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
{"title":"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","authors":"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","doi":"10.1111/ijlh.14409","DOIUrl":null,"url":null,"abstract":"<div>\n \n \n <section>\n \n <h3> Background</h3>\n \n <p>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.</p>\n </section>\n \n <section>\n \n <h3> Aim</h3>\n \n <p>The aim of this study was to assess platelet reactivity to ZIKV and DENV NS1 protein through the use of FC, AFM, and QCM.</p>\n </section>\n \n <section>\n \n <h3> Methods</h3>\n \n <p>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.</p>\n </section>\n \n <section>\n \n <h3> Results</h3>\n \n <p>An increased expression of P-selectin and GP IIb-IIIa activity (<i>p</i> < 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 (<i>p</i> < 0.001).</p>\n </section>\n \n <section>\n \n <h3> Conclusion</h3>\n \n <p>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.</p>\n </section>\n </div>","PeriodicalId":14120,"journal":{"name":"International Journal of Laboratory Hematology","volume":"47 2","pages":"246-254"},"PeriodicalIF":2.2000,"publicationDate":"2024-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Platelet Reactivity to Zika and Dengue Non-Structural Protein 1 (NS1) Assessed by Flow Cytometry, Atomic Force Microscopy, and Quartz Crystal Microbalance\",\"authors\":\"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\",\"doi\":\"10.1111/ijlh.14409\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div>\\n \\n \\n <section>\\n \\n <h3> Background</h3>\\n \\n <p>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.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Aim</h3>\\n \\n <p>The aim of this study was to assess platelet reactivity to ZIKV and DENV NS1 protein through the use of FC, AFM, and QCM.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Methods</h3>\\n \\n <p>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.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Results</h3>\\n \\n <p>An increased expression of P-selectin and GP IIb-IIIa activity (<i>p</i> < 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 (<i>p</i> < 0.001).</p>\\n </section>\\n \\n <section>\\n \\n <h3> Conclusion</h3>\\n \\n <p>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.</p>\\n </section>\\n </div>\",\"PeriodicalId\":14120,\"journal\":{\"name\":\"International Journal of Laboratory Hematology\",\"volume\":\"47 2\",\"pages\":\"246-254\"},\"PeriodicalIF\":2.2000,\"publicationDate\":\"2024-12-05\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"International Journal of Laboratory Hematology\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1111/ijlh.14409\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"HEMATOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"International Journal of Laboratory Hematology","FirstCategoryId":"3","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1111/ijlh.14409","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"HEMATOLOGY","Score":null,"Total":0}
Platelet Reactivity to Zika and Dengue Non-Structural Protein 1 (NS1) Assessed by Flow Cytometry, Atomic Force Microscopy, and Quartz Crystal Microbalance
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.
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.
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