Assessment of antiplatelet therapy response in pediatric patients following cardiac surgery by microfluidic assay

Kapileshwarkar, Yamini; Gelmini, Lucas; Tseng, Yu Shan; Jackson, Tannoa; Gao, Xiufeng; Richards, Kristen; Dokas, Michelle; Walters, Henry L.; Hines, Patrick C.

doi:10.1016/j.ppedcard.2019.101191

Cited by 2 publications

(2 citation statements)

References 39 publications

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“…Briefly, isolated white blood cell (i-WBC) suspensions were prepared according to a standardized protocol (HetaSep TM , StemCell Technologies). Whole blood samples (1:1 diluted with HBSS buffer) or i-WBC (5x 10 6 cells/mL) were perfused through P-Selectin-coated microfluidic channels at 1 dynes/cm 2 for 10 or 6 minutes respectively, washed to eliminate non-adhering cells, with resultant adhesion quantified manually by an independent trained observer to generate an adhesion index (cells/mm²) using a previously described protocol [25]. For drug treatment conditions, samples were incubated for 5 minutes with crizanlizumab at the final concentrations as required before assessment by the flow adhesion assay.…”

Section: Methodsmentioning

confidence: 99%

Possible Role of P-selectin Adhesion in Long-COVID: A Comparative Analysis of a Long-COVID Case Versus an Asymptomatic Post-COVID Case

Tarasev

Gao

Mota

et al. 2022

Preprint

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Background: Long-term outcomes of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are now recognized as an emerging public health challenge - a condition termed Long-COVID. The pathophysiology of Long-COVID remains to be established. Functional P-selectin activity, implicated in COVID-19 sequalae, was measured between two convalescent COVID-19 subjects, one with (Long-COVID subject) and another without Long-COVID symptoms. Methods: Flow adhesion of whole blood or isolated white blood cells to P-selectin (FA-WB-Psel and FA-WBC-Psel) was measured using a standardized microfluidics clinical assay; impedance aggregometry with a collagen agonist was measured using model 590 Chrono-Log impedance aggregometer; standard laboratory assays were performed to evaluate changes in blood chemistries. Results: For both subjects, hemoglobin, WBC, platelet counts, electrolytes and ferritin were within normal reference ranges, with FA-WB-Psel significantly elevated compared to healthy controls (p<0.01). In vitro treatment of whole blood samples with crizanlizumab (anti-p-selectin monoclonal antibody) within the clinical dose range (10 μg/ml) mL) inhibited FA-WB-Psel only in samples from asymptomatic post-COVID subject, with the Long-COVID subject sample requiring close to 5-fold elevated dose to achieve a response. Pronounced inhibition of P-selectin adhesion of isolated leukocytes was observed for both subjects in autologous platelet-poor plasma and buffer. Impedance aggregometry showed greater baseline platelet aggregation to collagen in the Long-COVID sample, although both samples responded similarly to aspirin-induced platelet inhibition. Conclusions: Presented results suggest that elevated platelet activation in Long-COVID subject may be associated with increased P-Selectin activity. The results are discussed in terms of possible use on P-selectin inhibition therapies in treating Long-COVID.

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Section: Methodsmentioning

confidence: 99%

Possible Role of P-selectin Adhesion in Long-COVID: A Comparative Analysis of a Long-COVID Case Versus an Asymptomatic Post-COVID Case

Tarasev

Gao

Mota

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

Background: Long-term outcomes of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are now recognized as an emerging public health challenge - a condition termed Long-COVID. The pathophysiology of Long-COVID remains to be established. Functional P-selectin activity, implicated in COVID-19 sequalae, was measured between two convalescent COVID-19 subjects, one with (Long-COVID subject) and another without Long-COVID symptoms. Methods: Flow adhesion of whole blood or isolated white blood cells to P-selectin (FA-WB-Psel and FA-WBC-Psel) was measured using a standardized microfluidics clinical assay; impedance aggregometry with a collagen agonist was measured using model 590 Chrono-Log impedance aggregometer; standard laboratory assays were performed to evaluate changes in blood chemistries. Results: For both subjects, hemoglobin, WBC, platelet counts, electrolytes and ferritin were within normal reference ranges, with FA-WB-Psel significantly elevated compared to healthy controls (p<0.01). In vitro treatment of whole blood samples with crizanlizumab (anti-p-selectin monoclonal antibody) within the clinical dose range (10 μg/ml) mL) inhibited FA-WB-Psel only in samples from asymptomatic post-COVID subject, with the Long-COVID subject sample requiring close to 5-fold elevated dose to achieve a response. Pronounced inhibition of P-selectin adhesion of isolated leukocytes was observed for both subjects in autologous platelet-poor plasma and buffer. Impedance aggregometry showed greater baseline platelet aggregation to collagen in the Long-COVID sample, although both samples responded similarly to aspirin-induced platelet inhibition. Conclusions: Presented results suggest that elevated platelet activation in Long-COVID subject may be associated with increased P-Selectin activity. The results are discussed in terms of possible use on P-selectin inhibition therapies in treating Long-COVID.

