KH.S. KHAERTYNOV¹, I.G. MUSTAPHIN¹, E.I. NASYROVA¹, A.T. TAGIEVA¹, G.F. MINGAZOVA²

 ¹Kazan State Medical University, Kazan

²Republic Clinical Hospital for Infectious Diseases named after Prof. A.F. Agafonov, Kazan

 Contact details:

Khaertynov Kh.S. — MD, Associate Professor of the Department of Children’s Infections

Address: 49 Butlerov St., 420012 Kazan, Russian Federation, tel.: +7-903-342-96-27, e-mail: khalit65@yandex.ru

 Seasonal influenza outbreaks are associated with increased incidence of myocardial infarction, stroke, and other cardiovascular complications. Endothelial dysfunction and platelet activation play significant roles in their pathogenesis. Platelet-leukocyte aggregates serve as promising markers of platelet activation, with their blood levels elevated in various cardiovascular diseases.

The purpose — to evaluate platelet activation in influenza patients by measuring platelet-neutrophil aggregates in peripheral blood.

Material and methods. Platelet activation was assessed in 18 influenza patients hospitalized at Republic Clinical Hospital for Infectious Diseases named after Prof. A.F. Agafonov between February 17, 2025 and April 7, 2025. The median patient age was 44 years. A control group included 10 age-matched healthy individuals. Platelet activation was determined by quantifying platelet-neutrophil aggregates in peripheral blood using immunofluorescence with fluorochrome-labeled monoclonal antibodies to CD antigens: CD61-FITC and CD13-PE (BD, USA).

Results. The acute phase of influenza in 78% of patients was associated with significantly increased formation of platelet-neutrophil aggregates compared to controls (р = 0.005). Thrombocytopenia (< 150 × 10⁹/L) was observed in 28% of cases. No correlation was found between platelet-neutrophil aggregate levels and platelet counts.

Conclusion. Influenza induces platelet activation, evidenced by an elevated number of platelet-neutrophil aggregates in blood, indicating increased risk of thrombotic complications.

Key words: influenza, platelets, leukocytes, granulocytes, platelet-neutrophil aggregates.

