Flow cytometric assessment of agonist‐induced <scp>P</scp>‐selectin expression as a measure of platelet quality in stored platelet concentrates

Middelburg, Rutger A.; Roest, Mark; Ham, Jannemieke; Coccoris, Miriam; Zwaginga, Jaap Jan; Meer, Pieter F. van der

doi:10.1111/trf.12001

Cited by 38 publications

(54 citation statements)

References 14 publications

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“…We were able to show the decrease in ThromboLUX scores as a consequence of the degradation of apheresis platelets during storage as well as an additional effect with the Mirasol pathogen reduction treatment. Our results are in agreement with other studies [26].…”

Section: Comparison Of Thrombolux Results To Microscopysupporting

confidence: 83%

“…The presence of microparticles might therefore be an indicator of diminished platelet quality. Changes of platelet characteristics in concentrates have been reported as a function of storage time and treatments such as pathogen reduction [26]. We were able to show the decrease in ThromboLUX scores as a consequence of the degradation of apheresis platelets during storage as well as an additional effect with the Mirasol pathogen reduction treatment.…”

Section: Comparison Of Thrombolux Results To Microscopysupporting

confidence: 59%

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Characterization of Platelet Concentrates Using Dynamic Light Scattering

Labrie¹,

Marshall

Bedi

et al. 2013

Transfus Med Hemother

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Background: Each year, millions of platelet transfusions save the lives of cancer patients and patients with bleeding complications. However, between 10 and 30% of all platelet transfusions are clinically ineffective as measured by corrected count increments, but no test is currently used to identify and avoid these transfusions. ThromboLUX® is the first platelet test intended to routinely characterize platelet concentrates prior to transfusion. Methods: ThromboLUX is a non-invasive, optical test utilizing dynamic light scattering to characterize a platelet sample by the relative quantity of platelets, microparticles, and other particles present in the sample. ThromboLUX also determines the response of platelets to temperature changes. From this information the ThromboLUX score is calculated. Increasing scores indicate increasing numbers of discoid platelets and fewer microparticles. ThromboLUX uses calibrated polystyrene beads as a quality control standard, and accurately measures the size of the beads at multiple temperatures. Results: Results from apheresis concentrates showed that ThromboLUX can determine the microparticle content in unmodified samples of platelet concentrates which correlates well with the enumeration by flow cytometry. ThromboLUX detection of microparticles and microaggregates was confirmed by microscopy. Conclusion: ThromboLUX provides a comprehensive and novel analysis of platelet samples and has potential as a non-invasive routine test to characterize platelet products to identify and prevent ineffective transfusions.

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Section: Comparison Of Thrombolux Results To Microscopysupporting

confidence: 83%

Section: Comparison Of Thrombolux Results To Microscopysupporting

confidence: 59%

Characterization of Platelet Concentrates Using Dynamic Light Scattering

Labrie¹,

Marshall

Bedi

et al. 2013

Transfus Med Hemother

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“…Platelet function tests were performed by adding 5 μL of whole blood per test as previously described. 4 In short, concentration series of collagen-related peptide (CRP) and thrombin receptor activating peptide (TRAP), in the presence of FITC-labelled mouse anti-human CD61 antibodies (Becton Dickinson, Breda, The Netherlands, diluted 1:25) and PE-labelled mouse anti-human P-selectin antibodies (Becton Dickinson, Breda, The Netherlands, diluted 1:25) were prepared in HEPES buffered saline (HBS). Agonist concentration series were prepared by serial dilution in 7 steps of 1:4 with starting concentrations for CRP 2.5 μg/mL, and for TRAP 625 nM.…”

Section: Methodsmentioning

confidence: 99%

“…relative MFI in the PE channel), as previously suggested. 4,5 Correction for platelet count, as a potential confounder, was performed in four categories based on the quartiles of platelet counts (<32, 32-72, 73-132, >132 × 10 9 platelets/L), using the Mantel-Haenszel method. 6 The observed concentration response profiles for platelets from ITP patients were classified as 'increased activity', 'normal activity', or 'decreased activity', compared to the observed profiles in healthy volunteers.…”

