Single-cell transcriptomic profiling reveals a novel signature of necrotizing granulomatous lesions in the lungs of Mycobacterium tuberculosis-infected C3HeB/FeJ mice

Tuberculosis (TB) pathology involves complex immune responses within granulomatous lesions. Using single-cell RNA sequencing, we characterized the cellular compositions of necrotizing granulomatous lesions that developed in the lungs of Mycobacterium tuberculosis-infected C3HeB/FeJ mice. We identifi...

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Main Authors: Shintaro Seto, Shiho Omori, Hajime Nakamura, Minako Hijikata, Naoto Keicho
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-08-01
Series:Frontiers in Immunology
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Online Access:https://www.frontiersin.org/articles/10.3389/fimmu.2025.1624072/full
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author Shintaro Seto
Shiho Omori
Hajime Nakamura
Hajime Nakamura
Minako Hijikata
Naoto Keicho
Naoto Keicho
author_facet Shintaro Seto
Shiho Omori
Hajime Nakamura
Hajime Nakamura
Minako Hijikata
Naoto Keicho
Naoto Keicho
author_sort Shintaro Seto
collection DOAJ
description Tuberculosis (TB) pathology involves complex immune responses within granulomatous lesions. Using single-cell RNA sequencing, we characterized the cellular compositions of necrotizing granulomatous lesions that developed in the lungs of Mycobacterium tuberculosis-infected C3HeB/FeJ mice. We identified 11 distinct major cell types, including phagocytes such as neutrophils and macrophages, and T cells, natural killer cells, B cells, dendritic cells, and plasmacytoid dendritic cells. Among T cells, particularly, Pdcd1+ γδ T cells were detected in necrotizing granulomatous lesions, suggesting their potential role in the pathogenicity of M. tuberculosis. Within the macrophage populations, we identified a cluster with significantly higher Plin2 expression compared to other clusters, whose transcriptomic profile was consistent with that of foamy macrophages. A subset of the Plin2-expressing macrophages was identified as a major source of Ifnb1 and Cxcl1, suggesting their involvement in type I interferon signaling and neutrophil recruitment. Furthermore, we identified Flrt2, Hyal1, and Mmp13 as novel molecular markers of Plin2-expressing macrophages, which were localized to the peripheral rim regions of necrotizing granulomas. In conclusion, our results provide the immune landscape of necrotizing granulomas and reveal novel functional states of macrophages contributing to TB pathogenesis.
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spelling doaj-art-d67ff433f72b4974a36b7ed07e7d464d2025-08-20T03:44:35ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-08-011610.3389/fimmu.2025.16240721624072Single-cell transcriptomic profiling reveals a novel signature of necrotizing granulomatous lesions in the lungs of Mycobacterium tuberculosis-infected C3HeB/FeJ miceShintaro Seto0Shiho Omori1Hajime Nakamura2Hajime Nakamura3Minako Hijikata4Naoto Keicho5Naoto Keicho6Department of Pathophysiology and Host Defense, The Research Institute of Tuberculosis, Japan Anti-Tuberculosis Association, Tokyo, JapanDepartment of Pathophysiology and Host Defense, The Research Institute of Tuberculosis, Japan Anti-Tuberculosis Association, Tokyo, JapanDepartment of Pathophysiology and Host Defense, The Research Institute of Tuberculosis, Japan Anti-Tuberculosis Association, Tokyo, JapanDepartment of Basic Mycobacteriosis, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, JapanDepartment of Pathophysiology and Host Defense, The Research Institute of Tuberculosis, Japan Anti-Tuberculosis Association, Tokyo, JapanDepartment of Basic Mycobacteriosis, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, JapanThe Research Institute of Tuberculosis, Japan Anti-Tuberculosis Association, Tokyo, JapanTuberculosis (TB) pathology involves complex immune responses within granulomatous lesions. Using single-cell RNA sequencing, we characterized the cellular compositions of necrotizing granulomatous lesions that developed in the lungs of Mycobacterium tuberculosis-infected C3HeB/FeJ mice. We identified 11 distinct major cell types, including phagocytes such as neutrophils and macrophages, and T cells, natural killer cells, B cells, dendritic cells, and plasmacytoid dendritic cells. Among T cells, particularly, Pdcd1+ γδ T cells were detected in necrotizing granulomatous lesions, suggesting their potential role in the pathogenicity of M. tuberculosis. Within the macrophage populations, we identified a cluster with significantly higher Plin2 expression compared to other clusters, whose transcriptomic profile was consistent with that of foamy macrophages. A subset of the Plin2-expressing macrophages was identified as a major source of Ifnb1 and Cxcl1, suggesting their involvement in type I interferon signaling and neutrophil recruitment. Furthermore, we identified Flrt2, Hyal1, and Mmp13 as novel molecular markers of Plin2-expressing macrophages, which were localized to the peripheral rim regions of necrotizing granulomas. In conclusion, our results provide the immune landscape of necrotizing granulomas and reveal novel functional states of macrophages contributing to TB pathogenesis.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1624072/fulltuberculosisMycobacterium tuberculosisC3HeB/FeJ TB modelnecrotizing granulomafoamy macrophagescRNA-seq
spellingShingle Shintaro Seto
Shiho Omori
Hajime Nakamura
Hajime Nakamura
Minako Hijikata
Naoto Keicho
Naoto Keicho
Single-cell transcriptomic profiling reveals a novel signature of necrotizing granulomatous lesions in the lungs of Mycobacterium tuberculosis-infected C3HeB/FeJ mice
Frontiers in Immunology
tuberculosis
Mycobacterium tuberculosis
C3HeB/FeJ TB model
necrotizing granuloma
foamy macrophage
scRNA-seq
title Single-cell transcriptomic profiling reveals a novel signature of necrotizing granulomatous lesions in the lungs of Mycobacterium tuberculosis-infected C3HeB/FeJ mice
title_full Single-cell transcriptomic profiling reveals a novel signature of necrotizing granulomatous lesions in the lungs of Mycobacterium tuberculosis-infected C3HeB/FeJ mice
title_fullStr Single-cell transcriptomic profiling reveals a novel signature of necrotizing granulomatous lesions in the lungs of Mycobacterium tuberculosis-infected C3HeB/FeJ mice
title_full_unstemmed Single-cell transcriptomic profiling reveals a novel signature of necrotizing granulomatous lesions in the lungs of Mycobacterium tuberculosis-infected C3HeB/FeJ mice
title_short Single-cell transcriptomic profiling reveals a novel signature of necrotizing granulomatous lesions in the lungs of Mycobacterium tuberculosis-infected C3HeB/FeJ mice
title_sort single cell transcriptomic profiling reveals a novel signature of necrotizing granulomatous lesions in the lungs of mycobacterium tuberculosis infected c3heb fej mice
topic tuberculosis
Mycobacterium tuberculosis
C3HeB/FeJ TB model
necrotizing granuloma
foamy macrophage
scRNA-seq
url https://www.frontiersin.org/articles/10.3389/fimmu.2025.1624072/full
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