The association of the <scp>T</scp>hr715<scp>P</scp>ro <scp>P</scp>‐selectin genotype with levels of <scp>P</scp>‐selectin in platelet concentrates

Jilma‐Stohlawetz, Petra; Mannhalter, Christine; Kaider, Alexandra; Waidacher, Theresa; Jilma, Bernd; Panzer, Simon

doi:10.1111/vox.12175

Cited by 4 publications

(8 citation statements)

References 34 publications

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“…The genetic distribution of the SELP Pro715 allele was in line with our previous observations, and was similar in patients and in the group of healthy controls (11,29). Also, the distribution of the SELPLG Ile62 allele was similar in patients and healthy controls.…”

Section: Discussionsupporting

confidence: 91%

“…While confirming the association of sP-selectin levels with the SELP Thr715Pro polymorphism, our current data extend these previous findings by showing that also levels of pP-selectin are genotype-associated. This is supported by our results obtained with platelet concentrates to limit the contribution of P-selectin from endothelial Weibel-Palade bodies, where we could demonstrate that total P-selectin from platelets (comprising sP-selectin together with pP-selectin and P-selectin stored in the alpha granules), is SELP Thr715Pro-associated (11). Likewise, an association between pP-selectin and sP-selectin appeared likely.…”

Section: Discussionsupporting

confidence: 84%

“…P-selectin is rapidly shed from the platelet surface and is then found in its soluble form in the plasma (sP-selectin). SP-selectin derives mainly from platelets (11), and similar to pP-selectin can bind to PSGL-1 and activate leukocytes (12,13). Experimental evidence shows that sP-selectin has procoagulant activity (14).…”

Section: Introductionmentioning

confidence: 99%

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Impact of variables of the P-selectin – P-selectin glycoprotein ligand-1 axis on leukocyte-platelet interactions in cardiovascular disease

Gremmel

Koppensteiner²,

Kaider³

et al. 2015

Thromb Haemost

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The formation of leukocyte-platelet aggregates (LPA), through the P-selectin - P-selectin glycoprotein ligand (PSGL)-1 axis, plays a pivotal role in atherothrombosis. In order to investigate the influence of platelet (pP-selectin) and soluble P-selectin (sP-selectin), and of variations in the genes encoding for P-selectin (SELP) and PSGL-1 (SELPLG) on LPA formation, we assessed monocyte (MPA)- and neutrophil-platelet aggregates (NPA) as well as pP-selectin by flow cytometry in 263 patients undergoing angioplasty and stenting. sP-selectin was determined by ELISA, the SELP Pro715 allele and the SELPLG Ile62 allele were determined by allele specific PCR. The Pro715 allele was significantly associated with lower levels of in vivo pP-selectin and sP-selectin, while agonists´ inducible pP-selectin was not influenced by the Pro715 allele. PP-selectin was significantly associated with MPA and NPA formation. The in vivo formation of MPA and NPA depended to 19 % and 7.4 %, respectively, on in vivo pP-selectin, irrespective of the Pro715 allele and the Ile62 allele carrier status. TRAP-6 inducible MPA and NPA depended to 34 % and 27 %, respectively, on TRAP-6 inducible pP-selectin, but were independent of the Pro715 allele carrier status. Carriers of the Ile62 allele showed a stronger correlation between TRAP-6 inducible pP-selectin and TRAP-6 inducible MPA/NPA than non-carriers. Furthermore, TRAP-6 inducible NPA were higher in Ile62 allele carriers, which suggests higher thrombin sensitivity. In conclusion, our findings point to the significant role of pP-selectin for MPA and NPA formation, while other variables like sP-selectin, the SELP Pro715 allele and the SELPLG Ile62 allele have less influence.

