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Thromb Haemost 2008; 99(03): 552-557
DOI: 10.1160/TH07-05-0351
Platelets and Blood Cells
Schattauer GmbH

Effect of an anti-sulfatide single-chain antibody probe on platelet function

Prasenjit Guchhait
1   Thrombosis Research Section, Department of Medicine, Baylor College of Medicine, Houston, Texas, USA
,
Corie N. Shrimpton
1   Thrombosis Research Section, Department of Medicine, Baylor College of Medicine, Houston, Texas, USA
,
Koichi Honke
2   Department of Biochemistry, Kochi University Medical School, Kochi, Japan
,
Rolando E. Rumbaut
3   Department of Pediatrics, Baylor College of Medicine, Houston, Texas, USA
,
Jose A. Lopez
4   Research Division, Puget Sound Blood Center, Seattle, Washington, USA
,
Perumal Thiagarajan
1   Thrombosis Research Section, Department of Medicine, Baylor College of Medicine, Houston, Texas, USA
3   Department of Pediatrics, Baylor College of Medicine, Houston, Texas, USA
› Author Affiliations Financial support: This study was supported by a Beginning Grant-In-Aid to P.G. by the American Heart Association (0565044Y) and a R01 grant to R.E.R by NIH/NHLBIHL (079368).
Further Information

Publication History

Received: 15 May 2007

Accepted after major revision: 08 January 2008

Publication Date:
07 December 2017 (online)

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Summary

Sulfatide (galactocylceramide-3'-sulfate), a cell surface glycosphingolipid interacts with several cell adhesion molecules including fibrinogen, von Willebrand factor (VWF), P-selectin, thrombospondin (TSP) and laminin, which are involved in haemostasis.We have used a sulfatide-specific single-chain fragment variable (scFv) antibody probe PA38 and sulfatide-deficient mice to investigate the role of membrane sulfatide in platelet function. PA38 bound to platelets and binding increased following platelet activation. Sulfatide was localized as a large cluster towards the center of the platelet surface when examined in a confocal microscope. PA38 (20 μg/ml) inhibited the adhesion of activated platelets to fibrinogen,VWF, P-selectin,TSP1 and laminin by 30%, 30%,75%,20% and 35%,respectively,compared to a control scFv (p<0.05). Furthermore, PA38 inhibited collagen, ADP, thrombin and ristocetin-induced platelet aggregation in PRP by 25%, 30%, 18% and 20%, respectively, compared to the control scFv (p<0.05). In a PFA-100 platelet function assay, PA38 prolonged the occlusion time by 25% (p<0.05).Under flow PA38 decreased the thrombus formation on collagen by 31%, (p<0.01). Sulfatidedeficient mice displayed an extended lag-phase in collagen-induced platelet aggregation compared to wild type (p<0.05), though in-vivo haemostasis did not differ significantly.Thus, this study provides new evidence for a role for membrane sulfatide in platelet function.

Keywords

Sulfatide - anti-sulfatide scFv - platelet function
 

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