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Midbrain microglia mediate a specific immunosuppressive response under inflammatory conditions
Miguel Angel Abellanas1,2, Marta Zamarbide2, Leyre Basurco1,2
1Departamento de Bioquímica y Genética, Universidad de Navarra, Facultad de Ciencias, Pamplona, Spain.
Background:
Inflammation is a critical process for the progression of neuronal death in neurodegenerative disorders. Microglia play a central role in neuroinflammation and may affect neuron vulnerability. Next generation sequencing has shown the molecular heterogeneity of microglial cells; however, the variability in their response to pathological inputs remains unknown.
Methods:
To determine the effect of an inflammatory stimulus on microglial cells, lipopolysaccharide (LPS) was administered peripherally to mice and the inflammatory status of the cortex, hippocampus, midbrain, and striatum was assessed. Microglial activation and interaction with the immune system were analyzed in single cell suspensions obtained from the different brain regions by fluorescence-activated cell sorting, next generation RNA sequencing, real-time PCR, and immunohistochemical techniques. Antigen-presenting properties of microglia were evaluated by the ability of isolated cells to induce a clonal expansion of CD4+ T cells purified from OT-II transgenic mice.
Results:
Under steady-state conditions, the midbrain presented a high immune-alert state characterized by the presence of two unique microglial subpopulations, one expressing the major histocompatibility complex class II (MHC-II) and acting as antigen-presenting cells and another expressing the toll-like receptor 4 (TLR4), and by the presence of a higher proportion of infiltrating CD4+ T cells. This state was not detected in the cortex, hippocampus, or striatum. Systemic LPS administration induced a general increase in classic pro-inflammatory cytokines, in co-inhibitory programmed death ligand 1 (PD-L1), and in cytotoxic T lymphocyte antigen 4 (CTLA-4) receptors, as well as a decrease in infiltrating effector T cells in all brain regions. Interestingly, a specific immune-suppressive response was observed in the midbrain which was characterized by the downregulation of MHC-II microglial expression, the upregulation of the anti-inflammatory cytokines IL10 and TGFβ, and the increase in infiltrating regulatory T cells.
Conclusions:
These data show that the midbrain presents a high immune-alert state under steady-state conditions that elicits a specific immune-suppressive response when exposed to an inflammatory stimulus. This specific inflammatory tone and response may have an impact in neuronal viability.
Insights
The midbrain exhibits a unique immune-alert state with specific microglial subpopulations. Upon inflammation, it mounts an immune-suppressive response, potentially impacting neuronal survival in neurodegenerative diseases.
Area of Science:
- Neuroscience
- Immunology
Background:
- Neuroinflammation, driven by microglia, is key in neurodegenerative diseases.
- Microglial heterogeneity suggests varied responses to pathology, but this remains unclear.
Purpose of the Study:
- To investigate microglial responses to peripheral inflammatory stimuli.
- To characterize regional differences in microglial activation and immune interactions within the brain.
Main Methods:
- Peripheral lipopolysaccharide (LPS) administration in mice.
- Single-cell analysis (FACS, RNA-seq, qPCR, IHC) of microglia from cortex, hippocampus, midbrain, and striatum.
- Assessment of microglial antigen-presenting capacity using CD4+ T cells.
Main Results:
- Steady-state midbrain microglia show an immune-alert phenotype with MHC-II and TLR4 expression and higher CD4+ T cell infiltration.
- Systemic LPS induced general pro-inflammatory markers and decreased effector T cells across brain regions.
- The midbrain uniquely displayed an immune-suppressive response with downregulated MHC-II, upregulated IL10/TGFβ, and increased regulatory T cells.
Conclusions:
- The midbrain possesses a distinct immune-alert state under normal conditions.
- This state triggers a specific immune-suppressive response to inflammatory challenges.
- This unique neuroinflammatory profile may influence neuronal viability in disease contexts.
