Морфологическая и иммуногистохимическая характеристика аспирированных из сосудов тромбов у больных инфарктом миокарда с подъемом ST сегмента
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Sisakian H.S., Mkhitaryan A.G., Sahakyan V.G., et al. Morphological and Immunohystochemical Characteristics of Aspirated Thrombi in Patients with ST-Elevation Myocardial Infarction. Therapeutic Archive. 2019; 91 (9): 21–25. DOI: 10.26442/00403660.2019.09.000199
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Материалы и методы. Исследовали тромбы, полученные от 67 пациентов с инфарктом миокарда с подъемом ST сегмента. Проводили макроскопический и иммуногистохимический анализы для определения клеточных компонентов тромбов. Использовали иммуногистохимический метод пероксидаза-антипероксидазы с помощью моноклональных антител к CD4, CD8, CD15, CD63 и CD105 с использованием диаминобензидина в качестве визуализирующего агента для исследования потенциального вовлечения различных клеточных субпопуляций.
Результаты. Макроскопический анализ выявил рыхлые тромбы с видимыми слоями (lines of Zahn), которые при микроскопическом анализе выявлялись в виде рыхлой сетчатой структуры волокон фибрина с компактными скоплениями активированных тромбоцитов. Тромбоциты были окружены нейтрофильными гранулоцитами. Иммуногистохимический анализ подтвердил, что сгусток содержит CD63 позитивные активированные тромбоциты и лейкоциты. Гранулоциты были положительны к CD15-антигену. Иммуногистохимический анализ выявил также наличие CD8+ цитотоксических Т-лимфоцитов и CD4+ T-хелперных лимфоцитов. В анализированных тромбах не выявлено экспрессии iNOS, CD105 и сосудистого эндотелиального фактора роста.
Заключение. Гистопатологический анализ тромбов у пациентов с острым инфарктом миокарда с подъемом ST сегмента выявил возможное наличие активированных CD63+-тромбоцитов и CD15+-нейтрофильных гранулоцитов. Активированные тромбоциты и нейтрофилы могут играть важную патогенетическую роль в тромбовоспалительном процессе, приводящем к дестабилизации атеросклеротической бляшки и развитию острого тромбоза при остром инфаркте миокарда с подъемом ST сегмента.
Ключевые слова: инфаркт миокарда, коронарный тромбоз, активированные тромбоциты, тромбэктомия, иммуногистохимия.
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The aim of study was to evaluate the morphological and immunohystochemical characteristics of aspirated thrombi in patients with ST-elevation myocardial infarction undergoing precutaneous coronary intervention to find the possible platelet activation and inflammatory cell involvement in coronary thrombi.
Materials and methods. Thrombi collected from 67 patients with ST-elevation myocardial infarction were studied. Macroscopic and immunоhystochemical analyses were performed to reveal the cellular components of thrombi. The peroxidase-antiperoxidase immunohystochemical method was applied with monoclonal antibodies to CD4+, CD8+, CD15+, CD63+ and CD105+ using diaminobenzidine agent as a visualization agent to investigate the potential involvement of different cellular subpopulations.
Results. The macroscopic examination revealed friable thrombi with apparent laminations (lines of Zahn). Microscopic analysis of thrombi revealed a loose meshwork of fibrin fibers with compact accumulation of activated platelets. The platelets were surrounded by neutrophilic granulocytes.The immunohystochemical analysis confirmed that cell aggregates are consisting of CD 63+ activated platelets and leucocytes. The granulocytes also tested positive for CD15 antigen. The immunohystochemical analysis revealed the presence of CD8+ cytotoxic T-lymphocytes and CD4+ helper T-lymphocytes. No expression of iNOS, CD105 and VEGF was found in the analyzed thrombi.
Conclusions. The hystopathological evaluation of thrombi in patients with acute ST-elevation myocardial infarction revealed the presence of activated CD63+ platelets and CD15+ neutrophilic granulocytes. Activated platelets and neutrophils may play a role in thromboinflammatory activation course leading to destabilization of atherosclerotic plaque and development of acute thrombosis in patients with ST-elevation acute myocardial infarction.
