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Updated: May 7, 2026
Isolation of Cortical Microglia with Preserved Immunophenotype and Functionality From Murine Neonates
Published on: January 30, 2014
Single-cell transcriptome reveals highly complement activated microglia cells in association with pediatric
Siwei Mo1,2, Chenyan Shi2,3, Yi Cai2
1Department of Respiratory Medicine, Children's Hospital of Chongqing Medical University, National Clinical Research Center for Child Health and Disorders, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Children's Hospital of Chongqing Medical University, Chongqing Key Laboratory of Pediatrics, Chongqing, China.
Background:
Tuberculous meningitis (TBM) is a devastating form of tuberculosis (TB) causing high mortality and disability. TBM arises due to immune dysregulation, but the underlying immune mechanisms are unclear.
Methods:
We performed single-cell RNA sequencing on peripheral blood mononuclear cells (PBMCs) and cerebrospinal fluid (CSF) cells isolated from children (n=6) with TBM using 10 xGenomics platform. We used unsupervised clustering of cells and cluster visualization based on the gene expression profiles, and validated the protein and cytokines by ELISA analysis.
Results:
We revealed for the first time 33 monocyte populations across the CSF cells and PBMCs of children with TBM. Within these populations, we saw that CD4_C04 cells with Th17 and Th1 phenotypes and Macro_C01 cells with a microglia phenotype, were enriched in the CSF. Lineage tracking analysis of monocyte populations revealed myeloid cell populations, as well as subsets of CD4 and CD8 T-cell populations with distinct effector functions. Importantly, we discovered that complement-activated microglial Macro_C01 cells are associated with a neuroinflammatory response that leads to persistent meningitis. Consistently, we saw an increase in complement protein (C1Q), inflammatory markers (CRP) and inflammatory factor (TNF-α and IL-6) in CSF cells but not blood. Finally, we inferred that Macro_C01 cells recruit CD4_C04 cells through CXCL16/CXCR6.
Discussion:
We proposed that the microglial Macro_C01 subset activates complement and interacts with the CD4_C04 cell subset to amplify inflammatory signals, which could potentially contribute to augment inflammatory signals, resulting in hyperinflammation and an immune response elicited by Mtb-infected tissues.
Insights
Tuberculous meningitis (TBM) involves immune dysregulation. Microglial Macro_C01 cells activate complement and recruit CD4_C04 cells, amplifying inflammation in TBM.
Area of Science:
- Immunology
- Neuroscience
- Infectious Diseases
Background:
- Tuberculous meningitis (TBM) is a severe form of tuberculosis (TB) with high mortality and disability.
- The precise immune mechanisms driving TBM pathogenesis remain unclear.
Purpose of the Study:
- To elucidate the immune cell populations and interactions involved in TBM pathogenesis.
- To identify key cellular players and molecular pathways contributing to neuroinflammation in TBM.
Main Methods:
- Single-cell RNA sequencing of peripheral blood mononuclear cells (PBMCs) and cerebrospinal fluid (CSF) cells from children with TBM.
- Unsupervised clustering, gene expression profiling, and ELISA validation of proteins and cytokines.
Main Results:
- Identified 33 distinct monocyte populations in PBMCs and CSF.
- Discovered enrichment of CD4_C04 (Th17/Th1) and Macro_C01 (microglia) cells in CSF.
- Macro_C01 cells, activated by complement, were linked to neuroinflammation and recruitment of CD4_C04 cells via CXCL16/CXCR6, with elevated inflammatory markers in CSF.
Conclusions:
- Microglial Macro_C01 cells activate complement and interact with CD4_C04 cells, amplifying inflammatory signals in TBM.
- This interaction may drive hyperinflammation and the immune response to *Mycobacterium tuberculosis* infection in the central nervous system.
