TY - JOUR
T1 - The active form of MMP-3 is a marker of synovial inflammation and cartilage turnover in inflammatory joint diseases
AU - Sun, Shu
AU - Bay-Jensen, Anne-Christine
AU - Karsdal, Morten A
AU - Siebuhr, Anne Sofie
AU - Zheng, Qinlong
AU - Maksymowych, Walter P
AU - Christiansen, Thorbjørn G
AU - Henriksen, Kim
PY - 2014
Y1 - 2014
N2 - BACKGROUND: Matrix metalloproteinase-3 (MMP-3) plays an important role in the pathology of rheumatoid arthritis (RA) and ankylosing spondylitis (AS). Measurement of active MMP-3 in clinical samples could provide information about progression of rheumatoid diseases, and potentially response to treatment. Hence, we aimed to develop a sensitive assay specifically measuring the active form of MMP-3 (act-MMP-3) both in ex vivo models and in human sera.METHODS: A monoclonal antibody against the first 6 amino acids of act-MMP-3 was developed, and the specificity was carefully tested by comparing total and active MMP-3. A technically robust act-MMP-3 ELISA was produced. For biological validation, human synovial membrane and human cartilage explant (HEX) culture models were measured and compared by ELISA and immunoblots. For clinical relevance, the serum levels of act-MMP-3 in AS and RA patients before and after anti-TNF-α treatment were evaluated.RESULTS: A highly specific and technically robust ELISA detecting act-MMP-3 in serum was developed. The lower limit of detection was 33.7 pg/mL. The dilution and spiking recovery of human serum was within 100 ± 20%. The average intra- and inter-assay variations were 3.1% and 13.5% respectively.High levels of act-MMP-3 expression were observed in human synovial membrane culture and oncostatin M and TNF-α stimulated human cartilage. In a cross-sectional study of both AS and RA patients, serum act-MMP-3 level was correlated with C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR). In addition, in patients receiving anti-TNF-α treatment, the serum level of act-MMP-3 was significantly reduced compared to baseline level reflecting the anti-inflammatory effects of the treatment.CONCLUSION: We have successfully developed an assay measuring act-MMP-3 in human serum showing correlation to inflammatory markers. Further studies are required to clarify, whether act-MMP-3 can serve as a predictive marker for outcome in chronic rheumatoid disorders.
AB - BACKGROUND: Matrix metalloproteinase-3 (MMP-3) plays an important role in the pathology of rheumatoid arthritis (RA) and ankylosing spondylitis (AS). Measurement of active MMP-3 in clinical samples could provide information about progression of rheumatoid diseases, and potentially response to treatment. Hence, we aimed to develop a sensitive assay specifically measuring the active form of MMP-3 (act-MMP-3) both in ex vivo models and in human sera.METHODS: A monoclonal antibody against the first 6 amino acids of act-MMP-3 was developed, and the specificity was carefully tested by comparing total and active MMP-3. A technically robust act-MMP-3 ELISA was produced. For biological validation, human synovial membrane and human cartilage explant (HEX) culture models were measured and compared by ELISA and immunoblots. For clinical relevance, the serum levels of act-MMP-3 in AS and RA patients before and after anti-TNF-α treatment were evaluated.RESULTS: A highly specific and technically robust ELISA detecting act-MMP-3 in serum was developed. The lower limit of detection was 33.7 pg/mL. The dilution and spiking recovery of human serum was within 100 ± 20%. The average intra- and inter-assay variations were 3.1% and 13.5% respectively.High levels of act-MMP-3 expression were observed in human synovial membrane culture and oncostatin M and TNF-α stimulated human cartilage. In a cross-sectional study of both AS and RA patients, serum act-MMP-3 level was correlated with C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR). In addition, in patients receiving anti-TNF-α treatment, the serum level of act-MMP-3 was significantly reduced compared to baseline level reflecting the anti-inflammatory effects of the treatment.CONCLUSION: We have successfully developed an assay measuring act-MMP-3 in human serum showing correlation to inflammatory markers. Further studies are required to clarify, whether act-MMP-3 can serve as a predictive marker for outcome in chronic rheumatoid disorders.
KW - Amino Acid Sequence
KW - Animals
KW - Antibodies, Monoclonal
KW - Antirheumatic Agents
KW - Arthritis, Rheumatoid
KW - Biological Markers
KW - C-Reactive Protein
KW - Cartilage
KW - Enzyme Activation
KW - Enzyme-Linked Immunosorbent Assay
KW - Female
KW - Humans
KW - Inflammation
KW - Male
KW - Matrix Metalloproteinase 3
KW - Mice
KW - Molecular Sequence Data
KW - Oncostatin M
KW - Organ Culture Techniques
KW - Osteoarthritis, Knee
KW - Reproducibility of Results
KW - Sensitivity and Specificity
KW - Severity of Illness Index
KW - Specific Pathogen-Free Organisms
KW - Spondylitis, Ankylosing
KW - Synovial Membrane
KW - Treatment Outcome
KW - Tumor Necrosis Factor-alpha
U2 - 10.1186/1471-2474-15-93
DO - 10.1186/1471-2474-15-93
M3 - Journal article
C2 - 24641725
SN - 1471-2474
VL - 15
SP - 93
JO - B M C Musculoskeletal Disorders
JF - B M C Musculoskeletal Disorders
ER -