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Updated: Jun 23, 2026
Analysis of Microglia and Monocyte-derived Macrophages from the Central Nervous System by Flow Cytometry
Published on: June 22, 2017
Macrophages of multiple sclerosis patients display deficient SHP-1 expression and enhanced inflammatory phenotype
George P Christophi1, Michael Panos, Chad A Hudson
1Department of Neurology, SUNY Upstate Medical University, Syracuse, NY 13210, USA.
Abstract:
Recent studies in mice have demonstrated that the protein tyrosine phosphatase SHP-1 is a crucial negative regulator of proinflammatory cytokine signaling, TLR signaling, and inflammatory gene expression. Furthermore, mice genetically lacking SHP-1 (me/me) display a profound susceptibility to inflammatory CNS demyelination relative to wild-type mice. In particular, SHP-1 deficiency may act predominantly in inflammatory macrophages to increase CNS demyelination as SHP-1-deficient macrophages display coexpression of inflammatory effector molecules and increased demyelinating activity in me/me mice. Recently, we reported that PBMCs of multiple sclerosis (MS) patients have a deficiency in SHP-1 expression relative to normal control subjects indicating that SHP-1 deficiency may play a similar role in MS as to that seen in mice. Therefore, it became essential to examine the specific expression and function of SHP-1 in macrophages from MS patients. Herein, we document that macrophages of MS patients have deficient SHP-1 protein and mRNA expression relative to those of normal control subjects. To examine functional consequences of the lower SHP-1, the activation of STAT6, STAT1, and NF-kappaB was quantified and macrophages of MS patients showed increased activation of these transcription factors. In accordance with this observation, several STAT6-, STAT1-, and NF-kappaB-responsive genes that mediate inflammatory demyelination were increased in macrophages of MS patients following cytokine and TLR agonist stimulation. Supporting a direct role of SHP-1 deficiency in altered macrophage function, experimental depletion of SHP-1 in normal subject macrophages resulted in an increased STAT/NF-kappaB activation and increased inflammatory gene expression to levels seen in macrophages of MS patients. In conclusion, macrophages of MS patients display a deficiency of SHP-1 expression, heightened activation of STAT6, STAT1, and NF-kappaB and a corresponding inflammatory profile that may be important in controlling macrophage-mediated demyelination in MS.
Insights
Macrophages from multiple sclerosis (MS) patients show reduced SHP-1 protein and mRNA expression. This deficiency leads to increased inflammatory responses and demyelination, suggesting a key role for SHP-1 in MS pathogenesis.
Area of Science:
- Neuroimmunology
- Molecular and Cellular Biology
Background:
- Protein tyrosine phosphatase SHP-1 negatively regulates inflammatory signaling.
- SHP-1 deficiency in mice (me/me) increases susceptibility to central nervous system (CNS) demyelination, particularly in macrophages.
- Peripheral blood mononuclear cells (PBMCs) from multiple sclerosis (MS) patients exhibit reduced SHP-1 expression compared to controls.
Purpose of the Study:
- To investigate the expression and function of SHP-1 in macrophages from MS patients.
- To determine if SHP-1 deficiency in MS patient macrophages contributes to inflammatory demyelination.
Main Methods:
- Quantification of SHP-1 protein and mRNA in macrophages from MS patients and healthy controls.
- Assessment of STAT6, STAT1, and NF-kappaB activation in macrophages.
- Analysis of STAT6-, STAT1-, and NF-kappaB-responsive gene expression following stimulation.
- Experimental depletion of SHP-1 in normal macrophages to mimic MS patient conditions.
Main Results:
- Macrophages from MS patients exhibit significantly lower SHP-1 protein and mRNA levels compared to controls.
- MS patient macrophages show heightened activation of transcription factors STAT6, STAT1, and NF-kappaB.
- Expression of inflammatory genes, mediated by STAT6, STAT1, and NF-kappaB, is increased in MS patient macrophages.
- Experimental SHP-1 depletion in normal macrophages recapitulates the heightened activation and inflammatory gene expression seen in MS patient macrophages.
Conclusions:
- Macrophages from MS patients are characterized by SHP-1 deficiency.
- This deficiency results in increased STAT6, STAT1, and NF-kappaB activation, promoting an inflammatory profile.
- Altered macrophage function due to SHP-1 deficiency may be a critical factor in driving macrophage-mediated demyelination in MS.
