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Updated: Dec 5, 2025
Isolation and Flow Cytometric Assessment of Neuroimmune Interactions in a Mini-Stroke Murine Model
Published on: June 20, 2025
CD200-CD200R1 signaling pathway regulates neuroinflammation after stroke
Shou-Cai Zhao1, Xu Heng2, Wang Ya-Ping1
1Department of Neurology, Wannan Medical College, Yijishan Hospital, Wuhu, China.
Objective:
To study how the CD200-CD200R1 signaling pathway modulates poststroke inflammation and advances our knowledge of immune responses to ischemia insults in stroke.
Methods:
Focal middle cerebral artery occlusion (MCAO) was induced in mice for 90 min, and mice were sacrificed at 1, 3, and 7 days of reperfusion. CD200, CD200R1, iNOS, and Arg-1 expression in ischemic brains was assessed by Western blotting (WB), and immunohistochemical (IHC) staining was performed to examine the expression of CD200 on neurons and CD200R1 on infiltrating lymphocytes. The severity of neurobehavioral deficits was evaluated by neurological deficit scores (NDS) and infarction volume estimated by TTC staining. To study the relationship between CD200/CD200R1 expression and the diversity of the neuroinflammatory response in stroke, CD200Fc (CD200R1 agonist) was subcutaneously injected at onset, at 1 day and 2 days after MCAO operation, and the brains were collected for detection at 3 days after MCAO/R (reperfusion).
Results:
CD200 expression on neurons increased at 1 day and then decreased at 3 days after MCAO/R, and the expression of CD200R1 on lymphocytes showed an opposite temporal pattern as tested by IHC. The WB results showed that CD200/CD200R1 variance exhibited a similar pattern of IHC results, and the level of iNOS peaked at 1 day and then decreased gradually, but Arg-1 increased with time after MCAO/R in ischemic brains. After CD200Fc injection, CD200R1 expression significantly increased, and CD200Fc promoted Arg-1 but inhibited iNOS expression. The infarct volume and NDS of the group treated with CD200Fc were significantly smaller than those of the IgG2a-treated group.
Conclusions:
The CD200-CD200R1 signaling pathway regulates neuroinflammation after stroke. Stimulation of CD200R1 by CD200Fc promotes the anti-inflammatory response and alleviates ischemic injury.
Insights
The CD200-CD200R1 pathway regulates brain inflammation after stroke. Activating CD200R1 with CD200Fc reduces inflammation and lessens ischemic injury, offering a potential therapeutic target for stroke recovery.
Area of Science:
- Neuroscience
- Immunology
- Stroke Research
Background:
- Stroke triggers neuroinflammation, a complex immune response impacting recovery.
- The CD200-CD200R1 signaling pathway plays a role in immune regulation.
Purpose of the Study:
- To investigate how the CD200-CD200R1 pathway modulates post-stroke inflammation.
- To advance understanding of immune responses to ischemic insults in stroke.
Main Methods:
- Middle cerebral artery occlusion (MCAO) model in mice to simulate ischemic stroke.
- Western blotting and immunohistochemistry to assess protein expression (CD200, CD200R1, iNOS, Arg-1).
- Neurobehavioral deficit scores and infarct volume measurements to evaluate stroke severity; CD200Fc administration to study pathway activation.
Main Results:
- CD200 expression on neurons and CD200R1 on lymphocytes showed dynamic temporal changes post-MCAO.
- CD200Fc treatment increased CD200R1 expression, promoted anti-inflammatory Arg-1, and inhibited pro-inflammatory iNOS.
- CD200Fc administration significantly reduced infarct volume and neurological deficits.
Conclusions:
- The CD200-CD200R1 signaling pathway is a key regulator of neuroinflammation following ischemic stroke.
- Stimulating CD200R1 with CD200Fc demonstrates therapeutic potential by promoting anti-inflammatory responses and mitigating ischemic brain injury.
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