Studying interconnections of cytokines and their diagnostic effectiveness when assessing severity grade in COVID-19 patients

The origins and intensity of human immune response to SARS-CoV-2 infection depend on many factors. The anti-infectious immune response includes both cellular and humoral components. The humoral link is mediated by effector-specific (antibodies) and non-specific factors (acute phase proteins, procalcitonin, α- and β-interferons), and regulatory factors, e.g., cytokines (interleukins (IL), γ-interferon (IFNγ)) and mediators (leukotrienes, etc.). In our work, serum levels of cytokines, i.e., IFNγ, IL-2, IL-6, IL-8, IL-10, IL-17A, IL-18 and procalcitonin (PCТ), a nonspecific effector factor of the immune system were compared at varying degrees of COVID-19 severity. Biomaterial samples (blood serum) of 91 patients aged 29-88 years (average age 53.9 years; 15 women, 76 males) were collected between May 2020 and July 2021. We divided patients into groups according to the disease severity, depending on clinical data, oxygen demand, and hospitalization data. COVID-19 patients of “mild degree” (n = 9) were subject to outpatient treatment; the patients of “moderate degree” (n = 38) were hospitalized at the infectious hospital; in the “severe degree” cases (n = 44), the patients were treated at the infectious intensive care unit. The control group of patients (n = 20) was presented by the blood donors without chronic diseases. We have identified the most important markers of the disease severity. Moreover, using ROC analysis, we have calculated the thresholds for differential diagnosis of distinct degrees of COVID-19 severity, by significantly differing criteria, as well as diagnostic efficiency of immune mediator indexes using a developed logistic regression model. We have revealed and statistically confirmed the indices that most significantly influence the COVID-19 severity, i.e., IL-6, IL-8, IL-10, IL-18, PCT, IFNγ. We found a maximum increase in IL-17A and IFNy at a mild degree, and their reduction in the moderate and maximum severe cases of COVID-19. Increase in IL-6, IL-10, PCT levels, and a decrease in IFNγ proved to be the factors of poor prognosis. We have also found some correlations between the immune mediators (IL-8, IL-17A, IL-18, IFNγ) thus allowing a deeper insight into the mechanisms of the disorders, the imbalance of immune response.

1. Bavrina A.P., Borisov I.B. Modern rules of the application of correlation analysis. Meditsinskiy almanakh = Medical Almanac, 2021, Vol. 68, no. 3, pp. 70-79. (In Russ.)

2. Bavrina A.P. Modern rules for the use of descriptive statistics methods in biomedical research. Meditsinskiy almanakh = Medical Almanac, 2020, Vol. 63, no. 2, pp. 95-105. (In Russ.)

3. Voeykova A.V., Rukavishnikova S.A., Zhukova N.L., Pushkin A.S., Akhmedov T.A., Saginbaev U.R., Volchkova E.V. Interleukin-6 and procalcitonin as prognostic clinical and laboratory parameters in COVID-19 in patients of olderage groups. Sovremennye problemy zdravookhraneniya i meditsinskoy statistiki = Current Problems of Health Care and Medical Statistics, 2022, no. 3, pp. 116-129. (In Russ.)

4. Grigoryeva N.Yu., Sinichkina A.A., Samoliuk M.O., Kolosova K.S., Korolеva E.V., Kondakova E.V., Vedunova M.V. Cytokine profile in hospitalized patients with COVID-19 of different severity. Rossiyskiy kardiologicheskiy zhurnal = Russian Jurnal of Cardiology, 2022, Vol. 27, no. 3, pp. 67-73. (In Russ.)

5. Zaytseva G.A.,Vershinina O.A., Matrokhina O.I., Senkina E.A., Karpova M.V. Cytokine status of the donors of blood and its components. Fundamentalnye issledovaniya = Fundamental Research, 2011, no. 3, pp. 61-65. (In Russ.)