REFERENCES

1. Lemaitre M., Carrat F. Comparative age distribution of influenza morbidity and mortality during seasonal influenza epidemics and the 2009 H1N1 pandemic. BMC Infectious Diseases, 2010, vol. 10, p. 162. DOI: 10.1016/j.jcrc.2011.05.016
2. GBD 2017 Influenza Collaborators. Mortality, morbidity, and hospitalisations due to influenza lower respiratory tract infections, 2017: an analysis for the Global Burden of Disease Study 2017. Lancet Respir Med, 2019, vol. 7 (1), pp. 69–89. DOI: 10.1016/ S2213-2600(18)30496-X
3. Office WHOEMR. Global Influenza Strategy 2019–2030. Weekly Epidemiological Record, 2019.
4. Warren-Gash C., Blackburn R., Whitaker H. et al. Laboratory-confirmed respiratory infections as triggers for acute myocardial infarction and stroke: a self-controlled case series analysis of national linked datasets from Scotland. Eur. Respir. J, 2018, vol. 51 (3). 1701794. DOI: 10.1183/13993003.01794-2017
5. Ishmurzin G.P., Serebryakova O.A., Syuzev K.N. et al. Cardiovascular complications in respiratory viral infections. Sibirskiy zhurnal klinicheskoy i eksperimental’noy meditsiny, 2022, vol. 37, no. 4, pp. 31–37 (in Russ.). DOI: 10.29001/2073-8552-2022-37-4-31-37
6. Kwong J.C., Schwartz K.L., Campitelli M.A. et al. Acute Myocardial Infarction after Laboratory Confirmed Influenza Infection. N Engl. J. Med, 2018, vol. 378 (4), pp. 345–353. DOI: 10.1056/NEJMoa1702090
7. Chacko B., Peter J.V., Pichamuthu K. et al. Cardiac manifestations in patients with pandemic (H1N1) 2009 virus infection needing intensive care. J. Crit Care, 2012, vol. 27, p. 106. DOI: 10.1016/j.jcrc.2011.05.016
8. Rondina M.T., Brewster B., Grissom C.K., Zimmerman G.A. et al. In vivo platelet activation in critically ill patients with primary 2009 influenza A(H1N1). Chest, 2012, vol. 141, pp. 1490–1495. DOI: 10.1378/chest. 11-2860
9. Lisman T. Platelet–neutrophil interactions as drivers of inflammatory and thrombotic disease. Cell Tissue Res, 2018, vol. 371, pp. 567–576. DOI: 10.1007/s00441-017-2727-4
10. Michelson A.D., Barnard M.R., Krueger L.A. et al. 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, vol. 104, pp. 1533–1537. DOI: 10.1161/hc3801.095588
11. Yang D.H., Tan N., He P.C. et al. Increased platelet-leukocyte aggregates in patients with acute coronary syndrome. Zhonghua Xin Xue Guan Bing Za Zhi, 2012, vol. 40 (6), pp. 482–486.
12. Pluta K., Porebska K., Urbanowicz T. et al. Platelet–Leucocyte Aggregates as Novel Biomarkers in Cardiovascular Diseases. Biology, 2022, vol. 11, pp. 224. DOI: 10.3390/biology11020224
13. Armstrong S.M., Darwish I., Lee W.L. Endothelial activation and dysfunction in the pathogenesis of influenza A virus infection. Virulence, 2013, vol. 4 (6), pp. 537–542. DOI: 10.4161/viru.25779
14. Short K.R., Kuiken T., Van Riel D. Role of endothelial cells in the pathogenesis of influenza in humans. J. Infect. Dis, 2019, vol. 220 (11), pp. 1859–1860. DOI: 10.1093/infdis/jiz349
15. Marchenko V.A., Zhilinskaya I.N. Activation and dysfunction of the endothelium of blood vessels in infection caused by influenza A viruses (Alphainfluenzavirus influenzae). Voprosy virusologii, 2024, vol. 69, no. 6, pp. 465–478 (in Russ.). DOI: 10.36233/0507-4088-264
16. Filgueiras-Rama D., Vasilijevic J., Jalife J. et al. Human infuenza A virus causes myocardial and cardiac-specifc conduction system infections associated with early infammation and premature death. Cardiovasc Res, 2021, vol. 117 (3), pp. 876–889. DOI: 10.1093/cvr/cvaa117
17. Chang T.Y., Chao T.F., Liu C.J. et al. The association between influenza infection, vaccination, and atrial fibrillation: A nationwide case-control study. Heart Rhythm, 2016, vol. 13 (6), pp. 1189–1194. DOI: 10.1016/j.hrthm.2016.01.026
18. Ukimura A., Izumi T., Matsumori A. Clinical Research Committee on Myocarditis Associated with 2009 Influenza A (H1N1) Pandemic in Japan organized by Japanese Circulation Society. A national survey on myocarditis associated with the 2009 influenza A (H1N1) pandemic in Japan. Circ. J, 2010, vol. 74 (10), pp. 2193–2199. DOI: 10.1253/circj.cj-10-0452
19. Ergle K., GoodenJ.Y., Ahmed M.M. High-Grade atrioventricular block associated with acute influenza. Tex. Heart Inst. J, 2020, vol. 47 (3), pp. 220–223. DOI: 10.14503/THIJ-18-6658
20. Schrottmaier W.C., Schmuckenschlager A., Pirabe A. et al. Platelets in Viral Infections — Brave Soldiers or Trojan Horses. Front. Immunol, 2022, vol. 13. 856713. DOI: 10.3389/fimmu.2022.856713
21. Koupenova M., Corkrey H.A., Vitseva O. et al. The Role of Platelets in Mediating a Response to Human Influenza Infection. Nat. Commun, 2019, vol. 10 (1), p. 1780. DOI: 10.1038/s41467-019-09607-x
22. Jansen A.J.G., Spaan T., Low H.Z. et al. Influenza-induced thrombocytopenia is dependent on the subtype and sialoglycan receptor and increases with virus pathogenicity. Blood Adv, 2020, vol. 4 (13), pp. 2967–2978. DOI: 10.1182/bloodadvances.2020001640
23. Bahadoran A., Lee S.H., Wang S.M. et al. Immune Responses to Influenza Virus and Its Correlation to Age and Inherited Factors. Front. Microbiol, 2016, vol. 7, p. 1841. DOI: 10.3389/fmicb.2016.01841
24. Boilard E., Paré G., Rousseau M. et al. Influenza virus H1N1 activates platelets through FcγRIIA signaling and thrombin generation. Blood, 2014, vol. 123 (18), pp. 2854–2863. DOI: 10.1182/blood-2013-07-515536
25. Stegemann-Koniszewski S., Behrens S., Boehme J.D. et al. Respiratory influenza a virus infection triggers local and systemic natural killer cell activation via toll-like receptor 7. Front. Immunol, 2018, vol. 9, p. 245. DOI: 10.3389/fimmu.2018.00245
26. Blann A.D., Nadar S.K., Lip G.Y. The adhesion molecule P-selectin and cardiovascular disease. Eur. Hear. J, 2003, vol. 24, pp. 2166–2179. DOI: 10.1016/j.ehj.2003.08.021
27. Zimmerman G.A. Two by two: The pairings of P-selectin and P-selectin glycoprotein ligand 1. Proc. Natl. Acad. Sci. USA, 2001, vol. 98, pp. 10023–10024. DOI: 10.1073/pnas.191367898
28. Ivanov I.I., Apta B.H.R., Bonna A.M. et al. Platelet P-selectin triggers rapid surface exposure of tissue factor in monocytes. Sci. Rep, 2019, vol. 9 (1), p. 13397. DOI: 10.1038/s41598-019-49635-7
29. Gerrits A.J., Frelinger A.L. 3rd, Michelson A.D. Whole blood analysis of leukocyte-platelet aggregates. Curr. Protoc. Cytom, 2016, vol. 78, pp. 6.15.1–6.15.10. DOI: 10.1002/cpcy.8
30. Ferroni P., Martini F., Riondino S. et al. Soluble P-selectin as a marker of in vivo platelet activation. Clin. Chim. Acta, 2009, vol. 399 (1-2), pp. 88–91. DOI: 10.1016/j.cca.2008.09.018
31. Rolling C.C., Barrett T.J., Berger J.S. Platelet-monocyte aggregates: molecular mediators of thromboinflammation. Front. Cardiovasc. Med, 2023, vol. 10. 960398. DOI: 10.3389/fcvm.2023.960398
32. Barnes B.J., Adrover J.M., Baxter-Stoltzfus A. et al. Targeting potential drivers of COVID-19: Neutrophil extracellular traps. J. Exp. Med, 2020, vol. 217 (6). e20200652. DOI: 10.1084/jem.20200652
33. Middleton E.A., He X.-Y., Denorme F. et al. Neutrophil extracellular traps contribute to immunothrombosis in COVID-19 acute respiratory distress syndrome. Blood, 2020, vol. 136 (10), pp. 1169–1179. DOI: 10.1182/blood.2020007008
34. Ishiguro T., Matsuo K., Fujii S. et al. Acute thrombotic vascular events complicating influenza-associated pneumonia. Respir. Med. Case Rep, 2019, vol. 28. 100884. DOI: 10.1016/j.rmcr.2019.100884