scholarly journals Activated Platelets Harbour SARS-CoV-2 During Severe COVID-19

Author(s):  
Ejaife O. Agbani ◽  
Prism Schneider ◽  
Braedon McDonald ◽  
Leslie Skeith ◽  
Man-Chiu Poon ◽  
...  

Clinical SARS-CoV-2 infection (COVID-19) is characterised by a hyperinflammatory and procoagulant state that increases the risk of thrombosis and death. Despite thromboprophylaxis at least at the conventional doses, incidence as high as 31% has been reported for thrombotic complications in Intensive Care Unit (ICU) patients with COVID-19 infections. Still, it remains unclear how SARS-Cov-2 may initiate and or perpetuates the intractable thrombo-inflammatory states in COVID-19; and the need to understand the underlying mechanisms is urgent. In this image report, we utilised a state-of-the-art high-resolution 3D imaging approach to examine the interactions of SARS-CoV-2 with platelets, erythrocytes and leucocytes in blood samples obtained from COVID-19 patients in our ICU; and we visualised platelet procoagulant activity and the spatial localisation of SARS-CoV-2 in platelet-rich-plasma reconstituted to contain erythrocytes and leucocytes. Strikingly, we observed that activated platelets harboured SARS-Cov-2 during severe COVID-19 in our patient that eventually succumbed to the infection. SARS-Cov-2 internalisation into the cytosol was probably via a passive mechanism, as we have previously established that actin cytoskeleton remodelling and increased membrane permeability occurred during platelet transformation to the procoagulant phenotype. More work is needed to understand platelets’ role in the recalcitrant thrombotic states of COVID-19.

1998 ◽  
Vol 79 (01) ◽  
pp. 177-185 ◽  
Author(s):  
Ashia Siddiqua ◽  
Michael Wilkinson ◽  
Vijay Kakkar ◽  
Yatin Patel ◽  
Salman Rahman ◽  
...  

SummaryWe report the characterization of a monoclonal antibody (MAb) PM6/13 which recognises glycoprotein IIIa (GPIIIa) on platelet membranes and in functional studies inhibits platelet aggregation induced by all agonists examined. In platelet-rich plasma, inhibition of aggregation induced by ADP or low concentrations of collagen was accompanied by inhibition of 5-hydroxytryptamine secretion. EC50 values were 10 and 9 [H9262]g/ml antibody against ADP and collagen induced responses respectively. In washed platelets treated with the cyclooxygenase inhibitor, indomethacin, PM6/13 inhibited platelet aggregation induced by thrombin (0.2 U/ml), collagen (10 [H9262]g/ml) and U46619 (3 [H9262]M) with EC50 = 4, 8 and 4 [H9262]g/ml respectively, without affecting [14C]5-hydroxytryptamine secretion or [3H]arachidonate release in appropriately labelled cells. Studies in Fura 2-labelled platelets revealed that elevation of intracellular calcium by ADP, thrombin or U46619 was unaffected by PM6/13 suggesting that the epitope recognised by the antibody did not influence Ca2+ regulation. In agreement with the results from the platelet aggregation studies, PM6/13 was found to potently inhibit binding of 125I-fibrinogen to ADP activated platelets. Binding of this ligand was also inhibited by two other MAbs tested, namely SZ-21 (also to GPIIIa) and PM6/248 (to the GPIIb-IIIa complex). However when tested against binding of 125I-fibronectin to thrombin stimulated platelets, PM6/13 was ineffective in contrast with SZ-21 and PM6/248, that were both potent inhibitors. This suggested that the epitopes recognised by PM6/13 and SZ-21 on GPIIIa were distinct. Studies employing proteolytic dissection of 125I-labelled GPIIIa by trypsin followed by immunoprecipitation with PM6/13 and analysis by SDS-PAGE, revealed the presence of four fragments at 70, 55, 30 and 28 kDa. PM6/13 did not recognize any protein bands on Western blots performed under reducing conditions. However Western blotting analysis with PM6/13 under non-reducing conditions revealed strong detection of the parent GP IIIa molecule, of trypsin treated samples revealed recognition of an 80 kDa fragment at 1 min, faint recognition of a 60 kDa fragment at 60 min and no recognition of any product at 18 h treatment. Under similar conditions, SZ-21 recognized fragments at 80, 75 and 55 kDa with the 55kDa species persisting even after 18 h trypsin treatment. These studies confirm the epitopes recognised by PM6/13 and SZ-21 to be distinct and that PM6/13 represents a useful tool to differentiate the characteristics of fibrinogen and fibronectin binding to the GPIIb-IIIa complex on activated platelets.