6. Zelenskiy A.A., Efimenko N.A., Kazakov S.P. Features of changes in the level of cytokines and adhesion molecules in the development of early purulent-septic complications after surgery for gastric cancer. Infektsii v khirurgii = Infections in Surgery, 2008, Vol. 6, no. 1, pp. 40-44. (In Russ.)

7. Kazakov S.P. Differential levels of cytokines and intercellular adhesion molecules and their diagnostic efficiency in autoimmune and oncological thyroid diseases. Meditsinskay aimmunologiya = Medical Immunology (Russia), 2010, Vol. 12, no. 6, pp. 559-564. (In Russ.) doi: 10.15789/1563-0625-2010-6-559-564.

8. Kazakov S.P., Zabotina T.N., Kushlinskiy N.E. Ratios (indices) of cytokines, intercellular adhesion molecules in blood plasma, their diagnostic effectiveness in patients with autoimmune thyroiditis, adenomas and thyroid cancer. Meditsinskiy vestnik MVD = MIA Medical Bulletin, 2010, Vol. 49, no. 6, pp. 25-29. (In Russ.)

9. Kazakov S.P., Putkov S.B., Mudrov V.P., Zaitsev A.A., Troyan V.N. Systemic markers values of inflammation in assessing the severity of COVID-19 patients. Laboratornaya meditsina = Laboratory Medicine, 2022, no. 13, pp. 7-14. (In Russ.)

10. Kovalyova A.Ya., Nikolaev K.Yu., Agamian E.R., Lifshits G.I. Role of procalcitonin in diagnosis and prognosis of pneumonia caused by the new coronavirus infection 2019 (COVID-19). Sibirskoye meditsinskoye obozreniye = Siberian Medical Review, 2023, Vol. 143, no. 5, pp. 74-80. (In Russ.)

11. Kozlov V.A., Savchenko A.A., Kudryavtsev I.V., Kozlov I.G., Kudlay D.A., Prodeus A.P., Borisov A.G. Clinical immunology: a practical guide for doctors. Krasnoyarsk: Polikor, 2020, 386 p.

12. Kudryashov S.K., Kanishchev Yu.N., Putkov S.B., Esaulenko N.B., Karpov V.O., Ovcharenko V.P., Izgorodin A.S., Zhukova E.E., Suslova L.A., Parshakova E.V. Instructions for conducting the preanalytical stage (procedure for taking, storing and transporting) with biomaterial for laboratory research at the Center for Clinical Laboratory Diagnostics of the Main Military N. Burdenko Clinical Hospital, Russian Defense Ministry / Ed. Kazakov S.P. Moscow: Eko-Press, 2016, Vol. 51. 220 p.

13. Lutckii A.A., Zhirkov A.A., Lobzin D.Yu., Rao M., Alekseeva L.A., Maeurer M., Lobzin Yu.V. Interferon-γ: biological function and application for study of cellular immune response. Zhurnal infektologii = Jurnal of Infectology, 2015, Vol. 7, no. 4, pp. 10-22. (In Russ.)

14. Putkov S.B., Davydova N.V., Kazakov S.P. Study of inflammatory markers and their diag-nostic effectiveness in patients with COVID-19 patients. Meditsinskiy alfavit = Medical Alphabet, 2023, no. 23, pp. 11-17. (In Russ.)

15. Putkov S.B., Davydova N.V., Troyan V.N., Kazakov S.P. Investigation of the relationship of inflammatory markers with CT signs of pathological lung changes in patients with COVID-19. Meditsinskiy vestnik GVKG im. N.N. Burdenko = Medical Bulletin of the Main Military N. Burdenko Clinical Hospital, 2024, Vol. 5, no. 2, pp. 22-31. (In Russ.)

16. Sanitary and epidemiological rules SP 3.1.3597-20 “Prevention of new coronavirus infection (COVID-19)”. [Electronic resource]. Available at: https://docs.cntd.ru/document/564979137?ysclid = lzid31zq6w18848903.

