Platelet Microparticles Promote Platelet Interaction With Subendothelial Matrix in a Glycoprotein IIb/IIIa–Dependent Mechanism

Merten, Michael; Pakala, Rajbabu; Thiagarajan, Perumal; Benedict, Claude R.

doi:10.1161/01.cir.99.19.2577

Cited by 139 publications

(101 citation statements)

References 39 publications

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“…After a 60-minute incubation, the bound protein was quantified by peroxidase-labeled monoclonal anti-biotin antibody, with O-phenylenediamine as a substrate, and optical density was measured in an ELISA reader (MR 5000, Dynatech), as previously described. 11 Nonspecific binding of biotin-labeled MCSP, determined in the presence of a 80-fold molar excess of unlabeled MCSP, was subtracted from total binding to obtain specific binding. Nonspecific binding was 11% of total binding.…”

Section: Characterization Of Recombinant Malaria Circumsporozoite Promentioning

confidence: 99%

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Role for Sulfatides in Platelet Aggregation

Merten

Thiagarajan

2001

Circulation

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Background-Sulfatides are sulfated glycosphingolipids present on the surface of oligodendrocytes, renal tubular cells, and certain tumor cells. They appear to be involved in nerve conduction and cell adhesion, but their precise physiological function is not known. Methods and Results-Here, we show a novel role for sulfatides as a major ligand for P-selectin in platelet adhesion and aggregation. Sulfatides are expressed on the platelet surface, and platelets expressing sulfatides adhere to P-selectin. Both sulfatide micelles and sulfatide-binding recombinant malaria circumsporozoite protein (MCSP) inhibit this adhesion. In parallel, platelets and CHO cells expressing P-selectin adhere to sulfatides, and anti-P-selectin antibodies inhibit this adhesion. Furthermore, both anti-P-selectin antibodies and sulfatide antagonist MCSP significantly reverse platelet aggregation induced by ADP, collagen, or thrombin receptor-activating peptide, suggesting that sulfatide-P-selectin interactions are necessary for the formation of stable platelet aggregates. Conclusions-These results show that sulfatide interactions with P-selectin are important in platelet adhesion and platelet aggregation. The sulfatide interactions with P-selectin stabilize platelet aggregates, representing a new mechanism of platelet aggregation that may play a significant role in hemostasis and thrombosis. (Circulation. 2001;104:2955-2960.)

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Section: Characterization Of Recombinant Malaria Circumsporozoite Promentioning

confidence: 99%

“…11 Wells of 96-well microtiter plates were coated with glycolipids or soluble recombinant human P-selectin and blocked as described above. Platelets (1.25ϫ10 6 to 5ϫ10 6 /well) were added in HEPES-buffered saline containing 1% BSA and 1 mmol/L CaCl 2 .…”

Section: Platelet Adhesion Assaysmentioning

confidence: 99%

Role for Sulfatides in Platelet Aggregation

Merten

Thiagarajan

2001

Circulation

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“…29 No specific staining was detected in early or late lesions, indicating that platelet deposition and granule release are unlikely to account for the observed accumulation of LTBP1.…”

Section: Ltbp1 Localization In Coronary Atherosclerosismentioning

confidence: 95%

Localization of Latent Transforming Growth Factor-.BETA. Binding Protein-1 in Human Coronary Atherosclerotic Plaques

Öklü

Hesketh

Wicky

et al. 2011

Circ J

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Background: Transforming growth factor-β (TGFβ) and its receptors have been detected by immunohistochemistry in the normal vessel wall and in atherosclerotic lesions of human coronary arteries. However, TGFβ is normally secreted as an inactive complex associated with a latent TGFβ-binding protein (LTBP). Therefore, detection of TGFβ antigen only in the arterial wall does not imply the activated form of the growth factor. Methods and Results:In situ hybridization and immunohistochemistry demonstrated LTBP1 mRNA and protein expression throughout the media and intima of early coronary artery lesions, with the highest levels of protein at the luminal surface. In advanced lesions, LTBP1 mRNA and protein were detected mainly in regions of high cell density, such as the fibrous cap. Conclusions:Assays of the TGFβ signalling pathway will be required to determine the activity associated with TGFβ antigen in the vessel wall. (Circ J 2011; 75: 196 - 200)

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“…During our initial review of the literature, we found a range of centrifugation conditions used to pellet microparticles: 10,000 Â g for 10 minutes [2], 14,000 Â g for 2 minutes [3], through 100,000 Â g for 1 hour [4,5]. Unable to find a consensus, we chose to use 13,000 Â g for 2 minutes to sediment microparticles for two reasons.…”

Section: Dear Editormentioning

confidence: 99%