Platelet Reactivity to Zika and Dengue Non-Structural Protein 1 (NS1) Assessed by Flow Cytometry, Atomic Force Microscopy, and Quartz Crystal Microbalance

IF 2.2 4区医学 Q3 HEMATOLOGY International Journal of Laboratory Hematology Pub Date : 2024-12-05 DOI:10.1111/ijlh.14409

Alan Cano-Méndez, Gabriel Espinosa, Nallely García-Larragoiti, Pedro Antonio Maciel-García, Jorge Luis Menchaca-Arredondo, Young Chan-Kim, Arturo Reyes-Sandoval, Martha Eva Viveros-Sandoval

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Abstract

Background

Platelets, besides being traditionally associated with hemostasis, have been recently positioned as immune cells. Alterations in platelet number and function have been reported in some viral infections. Zika virus (ZIKV) and Dengue virus (DENV) are arboviruses that encode for a non-structural protein 1 (NS1). NS1 is mainly involved in the viral replication process and can also be secreted by infected cells and has been associated with immune response evasion. The assessment of platelet reactivity against these viral agents and their proteins, through the use of different innovative technologies such as flow cytometry (FC), atomic force microscopy (AFM), and quartz crystal microbalance (QCM), will allow further study of the pathophysiology of these emerging diseases.

Aim

The aim of this study was to assess platelet reactivity to ZIKV and DENV NS1 protein through the use of FC, AFM, and QCM.

Methods

Platelet-rich plasma (PRP) was stimulated with ZIKV and DENV NS1 protein in individual assays. The expression of P-selectin and the activity of the glycoprotein IIb-IIIa, platelet activation markers, were assessed by FC, morphological changes were assessed by AFM, and interaction between NS1 protein and platelet were evaluated by QCM.

Results

An increased expression of P-selectin and GP IIb-IIIa activity (p < 0.001) was observed when PRP was stimulated with ZIKV and DENV NS1 proteins. AFM images showed an increase in cell size and the appearance of pseudopods upon stimulation with the viral proteins. QCM results showed a significant increase in the oscillation frequency of the quartz precoated with ZIKV or DENV NS1 when PRP was injected (p < 0.001).

Conclusion

FC, AFM, and QCM are techniques that can be used in the study of platelet response to viral structures such as NS1 protein, broadening the range of existing methodologies in the study of these cells. It is imperative to study platelets in arboviral infections to better understand their involvement in these diseases.

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用流式细胞术、原子力显微镜和石英晶体微天平评估血小板对寨卡和登革热非结构蛋白1 （NS1）的反应性

背景：血小板，除了传统上与止血有关外，最近被定位为免疫细胞。一些病毒感染中有血小板数量和功能改变的报道。寨卡病毒（ZIKV）和登革热病毒（DENV）是编码非结构蛋白1 （NS1）的虫媒病毒。NS1主要参与病毒复制过程，也可由感染细胞分泌，与免疫应答逃避有关。通过使用不同的创新技术，如流式细胞术（FC）、原子力显微镜（AFM）和石英晶体微天平（QCM），评估血小板对这些病毒制剂及其蛋白质的反应性，将有助于进一步研究这些新发疾病的病理生理学。目的：本研究的目的是通过FC、AFM和QCM评估血小板对ZIKV和DENV NS1蛋白的反应性。方法：分别用ZIKV和DENV NS1蛋白刺激富血小板血浆（PRP）。FC检测p -选择素的表达及血小板活化标志物糖蛋白IIb-IIIa的活性，AFM检测形态学变化，QCM检测NS1蛋白与血小板的相互作用。结论：FC、AFM和QCM技术可用于研究血小板对NS1蛋白等病毒结构的反应，拓宽了研究这些细胞的现有方法的范围。研究虫媒病毒感染中的血小板以更好地了解它们在这些疾病中的作用是必要的。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

International Journal of Laboratory Hematology 医学-血液学

CiteScore

4.50

自引率

6.70%

发文量

211

审稿时长

6-12 weeks

期刊介绍： The International Journal of Laboratory Hematology provides a forum for the communication of new developments, research topics and the practice of laboratory haematology. The journal publishes invited reviews, full length original articles, and correspondence. The International Journal of Laboratory Hematology is the official journal of the International Society for Laboratory Hematology, which addresses the following sub-disciplines: cellular analysis, flow cytometry, haemostasis and thrombosis, molecular diagnostics, haematology informatics, haemoglobinopathies, point of care testing, standards and guidelines. The journal was launched in 2006 as the successor to Clinical and Laboratory Hematology, which was first published in 1979. An active and positive editorial policy ensures that work of a high scientific standard is reported, in order to bridge the gap between practical and academic aspects of laboratory haematology.