17. Sakhin V.T., Kryukov E.V., Grigoriev M.A., Kazakov S.P., Sotnikov A.V., Gordienko A.V., Gulyaev N.I., Rukavitsyn O.A. Features of the secretion of interleukin-6, interleukin-1β, interferon-γ and their influence on the development of anemia of chronic diseases in patients with rheumatic pathology. Meditsinskiy vestnik GVKG im. N.N. Burdenko = Medical Bulletin of the Main Military N. Burdenko Clinical Hospital, 2021, no. 2 (4), pp. 26-33. (In Russ.)

18. Sakhin V.T., Kryukov E.V., Kazakov S.P., Sotnikov A.V., Gordienko A.V., Zavartsev M.A., Rukavitsyn O.A. Comparison of cytokine secretion and study of their effect on erythropoiesis in patients with malignant neoplasms with anemia of chronic disease, iron deficiency anemia, and their combination. Onkogematologiya = Oncohematology, 2023, Vol. 18, no. 2, pp. 45-52. (In Russ.)

19. Sakhin V.T., Kryukov E.V., Grigoryev M.A., Kazakov S.P., Sotnikov A.V., Gordienko A.V., Rukavitsyn O.A. Iron metabolism, cytokine secretion in patients with rheumatologic pathology. Klinicheskaya meditsina = Clinical Medicine, 2020, Vol. 98, no. 9-10, pp. 691-698. (In Russ.)

20. Tuguz A.R., Vershinina M.V. The ratio of the main cytokines in the blood of cancer patients and healthy donors. Vestnik Adygeyskogo gosudarstvennogo universiteta = Bulletin of Adyghe State University, 2007, no. 4. [Electronic resource]. Available at: http://vestnik.adygnet.ru/files/2008.1/632/tuguz2008_1.pdf. (In Russ.)

21. Khaitov R.M., Skvortsova V.I., Khaitov M.R. Biomedical safety: immunogenetics and coronavirus infection. Moscow: GEOTAR-Media, 2022. 352 p.

22. Yashkina O.N., Tsareva N.G., Borisova O.V. Correlations of interleukin levels with the severity of SARS-CoV-2 coronavirus infection. Detskie infektsii = Children Infections, 2023, Vol. 22, no. 1, pp. 11-13. (In Russ.)

23. Basheer M., Saad E., Kananeh M., Asad L., Khayat O., Badarne A., Abdo Z., Arraf N., Milhem F., Bassal T., Boulos M., Assy N. Cytokine patterns in COVID-19 patients: which cytokines predict mortality and which protect against? Curr. Issues Mol. Biol., 2022, Vol. 44, no. 10, pp. 4735-4747.

24. Cesta M.C., Zippoli M., Marsiglia C., Gavioli E.M., Mantelli F., Allegretti M., Balk R.A. The role of interleukin-8 in lung inflammation and injury: implications for the management of COVID-19 and hyperinflammatory acute respiratory distress syndrome. Front. Pharmacol., 2022, Vol. 12, 808797. doi: 10.3389/fphar.2021.808797.

25. Frey N., Porter D. Cytokine release syndrome with chimeric antigen receptor T cell therapy. Biol. Blood Marrow Transplant., 2019, Vol. 25, no. 4, pp. e123-e127.

26. Ghanbari Naeini L., Abbasi L., Karimi F., Kokabian P., Abdi Abyaneh F., Naderi D. The important role of interleukin-2 in COVID-19. J. Immunol. Res., 2023, Vol. 2023, 7097329. doi: 10.1155/2023/7097329.

27. Han H., Ma Q., Li C., Liu R., Zhao L., Wang W., Zhang P., Liu X., Gao G., Liu F., Jiang Y., Cheng X., Zhu C., Xia Y. Profiling serum cytokines in COVID-19 patients reveals IL-6 and IL-10 are disease severity predictors. Emerg. Microbes Infect., 2020, Vol. 9, no. 1, pp. 1123-1130.

28. Han J.M., Patterson S.J., Levings M.K. The role of the PI3K signaling pathway in CD4(+) T cell differentiation and function. Front. Immunol.,2012, Vol. 3, 245. doi: 10.3389/fimmu.2012.00245.