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Section: Discussionsupporting

confidence: 91%

Section: Discussionsupporting

confidence: 84%

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Impact of variables of the P-selectin – P-selectin glycoprotein ligand-1 axis on leukocyte-platelet interactions in cardiovascular disease

Gremmel

Koppensteiner²,

Kaider³

et al. 2015

Thromb Haemost

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“…Taking into account the characteristics of all the methods available, the PCR‐HRM method was optimal choice for our laboratory. PCR‐RFLP, PCR‐SSP, and mutagenically separated PCR are relatively simple and low‐cost methods that do not require special and expensive equipment but are time‐consuming. These methods might be appropriate for laboratories that have fewer samples to be tested, with limited reliability for assessing correct genotypes due to greater error potential.…”

Section: Discussionmentioning

confidence: 99%

“…A limited number of methods for genotyping the NM_003005.3:c.992G>A (c.992G>A), NM_003005.3:c.1918G>T (c.1918G>T), and NM_003005.3:c.2266A>C (c.2266A>C) SELP polymorphisms (historically named S290N, V599L, and T715P, respectively) are available in the literature and include PCR‐restriction fragment length polymorphism (RFLP), polymerase chain reaction (PCR) with sequence‐specific primers (SSP), mutagenically separated PCR, TaqMan technology‐based real‐time PCR, HRM analysis, and sequencing methods …”

Section: Introductionmentioning

confidence: 99%

Development and validation of a rapid method for genotyping three P‐selectin gene polymorphisms based on high resolution melting analysis

Čeri

Pavić

Horvat

et al. 2018

Clinical Laboratory Analysis

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Background High resolution melting (HRM) analysis is one of the newer, reliable, and sensitive genotyping techniques, which offers considerable time and cost savings. P‐selectin is an adhesion molecule that has a role in the initial phases of leukocyte adhesion to stimulated platelets and endothelial cells in inflammation. Multiple polymorphisms in P‐selectin gene (SELP) that affect the protein sequence have been described. The aim of this study was to design, optimize, and validate a simple and rapid in‐house HRM‐based method for genotyping the NM_003005.3:c.992G>A (c.992G>A), NM_003005.3:c.1918G>T (c.1918G>T), and NM_003005.3:c.2266A>C (c.2266A>C) SELP polymorphisms. Methods Initial genotyping of three SELP polymorphisms was performed by applying polymerase chain reaction (PCR) with sequence‐specific primers (SSP), which was used as a reference method for determination of analytical sensitivity. PCR‐HRM was performed with primers for c.2266A>C reported in the literature. Primers for the remaining two polymorphisms were designed using Primer‐BLAST. Precision testing was performed using three samples with different genotypes. For accuracy, analytical sensitivity and specificity testing, 20 wild type, 10 heterozygous, and 10 homozygous samples were chosen per polymorphism. Results were expressed as percentage of concordance with the acceptability criterion ≥95%. Results Agreement of results was 100% for all validation parameters except for analytical sensitivity for c.1918G>T and c.2266A>C, with agreement of 90%. Repeated analysis using both methods revealed an error in initial genotyping and correct genotyping by PCR‐HRM, which was confirmed by Sanger sequencing. Conclusion The validation confirmed PCR‐HRM as a precise, accurate, and specific method for genotyping the c.992G>A, c.1918G>T, and c.2266A>C SELP polymorphisms.

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Platelets in Deep Venous Thrombosis and Pulmonary Embolism

Pabinger

Riedl

Panzer

2017

Platelets in Thrombotic and Non-Thrombotic Disorders

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The association of the Thr715Pro P‐selectin genotype with levels of P‐selectin in platelet concentrates

Abstract: The donors' genotype has only little influence on levels of soluble P-selectin in apheresis platelets suspended in 35% plasma/65% SSP+.

Cited by 4 publications

References 34 publications

Impact of variables of the P-selectin – P-selectin glycoprotein ligand-1 axis on leukocyte-platelet interactions in cardiovascular disease

Impact of variables of the P-selectin – P-selectin glycoprotein ligand-1 axis on leukocyte-platelet interactions in cardiovascular disease

Development and validation of a rapid method for genotyping three P‐selectin gene polymorphisms based on high resolution melting analysis

Platelets in Deep Venous Thrombosis and Pulmonary Embolism

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