Related Experiment Video
Updated: Mar 24, 2026
Author Spotlight: Creating a Versatile Experimental Autoimmune Encephalomyelitis Model Relevant for Both Male and Female Mice
Published on: October 13, 2023
A leading role for NADPH oxidase in an in-vitro study of experimental autoimmune encephalomyelitis
Ji-Eun Seo1, Mahbub Hasan1, Khandoker Asiqur Rahaman1
1Toxicology Laboratory, Doping Control Center, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea; Department of Biological Chemistry, Korea University of Science and Technology, Daejeon 34113, Republic of Korea.
Abstract:
Myelin oligodendrocyte glycoprotein peptide fragment 35-55 (MOG35-55) is a major autoantigen inducing experimental autoimmune encephalomyelitis, an animal model of multiple sclerosis that is characterized by blood-brain barrier (BBB) disruption. Various experimental approaches have employed MOG35-55 in vivo; however, in vitro BBB models using MOG35-55 are rarely reported. We investigated MOG35-55 exposure effects with complete Freund's adjuvant (CFA) and pertussis toxin (PTX) on brain endothelial cells and elucidated the relationships among NADPH oxidase, MMP-9, ICAM-1, and VCAM-1. These 4 factors significantly increased in MOG35-55+CFA+PTX-exposed endothelial cells compared with the control cells. NADPH oxidase inhibition using apocynin reduced MMP-9 activity, ICAM-1, and VCAM-1. MMP-9 inhibitor I decreased expression of ICAM-1 and VCAM-1, and both anti-ICAM-1 and anti-VCAM-1 inhibited MMP-9 activity. Inhibitions of MMP-9, ICAM-1, and VCAM-1 did not change NADPH oxidase activity. Although inhibition of these 4 factors decreased BBB permeability in cells, inhibition of NADPH oxidase exhibited the highest decrease among these. NADPH oxidase directly influenced MMP-9, ICAM-1, and VCAM-1, but not vice versa. MMP-9 and the cell adhesion molecules reversibly affected each other. In conclusion, NADPH oxidase-derived superoxide elevated expression of MMP-9, ICAM-1, and VCAM-1, and these interactions can finally result in increases of BBB permeability in MOG35-55+CFA+PTX-exposed endothelial cells.
Insights
Myelin oligodendrocyte glycoprotein peptide fragment 35-55 (MOG35-55) exposure increases blood-brain barrier permeability. NADPH oxidase activity drives this increase by upregulating MMP-9, ICAM-1, and VCAM-1 in brain endothelial cells.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Multiple sclerosis (MS) involves blood-brain barrier (BBB) disruption.
- Myelin oligodendrocyte glycoprotein peptide fragment 35-55 (MOG35-55) is a key autoantigen in experimental autoimmune encephalomyelitis (EAE), an MS model.
- In vitro BBB models using MOG35-55 are scarce, limiting mechanistic studies.
Purpose of the Study:
- To investigate the effects of MOG35-55 exposure on brain endothelial cells in vitro.
- To elucidate the roles of NADPH oxidase, MMP-9, ICAM-1, and VCAM-1 in MOG35-55-induced BBB changes.
- To determine the causal relationships among these factors and BBB permeability.
Main Methods:
- Establishment of an in vitro BBB model using MOG35-55, complete Freund's adjuvant (CFA), and pertussis toxin (PTX).
- Assessment of NADPH oxidase activity, MMP-9 levels, and ICAM-1/VCAM-1 expression.
- Pharmacological inhibition of NADPH oxidase, MMP-9, ICAM-1, and VCAM-1 to evaluate their impact on BBB permeability.
Main Results:
- MOG35-55+CFA+PTX exposure significantly increased NADPH oxidase, MMP-9, ICAM-1, and VCAM-1 in endothelial cells.
- NADPH oxidase inhibition most effectively reduced BBB permeability.
- NADPH oxidase directly influenced MMP-9, ICAM-1, and VCAM-1, while these factors showed reversible interactions.
Conclusions:
- NADPH oxidase-derived superoxide is a critical upstream regulator of MMP-9, ICAM-1, and VCAM-1 in MOG35-55-induced BBB dysfunction.
- Targeting NADPH oxidase presents a promising therapeutic strategy for mitigating BBB breakdown in MS models.