Keywords: myocardial infarction, coronary thrombosis, activated platelets, thrombectomy, immunohistochemistry
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2. Srikanth S & Ambrose JA. Pathophysiology of coronary thrombus formation and adverse consequences of thrombus during PCI. Curr Cardiol Rev. 2012;8(3):168-76. doi: 10.2174/157340312803217247
3. Hansson GK, Libby P & Tabas I. Inflammation and plaque vulnerability. J Intern Med. 2015;278(5):483-93. doi: 10.1111/joim.12406
4. Ramaiola I, Padro T, Pena E, Juan-Babot O, Cubedo J, Martin-Yuste V, et al. Changes in thrombus composition and profilin-1 release in acute myocardial infarction. Eur Heart J. 2015;36(16):965-75. doi: 10.1093/eur heartj/ehu356
5. Sadowski M, Zabczyk M & Undas A. Coronary thrombus composition: links with inflammation, platelet and endothelial markers. Atherosclerosis. 2014;237(2):555-61. doi: 10.1016/j.atherosclerosis.2014.10.020
6. Silvain J, Collet JP, Nagaswami C, Beygui F, Edmondson KE, Bellemain-Appaix A, et al. Composition of coronary thrombus in acute myocardial infarction. J Am Coll Cardiol. 2011;57(12):1359-67. doi: 10.1016/j.jacc.2010.09.077
7. Yunoki K, Naruko T, Sugioka K, Inaba M, Iwasa Y, Komatsu R, et al. Erythrocyte-rich thrombus aspirated from patients with ST-elevation myocardial infarction: association with oxidative stress and its impact on myocardial reperfusion. Eur Heart J. 2012;33(12):1480-90. doi: 10.1093/ eurheartj/ehr486
8. [Kashtalap VV, Zavyrylina IN, Barbarash OL. Endovaskulyarnaya revaskulyarizatsiya pri ostrom koronarnom sindrome s pod"emom segmenta ST v Rossii: problemy i perspektivy dal'neishego razvitiya. Kreativnaya kardiologiya. 2015;3:5-15 (In Russ.)].
9. Windecker S, Kolh P, Alfonso F, Collet JP, Cremer J, et al. 2014 ESC/EACTS Guidelines on myocardial revascularization: The Task Force on Myocardial Revascularization of the European Society of Cardiology (ESC) and the European Association for Cardio-Thoracic Surgery (EACTS)Developed with the special contribution of the European Association of Percutaneous Cardiovascular Interventions (EAPCI). Eur Heart J. 2014;35(37):2541-619. doi: 10.1093/eurheartj/ehu278
10. Ibanez B, James S, Agewall S, Antunes MJ, Bucciarelli-Ducci C, Bueno H, et al. Group ESCSD. 2017 ESC Guidelines for the management of acute myocardial infarction in patients presenting with ST-segment elevation: The Task Force for the management of acute myocardial infarction in patients presenting with ST-segment elevation of the European Society of Cardiology (ESC). Eur Heart J. 2017. doi: 10.1093/eurheartj/ehx393
11. Cham BP, Gerrard JM & Bainton DF. Granulophysin is located in the membrane of azurophilic granules in human neutrophils and mobilizes to the plasma membrane following cell stimulation. Am J Pathol. 1994:144(6): 1369-80.
12. Kerr MA & Stocks SC. The role of CD15-(Le(X))-related carbohydrates in neutrophil adhesion. Histochem J. 1992;24(11):811-26.
13. Furman MI, Barnard MR, Krueger LA, Fox ML, Shilale EA, Lessard DM, et al. Circulating monocyte-platelet aggregates are an early marker of acute myocardial infarction. J Am Coll Cardiol. 2001;38(4):1002-6.
14. Michelson AD, Barnard MR, Krueger LA, Valeri CR, Furman MI. Circulating monocyte-platelet aggregates are a more sensitive marker of in vivo platelet activation than platelet surface P-selectin: studies in baboons, human coronary intervention, and human acute myocardial infarction. Circulation. 2001;104(13):1533-7.
