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

  1. Michael Tarasev
  2. Sabrina Mota
  3. Xiufeng Gao
  4. Marta Ferranti
  5. Aliya U. Zaidi
  6. Bryan Hannan
  7. Patrick Hines

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Abstract

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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  1. ScreenIT

    SciScore for 10.1101/2022.03.09.22271297: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    EthicsIRB: Study Subjects and Sample Collection: Two convalescent COVID-19 subjects, one with Long-COVID symptoms (LC) and the other asymptomatic post-COVID (PC) were recruited according to the protocols FF-RBC-001 and FF-RBC-003v2 approved by Institutional Review Board of the Institute for Regenerative and Cellular Medicine.
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Whole blood was collected into EDTA tube for CBC, aggregometry, and flow adhesion assays, 3.2% sodium citrate tube for D-dimer, and serum separation tubes for interleukins, COVID-19 antibodies and for calcium, LDH, iron, TIBC and ferritin panel that were transferred to Quest Diagnostic Lab for analysis.
    COVID-19
    suggested: None
    LDH
    suggested: None

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:
    The limitation of this discussion is that the possibility of leukocyte-platelet aggregates co-purifying during i-WBC isolation cannot be completely ruled out. It remains uncertain whether SARS-CoV-2 is capable of established a persistent and stable chronic infection in human tissues like other SARS coronaviruses [54]. Nevertheless, viral RNA is known to persist in human body well past the resolution of the active infection [3] and SARS-CoV-2 RNA had been identified in samples from patients long time after the resolution of the active infection [55-57]. Considering that SARS-CoV-2 RNA present in the blood stream was associated with platelet hyperactivity [58, 59], a similar mechanism for platelet activation can be suggested for Long-COVID case. The key limitation of such a hypothesis would derive from the fact that the above studies observed SARS-CoV-2 RNA up to a maximum of 3 months after the infection, while in the presented case, the Long-COVID patient (LC) presented platelet hyperactivity at 20 weeks and experienced severe Long-COVID symptoms even at 28 weeks after the resolution of active COVID-19 disease. In that regard, the potential for persistent SARS-CoV-2 or presence of active viral RNA in patient body more than half a year post resolution had not been fully assessed. The uncertain potential for disease resurgence and/or spread gives this question extra importance and urgency. The clear limitation of this work is the observation of a single Long-COVID patient, even ...

    Results from TrialIdentifier: We found the following clinical trial numbers in your paper:

    IdentifierStatusTitle
    NCT04435184CompletedCrizanlizumab for Treating COVID-19 Vasculopathy

    Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

    Results from JetFighter: We did not find any issues relating to colormaps.

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • No protocol registration statement was detected.

    Results from scite Reference Check: We found no unreliable references.

    About SciScore

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  2. Version published to 10.1101/2022.03.09.22271297 on medRxiv