doi.org/10.1038/s41419-025-07717-7
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