Related Experiment Videos
Binding of an anti D dimer monoclonal antibody to endotoxin-activated monocytes. Demonstration by immunogold-silver
S Chollet-Martin1, V Ollivier, J Amiral
1Service d'Immunologie et d'Hématologie Biologiques, CHU X. Bichat, Paris, France.
Abstract:
Blood monocytes exposed to a variety of stimuli, of which endotoxin is a potent one, produce and express on their membrane a procoagulant activity (PCA) which can trigger the formation of pericellular fibrin. We have developed an immunogold silver method using a monoclonal anti-D dimer antibody to detect the presence of crosslinked fibrin in derivatives on the monocyte membrane. The in vitro PCA of endotoxin-stimulated monocyte was shown to correlate significantly with the number of D dimer-positive monocytes. We suggest that this method could be used to identify activated monocytes expressing increased PCA.
Insights
Activated monocytes produce procoagulant activity (PCA) that forms fibrin. A new immunogold silver method using anti-D dimer antibodies detects this fibrin on monocytes, correlating with PCA levels.
Area of Science:
- Hematology
- Immunology
- Biochemistry
Background:
- Blood monocytes are key immune cells involved in coagulation.
- Endotoxin stimulation can induce procoagulant activity (PCA) on monocyte membranes.
- PCA triggers the formation of pericellular fibrin, a marker of activation.
Purpose of the Study:
- To develop a method for detecting crosslinked fibrin on activated monocytes.
- To correlate the presence of fibrin with monocyte procoagulant activity.
- To establish a tool for identifying monocytes with enhanced PCA.
Main Methods:
- Development of an immunogold silver staining technique.
- Utilizing a monoclonal anti-D dimer antibody for specific fibrin detection.
- Quantifying D dimer-positive monocytes in vitro after endotoxin stimulation.
Main Results:
- The immunogold silver method successfully detected crosslinked fibrin on monocyte membranes.
- A significant correlation was observed between in vitro PCA levels and the number of D dimer-positive monocytes.
- The method provides a quantifiable measure of monocyte activation.
Conclusions:
- The developed immunogold silver method is effective for identifying activated monocytes.
- This technique can be used to assess monocyte-derived procoagulant activity.
- This approach offers a novel way to study monocyte-mediated thrombosis and inflammation.