Features of modification of the proinflammatory cytokine profile by chemical and biological factors in an in vitro experiment (using benz(a)pyrene and influenza virus antigens as an example)

Introduction. Investigating formation of specific immune phenotypes associated with cytokine profile features in viral infections upon their additional modification by exposure to chemical factors will provide better insight into pathways of immune dysfunction associated with environmental factors.

Materials and methods. A mixed population of immunocytes was isolated and cultivated for 48 hours, and the cytokine profile was analyzed by enzyme immunoassay. Inducing factors were represented by benzo(a)pyrene and viral vaccine antigens hemagglutinins of influenza viruses type A and B.

Results. The multidirectional effects of benzo(a)pyrene on the production of pro-inflammatory cytokine mediators were established under exposure "in vitro" conditions with activation of TNF-α and MCP-1 expression and inhibition of IL-8 production (p=0.001–0.018). Modification of cytokine expression in the presence of influenza viral antigens led to preferential stimulation of cytokine production (IL-1β, IL-18, IL-8, IFN-γ, TNF-α, MCP-1) (p=0.015–0.045). Simultaneous introduction of benzo(a)pyrene and influenza virus antigens into the cell culture was accompanied by declining expression of mediators IL-1β, IL-8 and IFN-γ (p=0.005–0.047). Thus, influenza virus antigens stimulate production of pro-inflammatory mediators, while experimental benzo(a)pyrene burden adversly modifies biological effects of viral antigens, inhibiting cytokine expression.

Limitations. The limitations of the methodological approach are related to the need to further verify the obtained results and to extrapolate them to the macroorganism level.

Conclusion. The study findings show the high modifying activity of benzo(a)pyrene affecting the pro-inflammatory cytokine profile, canceling the additional expression of IL-1β, IL-8 and IFN-γ mediators induced by influenza antigens and reducing the protective immune potential. The revealed features of the combined effects produced by benzo(a)pyrene and influenza virus antigens, verified in an in vitro experiment, will allow specifying the mechanisms and peculiarities of the impact of chemical and biological factors to solve the problems of prevention of viral diseases under additional threats and risks associated with chemical exposures (exemplified by benzo(a)pyrene).

Compliance with ethical standards. All donors signed voluntary informed consent to participate in the study. The study was carried out in accordance with the Helsinki Declaration of the World Medical Association (revised 2013) and the Convention for the Protection of Human Rights and Dignity of the Human Being with regard to the Application of Biology and Medicine: Convention on Human Rights and Biomedicine (1999), according to the principles of “Good Clinical Practice” (GOST R 52379–2005) and approved by the Local Ethics Committee of the Federal Scientific Center for Medical and Preventive Technologies for Public Health Risk Management (Protocol No. 3 dated March 20, 2024).

Contribution:
Zaitseva N.V. – study concept and design, editing the text;
Starkova K.G. – study concept and design of the, data collection and analysis, writing the text;
Dolgikh O.V. – study concept and design, editing the text.
All authors are responsible for the integrity of all parts of the manuscript and approval of the manuscript final version.

Conflict of interest. The authors declare no conflict of interest.

Funding. The study had no sponsorship.

Received: April 24, 2026 / Accepted: June 18, 2026 / Published: July 31, 2026

1. WHO. Exposure & health impacts of air pollution. Available at: https://who.int/teams/environment-climate-change-and-health/air-quality-energy-and-health/health-impacts/exposure-air-pollution

2. Chang D., Dela Cruz C., Sharma L. Beneficial and detrimental effects of cytokines during influenza and COVID-19. Viruses. 2024; 16(2): 308. https://doi.org/10.3390/v16020308 https://elibrary.ru/aqvpum

3. Kazantseva E.D., Petrova A.G., Rychkova L.V., Darenskaya M.A. Cytokines in influenza in children (literature review). Sibirskii nauchnyi meditsinskii zhurnal. 2024; 44(1): 124–38. https://doi.org/10.18699/SSMJ20240113 https://elibrary.ru/exdfkd (in Russian)

4. Wei F., Gao C., Wang Y. The role of influenza A virus-induced hypercytokinemia. Crit. Rev. Microbiol. 2022; 48(2): 240–56. https://doi.org/10.1080/1040841X.2021.1960482 https://elibrary.ru/cialrd

5. Jaakkola J.J.K. Invited perspective: on air pollution, epidemics, pandemics, and planetary health. Environ. Health Perspect. 2023; 131(12): 121306. https://doi.org/10.1289/EHP13800 https://elibrary.ru/odzsqv

6. Loaiza-Ceballos M.C., Marin-Palma D., Zapata W., Hernandez J.C. Viral respiratory infections and air pollutants. Air Qual. Atmos. Health. 2022; 15(1): 105–14. https://doi.org/10.1007/s11869-021-01088-6 https://elibrary.ru/bhowwm

7. Provenzano S., Roth S., Sager L. Air pollution and respiratory infectious diseases. Environ. Resource Econ. 2024; 87(5): 1127–39. https://doi.org/10.1007/s10640-024-00858-x https://elibrary.ru/qycqdp

8. Xu B., Lim J.T., Chen C. Association between ambient air pollutants and major infectious diseases in Singapore. Environ. Chall. 2025; 21: 101297. https://doi.org/10.1016/j.envc.2025.101297 https://elibrary.ru/tvebai

9. Bukowska B., Mokra K., Michałowicz J. Benzo[a]pyrene – environmental occurrence, human exposure, and mechanisms of toxicity. Int. J. Mol. Sci. 2022; 23(11): 6348. https://doi.org/10.3390/ijms23116348 https://elibrary.ru/xzpyat