1984 ◽  
Vol 51 (01) ◽  
pp. 037-041 ◽  
Author(s):  
K M Weerasinghe ◽  
M F Scully ◽  
V V Kakkar

SummaryCollagen mediated platelet aggregation caused -5.6 ± 6.7% inhibition and +39.1 ± 15.2% potentiation of prekallikrein activation in plasma from normal healthy volunteers between 20–40 and 50–65 years of age, respectively (n = 15, p <0.01). The amouns of platelet factor-four (PF4) released in the two groups were not significantly different. Collagen treatment in the presence of indomethacin caused +11.5 ± 3.6% and +59.6 ± 19.5% potentiation in the 20–40 and 50–65 age groups respectively (p <0.02). Adrenaline mediated platelet aggregation caused -55.2 ± 7.1% and -35.2 ± 8.3% inhibition in the 20–40 and 50–65 age groups, respectively. Collagen treatment of platelet-deficient-plasma and platelet-rich-plasma in EDTA also caused potentiation of prekallikrein activation.The results indicate that the observed degree of prekallikrein activation after platelet aggregation is a net result of the inhibitory effect of PF4 and the potentiatory effect of activated platelets. The potentiatory effect was greater after collagen treatment as compared to adrenaline treatment, and in the 50–65 age group as compared to the 20–40 age group.


2021 ◽  
Vol 33 (6) ◽  
pp. 219-221
Author(s):  
Laura Bolton

Activated platelets release a rich broth of growth factors involved in wound healing. One way to deliver activated platelets to wounds is in the form of platelet-rich plasma (PRP) harvested by centrifuging the patient’s venous blood after activating the platelets with collagen or calcium chloride and/or autologous thrombin, then delicately removing the supernatant, called platelet-poor plasma (PPP). Platelet-rich plasma is usually injected into the lesion and/or applied topically, then sealed in or over the wound using a moisture-retentive dressing. Platelet-rich plasma (often with PPP) has been applied at different times, depths, and frequencies to chronic and acute wounds using various PRP doses and vehicles to achieve widely differing results. Meta-analyses have reported that PRP improved healing rates of open diabetic foot ulcers and venous ulcers1,2 and may reduce pain and surgical site infection (SSI) incidence in open and closed acute surgical wounds. However, inconsistency in study methods and outcome measures limited consistency of pain and SSI results.1 No consistent effect on healing or deep SSI rates was reported as a result of adding 1 intraoperative dose of PRP in the surgical site before closing elective foot and ankle surgery incisions of 250 patients as compared with 250 similar patients receiving the same procedure without PRP.3 After decades of research, ideal parameters of PRP delivery and use on each type of wound remain unclear for improving SSI, acute wound pain, and healing outcomes. This installment of the Evidence Corner reviews 2 surgical studies that may provide clues about optimal PRP use. One triple-blind randomized clinical trial (RCT) focused on irrigation of freshly closed carpal ligament surgical incisions with PRP as compared with PPP.4 Another non-blind RCT explored the effect of injecting PRP into open pilonidal sinus excisions 4 days and 12 days after surgery.5


Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 5-5
Author(s):  
Talat Almukhtar ◽  
Julio Hajdenberg ◽  
Gurjot Garcha

A test's negative predictive value-the probability that a negative result is a true negative result-is dependent on the prevalence of the condition. The present pandemic circumstances present us with unique challenges. False negatives in current testing methodology are to be expected.[1] Thus, a rigorous and contextual interpretation of a negative test result is necessary. Blood hypercoagulability and the risk of thrombosis are well documented in cases of the novel SARS-CoV-2 coronavirus (COVID-19) pandemic.[2] The systemic inflammatory response is associated with endothelial upregulation of proinflammatory mediators that lead to in situ thrombi, as well as a generalized disseminated intravascular coagulation. As many as 31% of ICU patients with COVID-19 have been reported to have thrombotic complications. More specifically, cerebral thrombotic complications confined to the arterial bed have been well described.[3, 4]. The case described shows that milder forms of coronavirus infection may lead to other types of critical and unusual thrombotic complications. An otherwise healthy 47-year-old Caucasian woman developed fever and respiratory signs and symptoms consistent with a possible case of COVID-19 infection in late March of 2020. Interstitial opacities were seen on radiographic examination. Two COVID-19 PCR nasopharyngeal tests were negative, and she recovered at home over the following 2 weeks. Three weeks later, she developed headaches, expressive aphasia, and a generalized tonic-clonic seizure. The patient was treated for a possible ischemic stroke with alteplase thrombolysis at a local hospital. After subsequent transfer and evaluation, a diagnosis of a left transverse and sigmoid sinus thrombosis with adjacent cortical edema was made. On review of her history, the patient denied taking any form of hormonal contraception, and did not have personal, or family history indicative of thrombophilia. She recovered fully after anticoagulation with enoxaparin and subsequent dabigatran. Prior to discharge a COVID-19, IgG antibody test was reported as positive. Decontextualized and overly simplistic interpretation of COVID-19 negative tests amidst a pandemic is problematic. In addition to the obvious infection control issues associated with the resulting lack of isolation and contract tracing, it may deprive some patients of the opportunity to receive antithrombotic therapy. Prophylactic and therapeutic regimens for hospitalized patients are in evolution, and have been associated with improved clinical outcomes.[5] We are aware that the role of anticoagulation in outpatient cases is not well studied, but we believe it deserves proper investigation. References: West, C.P., V.M. Montori, and P. Sampathkumar,COVID-19 Testing: The Threat of False Negative Results.Mayo Clin Proc, 2020.Thachil, J., et al.,ISTH interim guidance on recognition and management of coagulopathy inCOVID-19.J Thromb Haemost, 2020.18(5): p. 1023-1026.Oxley, T.J., et al.,Large-Vessel Stroke as a Presenting Feature of Covid-19 in the Young.N Engl J Med, 2020.Klok, F.A., et al.,Confirmation of the high cumulative incidence of thrombotic complications incritically ill ICU patients with COVID-19: An updated analysis.Thromb Res, 2020.Paranjpe, I., et al.,Association of Treatment Dose Anticoagulation with In-Hospital SurvivalAmong Hospitalized Patients with COVID-19.J Am Coll Cardiol, 2020. Disclosures No relevant conflicts of interest to declare.


