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Updated: Jan 12, 2026
Author Spotlight: Advancing the Analysis of Plasma Extracellular Vesicle Proteome for Cardiovascular Biomarker Studies
Published on: January 31, 2025
Comparative analysis of plasma proteolytic profiles in multiple sclerosis patients with and without a history of
Larysa Kot1, Olena Strubchevska2, Tetiana Halenova1
1Taras Shevchenko National University of Kyiv, Volodymyrska Str. 64/13, Kyiv, 01601, Ukraine.
Background:
Multiple sclerosis (MS) is a neurodegenerative disorder of the central nervous system characterized by abnormal proteolytic activity-particularly involving serine proteases-that contributes to neuroinflammation and tissue damage. The COVID-19 pandemic has raised concerns about long-term immune and metabolic disturbances that may exacerbate MS progression. This study compared plasma proteolytic profiles in MS patients with and without prior COVID-19, focusing on serine protease composition, molecular organization, and middle-mass molecule (MMM) levels as markers of proteolytic imbalance and systemic metabolic changes.
Methods:
A total of 127 participants aged 35-45 years were recruited from the University Clinic, Bogomolets National Medical University (Kyiv, Ukraine). Ninety-seven had MS and were divided into two groups: MS with COVID-19 (n = 41; confirmed SARS-CoV-2 infection 3-6 months prior) and MS without COVID-19 (n = 56). Thirty healthy volunteers served as controls. Proteolytic activity was assessed via fibrinogen- and gelatin-substrate zymography. Serine protease fractions were isolated by affinity chromatography and quantified by the Bradford assay. Protein composition was analyzed by SDS-PAGE under nonreducing and reducing conditions.
Results:
In MS patients, zymography revealed altered proteolytic profiles with a shift toward high-molecular-weight fibrinogenolytic complexes and redistribution of gelatinolytic activity. These changes were more pronounced in those with prior COVID-19. Total plasma serine protease levels were significantly higher in MS patients, particularly in the post-COVID-19 group, which also showed notable alterations in molecular composition and activity patterns. Moreover, MMM peptide fractions related to nonaromatic and aromatic compounds were markedly reduced, especially in post-COVID-19 MS patients, indicating systemic metabolic disturbances.
Conclusions:
MS is associated with persistent quantitative and qualitative changes in plasma proteolytic activity, further amplified by prior SARS-CoV-2 infection. The combination of elevated serine protease levels, high-molecular-weight complex formation, and depletion of specific MMM fractions suggests a sustained imbalance in protease homeostasis. These alterations may contribute to chronic inflammation, endothelial dysfunction, and extracellular matrix remodeling. Plasma protease profiling could serve as a diagnostic and therapeutic tool in MS, particularly for monitoring post-COVID-19 complications.
