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Updated: Jun 3, 2026
Flow Cytometric Analysis of Lymphocyte Infiltration in Central Nervous System during Experimental Autoimmune Encephalomyelitis
Published on: November 17, 2020
Multiparameter flow cytometry of CSF identifies elevated CD8+ effector memory and TEMRA T-cells in immune-mediated
Danwei Wu1, Samuel Zhang2, Yaseen Ali Jamal3
1Division of Neuroimmunology, Department of Neurology and Neurological Sciences, Stanford University, Stanford, CA, United States.
Insights
Immune-mediated neurologic disorders (IMNDs) can be distinguished from others using cerebrospinal fluid (CSF) CD8+ T-cell effector memory profiles. This T-cell signature may improve diagnosis of CNS autoimmunity.
Area of Science:
- Neuroimmunology
- T-cell immunology
- Cerebrospinal fluid (CSF) analysis
Background:
- Distinguishing immune-mediated neurologic disorders (IMNDs) from non-immune-mediated neurologic disorders (N-IMNDs) using conventional CSF markers is challenging.
- T-cell subsets in the CSF may offer unique insights into CNS autoimmunity.
Purpose of the Study:
- To identify T-cell signatures in CSF associated with IMNDs using multiparametric flow cytometry.
- To evaluate the potential of CSF T-cell profiles as biomarkers for differentiating IMNDs from N-IMNDs.
Main Methods:
- Multiparametric flow cytometric profiling of CSF T-cell developmental subsets.
- Analysis of 37 IMND patients and 10 N-IMND controls.
- Hierarchical clustering and paired blood-CSF analysis in treatment-naïve IMND patients.
Main Results:
- CSF CD8+ T-cells were detected in 86% of IMND patients but not in N-IMND controls (P<0.001).
- Effector memory and terminally differentiated effector memory T-cells (TEMRA) predominated among CSF CD8+ T-cells in IMNDs.
- Hierarchical clustering separated IMND and N-IMND groups based on CSF CD8+ T-cell profiles (P=0.001).
- Paired analysis revealed compartmentalized CNS-restricted CD8+ effector memory T-cell enrichment in IMND patients.
Conclusions:
- CSF CD8+ effector memory T-cell profiles represent a potential biomarker for distinguishing IMNDs from N-IMNDs.
- This T-cell signature may complement existing biomarkers for diagnosing CNS autoimmunity.
- Further validation is warranted to establish the clinical utility of these findings.
Abstract:
Conventional CSF markers often fail to distinguish immune-mediated neurologic disorders (IMNDs) from non-immune-mediated neurologic disorders (N-IMNDs). We performed multiparametric flow cytometric profiling of CSF T-cell developmental subsets in 37 IMND patients and 10 N-IMND controls to identify IMND-associated T-cell signatures. CSF CD8+ T-cells were detectable in 86% (32/37) of IMND patients versus 0% (0/10) of N-IMND controls (Padj < .001). Among CD8+ T-cell-positive IMND cases, effector memory (CD45RA-CCR7-, median, 65.0%; IQR, 45.5%-73.5%) and terminally differentiated effector memory T-cells (TEMRA, CD45RA+CCR7-, median, 35.0%; IQR, 0%-49.5%) predominated. Hierarchical clustering demonstrated significant separation between IMND and N-IMND driven by CD8+ T-cell subset profiles (R2 = 0.165, P = .001), whereas CD4+ T-cell subsets showed no disease-associated clustering. Paired blood-CSF analysis in 6 treatment-naïve IMND patients revealed compartmentalized enrichment of CD8+ effector memory (blood median 19.2%; IQR, 6.4%-26.9%; CSF median 47.3%; IQR, 42.2%-53.7%; P = .031), confirming CNS-restricted CD8+ T-cell activation. These findings identify CSF CD8+ effector memory profiles as a potential biomarker distinguishing IMNDs from N-IMNDs that may complement conventional biomarkers for CNS autoimmunity.