10. Bukowska B., Duchnowicz P. Molecular mechanisms of action of selected substances involved in the reduction of benzo[a]pyrene-induced oxidative stress. Molecules. 2022; 27(4): 1379. https://doi.org/10.3390/molecules27041379 https://elibrary.ru/japjom

11. Zhou B., Wang L., Yang S., Liang Y., Zhang Y., Pan X., et al. Diosmetin alleviates benzo[a]pyrene-exacerbated H1N1 influenza virus-induced acute lung injury and dysregulation of inflammation through modulation of the PPAR-γ-NF-κB/P38 MAPK signaling axis. Food Funct. 2023; 14(7): 3357–78. https://doi.org/10.1039/d2fo02590f https://elibrary.ru/uxpwzk

12. Gu Y., Zuo X., Zhang S., Ouyang Z., Jiang S., Wang F., et al. The mechanism behind influenza virus cytokine storm. Viruses. 2021; 13(7): 1362. https://doi.org/10.3390/v13071362 https://elibrary.ru/ivkywz

13. Zaitseva N.V., Zemlyanova M.A., Koldibekova Yu.V. Cellular-molecular and systemic biomarkers in hygiene [Kletochno-molekulyarnye i sistemnye biomarkery v gigiene]. Perm; 2025. 744 p. (in Russian)

14. Márquez-Bandala A.H., Gutierrez-Xicotencatl L., Esquivel-Guadarrama F. Pathogenesis induced by influenza virus infection: role of the early events of the infection and the innate immune response. Viruses. 2025; 17(5): 694. https://doi.org/10.3390/v17050694 https://elibrary.ru/arrnpu

15. Betakova T., Kostrabova A., Lachova V., Turianova L. Cytokines induced during influenza virus infection. Curr. Pharm. Des. 2017; 23(18): 2616–22. https://doi.org/10.2174/1381612823666170316123736 https://elibrary.ru/yejrfr

16. Karaba A.H., Zhou W., Hsieh L.L., Figueroa A., Massaccesi G., Rothman R.E., et al. Differential cytokine signatures of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) and Influenza infection highlight key differences in pathobiology. Clin. Infect. Dis. 2022; 74(2): 254–62. https://doi.org/10.1093/cid/ciab376 https://elibrary.ru/eeqrsw

17. Monoson A., Schott E., Ard K., Kilburg-Basnyat B., Tighe R.M., Pannu S., et. al. Air pollution and respiratory infections: the past, present, and future. Toxicol. Sci. 2023; 192(1): 3–14. https://doi.org/10.1093/toxsci/kfad003 https://elibrary.ru/yoqtmm

18. Poniedziałek B., Rzymski P., Zarębska-Michaluk D., Flisiak R. Viral respiratory infections and air pollution: a review focused on research in Poland. Chemosphere. 2024; 359: 142256. https://doi.org/10.1016/j.chemosphere.2024.142256 https://elibrary.ru/ttgofi

19. Zaitseva N.V., Starkova K.G., Dolgikh O.V., Shirinkina A.S. Features of cytokine expression under its modification with vaccine viral antigens (SARS-CoV-2 and Influenzavirus) and benzo(a)pyrene in experimental models in vitro. Gigiena i Sanitaria (Hygiene and Sanitation, Russian journal). 2025; 104(3): 353–7. https://doi.org/10.47470/0016-9900-2025-104-3-353-357 https://elibrary.ru/mcohej (in Russian)

20. Coumoul X., Barouki R., Esser C., Haarmann-Stemmann T., Lawrence B.P., Lehmann J., et al. The aryl hydrocarbon receptor: structure, signaling, physiology and pathology. Signal Transduct. Target. Ther. 2026; 11(1): 20. https://doi.org/10.1038/s41392-025-02500-8 https://elibrary.ru/zbfahm

21. Fueldner C., Kohlschmidt J., Riemschneider S., Schulze F., Zoldan K., Esser C., et. al. Benzo(a)pyrene attenuates the pattern-recognition-receptor induced proinflammatory phenotype of murine macrophages by inducing IL-10 expression in an aryl hydrocarbon receptor-dependent manner. Toxicology. 2018; 409: 80–90. https://doi.org/10.1016/j.tox.2018.07.011 https://elibrary.ru/vgiwyz

22. Fueldner C., Riemschneider S., Haupt J., Jungnickel H., Schulze F., Zoldan K., et al. Aryl hydrocarbon receptor activation by benzo[a]pyrene prevents development of septic shock and fatal outcome in a mouse model of systemic Salmonella enterica infection. Cells. 2022; 11(4): 737. https://doi.org/10.3390/cells11040737 https://elibrary.ru/yipfub

23. Schmidt J.R., Haupt J., Riemschneider S., Kämpf C., Löffler D., Blumert C., et al. Transcriptomic signatures reveal a shift towards an anti-inflammatory gene expression profile but also the induction of type I and type II interferon signaling networks through aryl hydrocarbon receptor activation in murine macrophages. Front. Immunol. 2023; 14: 1156493. https://doi.org/10.3389/fimmu.2023.1156493 https://elibrary.ru/ksflzc

24. Onishchenko G.G., Zaitseva N.V., eds. Health Risk Analysis in the Strategy of State Socio-Economic Development [Analiz riska zdorov’yu v strategii gosudarstvennogo social’no-ehkonomicheskogo razvitiya]. Moscow-Perm’; 2024. (in Russian)