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“…Briefly, isolated white blood cell (i-WBC) suspensions were prepared according to a standardized protocol (HetaSep TM , StemCell Technologies). Whole blood samples (1:1 diluted with HBSS buffer) or i-WBC (5x 10 6 cells/mL) were perfused through P-selectin-coated microfluidic channels at 1 dynes/cm 2 for 10 or 6 minutes respectively, washed to eliminate non-adhering cells, with resultant adhesion quantified manually by an independent trained observer to generate an adhesion index (cells/mm²) using a previously described protocol [19]. For drug treatment conditions, samples were incubated for 5 minutes with crizanlizumab at the final concentrations as required before assessment by the flow adhesion assay.…”

Section: Study Subjects and Sample Collectionmentioning

confidence: 99%

Possible Role of P-selectin Adhesion in Long-COVID: A Case of Recovery After Long-COVID

Tarasev

Gao

Ferranti

et al. 2023

Preprint

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Background: Long-term outcomes of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are now recognized as an emerging public health challenge - a condition termed post-acute coronavirus 2019 syndrome (PACS) or Long-COVID. The pathophysiology of Long-COVID remains to be established, several mechanisms in study focus on the role of P-selectin, an inflammation-induced protein expressed by platelets and endothelial cells. Functional P-selectin activity, potentially implicated in COVID-19 and Long-COVID sequelae, was measured for a Long-COVID subject at 68 weeks from the SARS-CoV-2 infection after fully recover from the syndrome. It was compared with the results from the same subject at 20 weeks post-infection, when subject experienced severe Long-COVID symptoms. Methods: Flow adhesion of whole blood or isolated white blood cells to P-selectin (FA-WB-Psel and FA-WBC-Psel) was measured using a standardized microfluidics clinical assay; impedance aggregometry with a collagen agonist was measured using model 590 Chrono-Log impedance aggregometer; standard laboratory assays were performed to evaluate changes in blood chemistries. Results: After recovery from Long-COVID, RBC count and D-dimer remained elevated and other blood chemistry results remained within the normal range as compared to 20 weeks post infection when severe Long-COVID symptoms were present. Total iron and transferrin-iron saturation percentage values that were elevated when symptoms were present, declined to normal range. Whole blood aggregometry results indicate an absence of previously present platelet hyperactivity. FA-WB-PSel that was significantly elevated during Long-COVID (590 +/- 260 cells/mm2) was significantly reduced after the resolution of Long-COVID symptoms (98 +/- 38 cells/mm2). However, supplementation of whole blood with crizanlizumab did not result in any measurable inhibition of cell adhesion to P-selectin, similarly with previously reported. Similar to what was observed for the subject when Long-COVID symptoms were present, crizanlizumab, even at a dose 10-fold lower than clinical, induced pronounced inhibition of FA-WBC-Psel when tested in buffer, but not in patient own plasma. Conclusions: This report documents the changes in leukocyte adhesion properties for a patient at more than a year from the initial infection after the gradual resolution of Long-COVID symptoms, as compared to when Long-COVID symptoms were present. Recovery from Long-COVID may be associated with normalization of platelet activity, but not necessarily with complete alleviation of endothelial activation. It remains to be determined to what extent changes in leukocyte adhesion to P-selectin may represent a new risk factor for a mechanism driving Long-COVID syndrome.

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Assessment of antiplatelet therapy response in pediatric patients following cardiac surgery by microfluidic assay

Cited by 2 publications

References 39 publications

Possible Role of P-selectin Adhesion in Long-COVID: A Comparative Analysis of a Long-COVID Case Versus an Asymptomatic Post-COVID Case

Possible Role of P-selectin Adhesion in Long-COVID: A Comparative Analysis of a Long-COVID Case Versus an Asymptomatic Post-COVID Case

Possible Role of P-selectin Adhesion in Long-COVID: A Case of Recovery After Long-COVID

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