15. [Trashchenko AS. Kliniko-immunologicheskie osobennosti techeniya infarkta miokarda s zubtsom Q: dis. … kand. med. nauk. Kemerovo, 2010: 122 s (In Russ.)].
16. Duerschmied D, Suidan GL, Demers M, Herr N, Carbo C, Brill A, et al. Platelet serotonin promotes the recruitment of neutrophils to sites of acute inflammation in mice. Blood. 2013;121(6):1008-15. doi: 10.1182/blood-2012-06-437392
17. Shi G, Field DJ, Long X, Mickelsen D, Ko KA, Ture S, et al. Platelet factor 4 mediates vascular smooth muscle cell injury responses. Blood. 2013;121(21):4417-27. doi: 10.1182/blood-2012-09-454710
18. Kramer MC, van der Wal AC, Koch KT, Rittersma SZ, Li X, Ploegmakers HP, et al. Histopathological features of aspirated thrombi after primary percutaneous coronary intervention in patients with ST-elevation myocardial infarction. PLoS One. 2009;4(6): e5817. doi: 10.1371/journal. pone.0005817
19. Rittersma SZ, van der Wal AC, Koch KT, Piek JJ, Henriques JP, Mulder KJ, et al. Plaque instability frequently occurs days or weeks before occlusive coronary thrombosis: a pathological thrombectomy study in primary percutaneous coronary intervention. Circulation. 2005;111(9):1160-5. doi: 10.1161/01.CIR.0000157141.00778.AC
20. Kovacs A, Sotonyi P, Nagy AI, Tenekedjiev K, Wohner N, Komorowicz E, et al. Ultrastructure and composition of thrombi in coronary and peripheral artery disease: correlations with clinical and laboratory findings. Thromb Res. 2015;135(4):760-6. doi: 10.1016/j.thromres.2015.02.004
21. Nishibori M, Cham B, McNicol A, Shalev A, Jain N & Gerrard JM. The protein CD63 is in platelet dense granules, is deficient in a patient with Hermansky-Pudlak syndrome, and appears identical to granulophysin. J Clinical Investigation. 1993;91(4):1775-82.
22. Israels SJ & McMillan-Ward EM. CD63 modulates spreading and tyrosine phosphorylation of platelets on immobilized fibrinogen. Thromb Haemost. 2005;93(2):311-8. doi: 10.1160/TH04-08-0503
23. Arakawa K, Yasuda S, Hao H, Kataoka Y, Morii I, Kasahara Y, et al. Significant Association Between Neutrophil Aggregation in Aspirated Thrombus and Myocardial Damage in Patients With ST-Segment Elevation Acute Myocardial Infarction. Circulation. 2009;73(1):139-44. doi: 10.1253/circj.CJ-08-0609
24. Ammirati E, Moroni F, Magnoni M & Camici PG. The role of T- and B-cells in human atherosclerosis and atherothrombosis. Clin Exp Immunol. 2015;179(2):173-87. doi: 10.1111/cei.12477
25. Sparks JA, Barbhaiya M, Karlson EW, Ritter SY, Raychaudhuri S, Corrigan CC, et al. Investigating methotrexate toxicity within a randomized double-blinded, placebo-controlled trial: Rationale and design of the Cardiovascular Inflammation Reduction Trial-Adverse Events (CIRT-AE) Study. Semin Arthritis Rheum. 2017;47(1):133-42. doi: 10.1016/j. semarthrit.2017.02.003
1 Ереванский государственный медицинский университет, Ереван, Республика Армения;
2 Клиника общей и инвазивной кардиологии университетской больницы №1 Ереванского государственного медицинского университета, Ереван, Республика Армения
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H.S. Sisakian 1,2, A.G. Mkhitaryan 1, V.G. Sahakyan 1, A.S. Kocharyan 1, B.A. Asatryan 1
1 Yerevan State Medical University, Yerevan, Armenia;
2 Clinic of General and Invasive Cardiology, University Hospital №1, Yerevan State Medical University, Yerevan, Armenia