T cells modulate the microglial response to brain ischemia
- Institute for Stroke and Dementia Research, University Hospital, LMU Munich, Germany
- Institute of Diabetes and Regeneration Research, Institute of Computational Biology, Helmholtz Zentrum München, Germany
- Department of Neurology, Focus Program Translational Neuroscience (FTN) and Immunotherapy (FZI), RhineMain Neuroscience Network (rmn(2)), University Medical Center of the Johannes Gutenberg University Mainz, Germany
- Munich Cluster for Systems Neurology (SyNergy), Germany
- Biomedical Center (BMC), Faculty of Medicine, LMU Munich, Germany
- Institute of Clinical Neuroimmunology, University Hospital, LMU Munich, Germany
Figures
Figure 1
Lymphocytes influence microglia morphology after stroke.
(A) Schematic of the experimental design: morphological analysis of microglia and transcriptomic profile of sorted microglia were performed in naïve mice or 5 days after stroke in wild-type (WT) and …
Figure 2 with 3 supplements
Lymphocytes influence microglia transcriptional signature.
(A) CD45+CD11b+ cells were sorted from the ipsilateral hemisphere in naïve mice or 5 days after stroke in wild-type (WT) and Rag1−/− (3 mice per condition), and RNA was isolated for single cell RNA …
Figure 2—figure supplement 1
Transcriptomic analysis of microglia isolated from wild-type (WT) and Rag1−/− mice in naïve and stroke conditions.
(A) Uniform manifold approximation and projection (UMAP) plots showing expression of known CD45+CD11b+ myeloid cell marker genes and Louvain-clusters. (B and C) Manifold and clustering of microglia: …
Figure 2—figure supplement 2
Microglia single cell trajectory inference in wild-type (WT) and Rag1−/− mice in naïve and stroke conditions.
(A and B) Microglia single cell trajectory inference describes the evolution of microglia activation from naïve to stroke condition. (A) Partition-based graph abstraction graph (PAGA, right) shows …
Figure 2—figure supplement 3
Immune cell infiltration in wild-type (WT) and Rag1−/− mice after stroke.
(A) Absolute cell count of CD3+ T cells and CD19+ B cells in the ischemic hemisphere (ipsilateral) 5 days after distal occlusion of the middle cerebral artery (dMCAO) in WT mice as quantified by …
Figure 3 with 1 supplement
TH1 and regulatory T cells (TREG) cells influence microglia gene expression after stroke.
(A) Naïve CD4 cells were polarized in vitro to TH1 or TREG phenotype (Figure 3—figure supplement 1A). One million cells (TH1 or TREG cells) or vehicle (control, CT) were injected into the cisterna …
Figure 3—figure supplement 1
T cell polarization in vitro and infarct volumetry in Rag1−/−.
(A) CD4 naïve T cells were polarized in vitro toward TH1 (top row, CD4+Tbet+) or regulatory T cells (TREG; bottom row, CD4+FoxP3+). (B and C) Infarct volume analysis 5 days after distal occlusion of …
Figure 4 with 1 supplement
Acute post-stroke treatment with engineered T cells overexpressing IL-10 modulates microglial activation and ameliorates functional deficit.
(A and B) Flow cytometry analysis and whole skull-brain coronal sections of 106 eGFP+TH1 cells injected into the cisterna magna (CM) of Rag1−/− mice 24 hr after stroke. Samples were collected 4 hr …
Figure 4—figure supplement 1
Localization of polarized T cell and IL-10 plasmid construct.
(A) Gating strategy of CD4+eGFP+ cells analyzed by flow cytometry in the ipsilateral (red) and contralateral (blue) hemispheres (CD45+CD11b–CD19–CD3+CD4+FITC+) in Rag1−/− (top row), C57BL6/J (no …
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