Therapeutic TNF Inhibitors can Differentially Stabilize Trimeric TNF by Inhibiting Monomer Exchange
| Autor: | Pleuni Ooijevaar-de Heer, Theo Rispens, Gertjan Wolbink, Karin A van Schie, Lisanne Dijk, Simone Kruithof |
|---|---|
| Přispěvatelé: | Landsteiner Laboratory |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Protein subunit medicine.medical_treatment Certolizumab Article Etanercept 03 medical and health sciences 0302 clinical medicine Adalimumab medicine Fluorescence Resonance Energy Transfer Humans Multidisciplinary Chemistry Protein Stability Tumor Necrosis Factor-alpha Antibodies Monoclonal Surface Plasmon Resonance In vitro Golimumab Infliximab Kinetics 030104 developmental biology Cytokine 030220 oncology & carcinogenesis Immunology Biophysics Chromatography Gel Tumor necrosis factor alpha Protein Multimerization medicine.drug |
| Zdroj: | Scientific Reports Scientific reports, 6. Nature Publishing Group |
| ISSN: | 2045-2322 |
| Popis: | Tumor necrosis factor (TNF) is a homotrimeric cytokine that is a key mediator of inflammation. It is unstable at physiological concentrations and slowly converts into an inactive form. Here, we investigated the mechanism of this process by using a Förster resonance energy transfer (FRET) assay that allowed monitoring of monomeric subunit exchange in time. We observed continuous exchange of monomeric subunits even at concentrations of TNF high enough to maintain its bioactivity. The kinetics of this process closely corresponds with the appearance of monomeric subunits and disappearance of trimeric TNF in time at ng/ml concentrations as monitored by high-performance size-exclusion chromatography (HP-SEC). Furthermore, of the five therapeutic TNF inhibitors that are currently used in the clinic, three (adalimumab, infliximab, etanercept) were found to completely inhibit the monomer exchange reaction and stabilize TNF trimers, whereas golimumab and certolizumab could not prevent monomer exchange, but did slow down the exchange process. These differences were not correlated with the affinities of the TNF inhibitors, measured with both surface plasmon resonance (SPR) and in fluid phase using fluorescence-assisted HP-SEC. The stabilizing effect of these TNF inhibitors might result in prolonged residual TNF bioactivity under conditions of incomplete blocking, as observed in vitro for adalimumab. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|