Section: Methodsmentioning

confidence: 99%

Platelet function in adult ITP patients can be either increased or decreased, compared to healthy controls, and is associated with bleeding risk

et al. 2016

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Objectives: To test whether, together with platelet count, platelet activity could be an important predictor of bleeding risk in immune thrombocytopenia (ITP) patients.Methods: Platelet activity was tested by flow cytometric measurement of agonist induced P-selectin expression and compared between 23 adult ITP patients and 22 healthy volunteers. Results: Platelet activity could be either increased or decreased in ITP patients, compared to healthy volunteers. In the lowest platelet count category, normal to low platelet activity was associated with the biggest increase in bleeding risk. Risk difference 80% (95% confidence interval: 45-115%) for <32 × 10 9 platelets/L. For higher platelet counts, there was no association of platelet activity with bleeding risk. Discussion: Increased platelet activity was associated with decreased bleeding risk, but only in patients with low platelet counts. Conclusion: Platelet activity can be a predictor of bleeding risk in ITP patients with low platelet counts.

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“…The main advantage of pac-t-UB is that all activation pathways can separately be investigated in a single experiment using only a minimal amount of unprocessed blood. The pac-t-UB has been validated by our laboratory in several studies on the relation between platelet function and (secondary) cardiovascular disease incidents as well as on the storage of untreated versus Mirasol-treated platelets in plasma [5]. Applications for monitoring P2Y 12 and aspirin treatment and diagnosis for (un) known bleeding disorders are ongoing.…”

Section: Introductionmentioning

confidence: 99%

Platelet Activation Test in Unprocessed Blood (Pac-t-UB) to Monitor Platelet Concentrates and Whole Blood of Thrombocytopenic Patients

Roest¹,

Holten²,

Fleurke³

et al. 2013

Transfus Med Hemother

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Background: Platelet concentrate transfusion is the standard treatment for hemato-oncology patients to compensate for thrombocytopenia. We have developed a novel platelet activation test in anticoagulated unprocessed blood (pac-t-UB) to determine platelet function in platelet concentrates and in blood of thrombocytopenic patients. Methods: We have measured platelet activity in a platelet concentrate and in anticoagulated unprocessed blood of a post-transfusion thrombocytopenic patient. Results: Our data show time-dependent platelet activation by GPVI agonist (collagen related peptide; CRP), PAR-1 agonist (SFLLRN), P2Y₁₂ agonist (ADP), and thromboxane receptor agonist (U46619) in a platelet concentrate. Furthermore, pac-t-UB showed time-dependent platelet activation in unprocessed blood of a post-transfusion patient with thrombocytopenia. Testing platelet function by different agonists in relation to storage show that 3-day-old platelet concentrates are still reactive to the studied agonists. This reactivity rapidly drops for each agonists during longer storage. Discussion: Pac-t-UB is a novel tool to estimate platelet function by different agonists in platelet concentrates and in unprocessed blood of thrombocytopenic patients. In the near future, we will validate whether pac-t-UB is an adequate test to monitor the quality of platelet concentrates and whether pac-t-UB predicts the bleeding risk of transfused thrombocytopenic patients.

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Flow cytometric assessment of agonist‐induced P‐selectin expression as a measure of platelet quality in stored platelet concentrates

Abstract: Our results suggest flow cytometric measurement of agonist-induced P-selectin expression can measure PLT quality decline over the entire range encountered during 10-day storage of both standard PLTs and Mirasol-treated PLTs in plasma.

Cited by 38 publications

References 14 publications

Characterization of Platelet Concentrates Using Dynamic Light Scattering

Characterization of Platelet Concentrates Using Dynamic Light Scattering

Platelet function in adult ITP patients can be either increased or decreased, compared to healthy controls, and is associated with bleeding risk

Platelet Activation Test in Unprocessed Blood (Pac-t-UB) to Monitor Platelet Concentrates and Whole Blood of Thrombocytopenic Patients

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