1985 ◽  
Vol 249 (3) ◽  
pp. C279-C287 ◽  
Author(s):  
L. Salganicoff ◽  
M. H. Loughnane ◽  
R. W. Sevy ◽  
M. Russo

The ultrastructure and contractile behavior of a new preparation of thrombin-activated human platelets is described. The preparation is referred to as the "platelet strip" because of its similarities to classical vascular smooth muscle strips. The platelet strip consists of a giant platelet aggregate 10 mm long, 4 mm wide, and 200 micron thick. To facilitate handling, the aggregate has a special high-compliance nylon mesh embedded in its mass. Each strip contains 7.3 X 10(8) platelets. Fibrin contamination is 150-fold lower than in platelet-rich plasma clots. Active isometric forces of up to 100 g/cm2 and 6-10 h viability are easily and reproducibly obtained. Platelet strips remain contracted after thrombin activation. The contraction is tonic and partial. Further small increases in force can be produced by depolarizing solutions or pharmacological agents, e.g., ADP, epinephrine, and endoperoxide analogues. These small increases are reversible on washout of the agents. Full relaxation is induced by agents such as prostaglandin E1 or papaverine, which increase adenosine 3',5'-cyclic monophosphate. However, after washout of these agents, recovery of tension is variable depending on the concentration of the drug and the degree of prestretching of the preparation.


2020 ◽  
Vol 11 (3) ◽  
pp. 47
Author(s):  
Floris Honig ◽  
Steven Vermeulen ◽  
Amir A. Zadpoor ◽  
Jan de Boer ◽  
Lidy E. Fratila-Apachitei

The ability to control the interactions between functional biomaterials and biological systems is of great importance for tissue engineering and regenerative medicine. However, the underlying mechanisms defining the interplay between biomaterial properties and the human body are complex. Therefore, a key challenge is to design biomaterials that mimic the in vivo microenvironment. Over millions of years, nature has produced a wide variety of biological materials optimised for distinct functions, ranging from the extracellular matrix (ECM) for structural and biochemical support of cells to the holy lotus with special wettability for self-cleaning effects. Many of these systems found in biology possess unique surface properties recognised to regulate cell behaviour. Integration of such natural surface properties in biomaterials can bring about novel cell responses in vitro and provide greater insights into the processes occurring at the cell-biomaterial interface. Using natural surfaces as templates for bioinspired design can stimulate progress in the field of regenerative medicine, tissue engineering and biomaterials science. This literature review aims to combine the state-of-the-art knowledge in natural and nature-inspired surfaces, with an emphasis on material properties known to affect cell behaviour.


2020 ◽  
Vol 7 (1) ◽  
Author(s):  
Peng-Cheng Xu ◽  
Min Xuan ◽  
Biao Cheng

AbstractMilitary drill injury is a significant part of military medical research. The increase of training intensity and changes in training methods lead to differences in injury types. The ideal therapeutic modality should allow rapid healing at a reasonable cost and minimize impact on patients’ life. Platelet -rich plasma (PRP), a platelet concentrate, is rich in a variety of growth factors and widely used clinically as a minimally invasive treatment. It plays an important role in injury repair and rehabilitation. In this article, we review the therapeutic role of PRP in military drill injury and its possible underlying mechanisms, with a focus on plantar fasciitis, stress fractures and other common injuries, in order to provide basic support for military reserve.


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