ABSTRACT
Mannose binding lectin (MBL) mediated complement pathway is an important constituent of innate immune response in several infections including neuroinflammatory and neurodegenerative diseases. Although there are Enzyme-Linked Immunosorbent Assays (ELISAs) for estimating MBL, MBL-associated serine protease-2 (MASP-2) and functional MBL-MASP-2 (fMBL) proteins for the plasma, serum and cell supernatants there are no established methods for their estimation in the cerebrospinal fluid (CSF). We developed sensitive ELISAs for the detection of MBL, fMBL and MASP-2 in the CSF. First, we adapted standard ELISAs for the detection of these proteins in the CSF. Second, we used a biotinyl-tyramide based horseradish peroxidase (HRP) signal amplification for the sensitive detection of these proteins in the CSF. In summary, using modified ELISA and biotinyl-tyramide based HRP signal amplification, we successfully detected MBL, fMBL and MASP-2 proteins in the CSF samples with high sensitivity and reproducibility.
Subject(s)
Enzyme-Linked Immunosorbent Assay/methods , Mannose-Binding Lectin/cerebrospinal fluid , Mannose-Binding Protein-Associated Serine Proteases/cerebrospinal fluid , Humans , Sensitivity and SpecificityABSTRACT
Role of mannose binding lectin (MBL) complement activation pathway, an arm of innate immunity in multiple sclerosis (MS) was evaluated by analyzing the expression of MBL, MBL-associated serine protease-2 (MASP-2), and functional MBL/MASP-2 mediated C4 cleavage (fMBL) in 87 plasma and cerebrospinal fluid (CSF) samples from MS patients and non-MS controls. Median fMBL and MASP-2 plasma levels were higher in MS vs. non-MS cases. These associations remained in an analysis of subtypes of MS disease. These findings suggest a potential activation of MBL complement pathway in MS that may possibly alter the risk or progression of MS disease.
