Profiling and Identification of Cerebrospinal Fluid Proteins in a Rat EAE Model of Multiple Sclerosis

Abstract

The experimental autoimmune encephalomyelitis (EAE) model resembles certain aspects of multiple sclerosis (MScl), with common features such as motor dysfunction, axonal degradation, and infiltration of T-cells. We studied the cerebrospinal fluid (CSF) proteome in the EAE rat model to identify proteomic changes relevant for MScl disease pathology. EAE was induced in male Lewis rats by injection of myelin basic protein (MBP) together with complete Freund's adjuvant (CFA). An inflammatory control group was injected with CFA alone, and a nontreated group served as healthy control. CSF was collected at day 10 and 14 after immunization and analyzed by bottom-up proteomics on Orbitrap LC-MS and QTOF LC-MS platforms in two independent laboratories. By combining results, 44 proteins were discovered to be significantly increased in EAE animals compared to both control groups, 25 of which have not been mentioned in relation to the EAE model before. Lysozyme C1, fetuin B, T-kininogen, serum paraoxonase/arylesterase 1, glutathione peroxidase 3, complement C3, and afamin are among the proteins significantly elevated in this rat EAE model. Two proteins, afamin and complement C3, were validated in an independent sample set using quantitative selected reaction monitoring mass spectrometry. The molecular weights of the identified differentially abundant proteins indicated an increased transport across the blood-brain barrier (BBB) at the peak of the disease, caused by an increase in BBB permeability.

Discovery of Novel Disease-specific and Membrane-associated Candidate Markers in a Mouse Model of Multiple Sclerosis

“…Biological Significance of the Putative Disease Markers Detected in the Spinal Cord-Elevated levels of known protein members of the coagulation cascade such as fibrinogen (alpha, beta, and gamma isoforms) and kininogen-1, were identified by both spectral counting analysis and MS 1 -based quantification in the diseased spinal cord, observations that have previously been made by proteomic analyses of chronic active plaques isolated from post-mortem MScl brain tissue (12) and CSF from the MBP-EAE model (28). In concordance with other studies, we also identified increased levels of vitamin-d-binding protein (Vdbp), several protein members of the immunoproteasome, and multiple isoforms of immunoglobulin G (IgG) within the diseased spinal cord by both label-free quantification methods.…”

“…Rosenling and colleagues recently identified 44 differentially abundant proteins within the CSF of a rodent EAE model of which only two were validated by MRM (28). In this study, three to four unique peptides served as protein surrogates in the MRM-based validation of candidate disease markers with the best four transitions per peptide monitored.…”

Selenium, selenoproteins and neurodegenerative diseases

“…170,171 In a proteomic study, among a set of 44 proteins that discriminated experimental autoimmune encephalomyelitis (EAE) rats from control groups, GPx3 was increased in EAE animals. 172 Selenium levels tend to decrease in MS, as observed in a cohorts of Iranian 173 and Polish patients. 174 Complementing the suggestive data regarding a role of Se in MS pathogenesis, Chanaday et al 66 observed that treatment with diphenyl diselenide to the EAE animal model reduced signs of the disease phenotype, partly by preventing the reduction in body weight that accompanies the disease progression and decreased the time and the severity of symptoms.…”

Afamin promotes glucose metabolism in papillary thyroid carcinoma

“…AFM has been shown to be a specific binding protein for vitamin E (Lichenstein et al, 1994). Also, it has been reported that AFM had the effect of neuroprotection (Heiser et al, 2002) and possible marker for various neurological pathologies including Alzheimer's disease and multiple sclerosis (Ringman et al, 2012;Rosenling et al, 2012). Diseases including pregnancy complications (Hubalek et al, 2014), metabolic syndrome (diabetes mellitus, obesity and hypertension), polycystic ovary syndrome (Koninger et al, 2014) have been showed associated with AFM as well.…”