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Platelet glycoproteins IIb and IIIa associated with blood monocytes are derived from platelets
Abstract:
Platelet glycoprotein IIb/IIIa (GP IIb/IIIa), the receptor complex for fibrinogen, has been regarded as a megakaryocyte/platelet lineage-restricted antigen. Recently, however, it has been reported that GP IIb/IIIa is expressed in blood monocytes. Studies were performed to establish the origin and immunological characteristics of monocyte-associated glycoproteins IIb and IIIa (GPs IIb and IIIa). Preparations of blood monocytes containing varying platelet-monocyte ratios were metabolically labeled with [35S]methionine with the expectation that any newly synthesized GPs IIb and IIIa would be monocyte-derived, since platelets have only rudimentary protein synthetic apparatuses. Analyses of sodium dodecyl sulfate (SDS) gels of homogenates of cell preparations containing from 200 to 5:1 platelet-monocyte ratios revealed that unlabeled GPs IIb and IIIa were readily immunoisolated using protein A-Sepharose immunobeads. However, fluorographic analyses of the same cell preparations pulse-labeled with [35S]methionine failed to demonstrate synthesis of GP IIb or IIIa. Additionally, no GP IIb or IIIa was detected when immunoisolation was carried out in pure preparations of monocytes containing less than 1:100 platelet-monocyte ratios and SDS acrylamide gels were stained by the sensitive silver stain method. Furthermore, heterologous polyspecific antisera and two monoclonal antibody preparations against GPs IIb and IIIa, which bound to platelets, failed to bind to monocyte membranes. Thus, evidence was presented that indicated that monocytes do not synthesize platelet GPs IIb and IIIa and that detection of these molecules in blood monocyte preparations reflects platelet contamination.
Insights
Monocytes do not synthesize platelet glycoprotein IIb/IIIa (GP IIb/IIIa). Detection of GP IIb/IIIa in monocytes is due to platelet contamination, not monocyte expression.
Area of Science:
- Immunology
- Hematology
- Cell Biology
Background:
- Platelet glycoprotein IIb/IIIa (GP IIb/IIIa) is a fibrinogen receptor.
- GP IIb/IIIa was traditionally considered exclusive to megakaryocytes and platelets.
- Recent findings suggested GP IIb/IIIa expression on blood monocytes.
Purpose of the Study:
- To investigate the origin of monocyte-associated GP IIb/IIIa.
- To determine if monocytes synthesize GP IIb/IIIa.
- To clarify the immunological characteristics of monocyte-associated GPs IIb and IIIa.
Main Methods:
- Metabolic labeling of monocytes with [35S]methionine to detect protein synthesis.
- Immunoisolation of GPs IIb and IIIa using specific antibodies.
- Analysis of protein expression via SDS-PAGE and silver staining.
- Testing antibody binding to monocyte membranes.
Main Results:
- Pulse-labeling experiments failed to demonstrate GP IIb/IIIa synthesis by monocytes.
- GP IIb/IIIa was not detected in highly purified monocyte preparations.
- Antibodies against GP IIb/IIIa did not bind to monocyte membranes.
- Unlabeled GP IIb/IIIa was immunoisolated from monocyte preparations with varying platelet contamination.
Conclusions:
- Monocytes do not synthesize platelet GP IIb/IIIa.
- Observed GP IIb/IIIa in monocyte preparations is attributable to platelet contamination.
- GP IIb/IIIa is not a monocyte-derived antigen.