29. Hawerkamp H.C., Dyer A.H., Patil N.D., McElheron M., O’Dowd N., O’Doherty L., Mhaonaigh A.U., George A.M., O’Halloran A.M., Reddy C., Kenny R.A., Little M.A., Martin-Loeches I., Bergin C., Kennelly S.P., Donnelly S.C., Bourke N.M., Long A., Sui J., Doherty D.G., Conlon N., Cheallaigh C.N., Fallon P.G. Characterisation of the pro-inflammatory cytokine signature in severe COVID-19. Front. Immunol., 2023, Vol. 14, 1170012. doi: 10.3389/fimmu.2023.1170012.

30. Luo X.H., Zhu Y., Mao J., Du R.C. T cell immunobiology and cytokine storm of COVID-19. Scand. J. Immunol., 2021, Vol. 93, no. 3, e12989. doi: 10.1111/sji.12989.

31. Ng T.H., Britton G.J., Hill E.V., Verhagen J., Burton B.R., Wraith D.C. Regulation of adaptive immunity; the role of interleukin-10. Front. Immunol., 2013, Vol. 4, 129. doi: 10.3389./fimmu.2013.00129.

32. Queiroz M.A.F., Neves P.F.M.D., Lima S.S., Lopes J.D.C., Torres M.K.D.S., Vallinoto I.M.V.C., Bichara C.D.A., dos Santos E.F., de Brito M.T.F.M., da Silva A.L.S., Leite M.M., da Costa F.P., Viana M.N.D.S.A., Rodrigues F.B.B., de Sarges K.M.L., Cantanhede M.H.D., da Silva R., Bichara C.N.C., van den Berg A.V.S., Veríssimo A.O.L., Carvalho M.D.S., Henriques D.F., Dos Santos C.P., Nunes J.A.L., Costa I.B., Viana G.M.R., Carneiro F.R.O., Palacios V.R.D.C.M., Quaresma J.A.S., Brasil-Costa I., Dos Santos E.J.M., Falcão L.F.M., Vallinoto A.C.R. Cytokine Profiles Associated With Acute COVID-19 and Long COVID-19 Syndrome. Front. Cell. Infect. Microbiol., 2022, Vol. 12, 922422. doi: 10.3389/fcimb.2022.922422.

33. Satış H., Özger H.S., Aysert Yıldız P., Hızel K., Gulbahar Ö., Erbaş G., Aygencel G., Guzel Tunccan O., Öztürk M.A., Dizbay M., Tufan A. Prognostic value of interleukin-18 and its association with other inflammatory markers and disease severity in COVID-19. Cytokine, 2021, Vol. 137, 155302. doi: 10.1016/j.cyto.2020.155302.

34. Sorrentino L., Fracella M., Frasca F., D’Auria A., Santinelli L., Maddaloni L., Bugani G., Bitossi C., Gentile M., Ceccarelli G., Turriziani O., Mastroianni C.M., Antonelli G., d’Ettorre G., Pierangeli A., Scagnolari C. Alterations in the expression of IFN lambda, IFN gamma and toll-like receptors in severe COVID-19 patients. Microorganisms, 2023, Vol. 11, no. 3, 689. doi: 10.3390/microorganisms11030689.

35. Tiwari V., Agarwal J., PathakA.K., Singh S. Dynamic changes in circulatory cytokines and chemokines levels in mild to severe COVID-19 patients. Indian J. Clin. Biochem., 2023, Vol. 38, no. 2, pp. 212-219.

36. WHO. COVID-19 Epidemiological Update – 27 October 2023. Available at: https://www.who.int/pubLications/m/item/covid-19-epidemioLogicaL-update---27-october-2023.

37. Zanza C., Romenskaya T., Manetti A.C., Franceschi F., La Russa R., Bertozzi G., Maiese A., Savioli G., Volonnino G., Longhitano Y. Cytokine Storm in COVID-19: Immunopathogenesis and Therapy. Medicina (Kaunas), 2022, Vol. 58, no. 2, 144. doi: 10.3390/medicina58020144.