Lipid rafts and their role in altering monocyte properties in the blood of women who had COVID-19 during the third trimester of pregnancy

Introduction. Current literature widely addresses issues related to the pathogenesis of COVID-19 during pregnancy. However, the problem of dysfunction in the monocyte/macrophage system in pregnant women, particularly concerning the influence of changes in the lipid membrane microenvironment caused by SARS-CoV-2, remains unresolved. Aim. To conduct a comparative study and explore the association of lipid rafts with the expression of CD receptors on monocytes involved in forming the immune response in women who had COVID-19 during pregnancy. Materials and methods. The study included women with mild (n = 25) and moderate (n = 27) severity of COVID-19 in the third trimester of pregnancy, and 25 women not infected with SARS-CoV-2 during pregnancy. Using flow cytometry, lipid rafts on blood monocytes were identified by the intensity of the cholera toxin B-subunit (CTB)/ganglioside GM1 complex formation, as well as the expression of Fcγ receptor II (CD32), mannose receptor (CD206), tumor necrosis factor receptors type 1 (TNFR1) and type 2 (TNFR2), interleukin 17 receptor (IL17R), and TNF-related apoptosis-inducing ligand (TRAIL). Lipid raft microscopy was performed using a fluorescent microscope. Results. An increase in the distribution density and number of rafts in the monocyte membrane was established, which were 1.6 times higher (p < 0.001) in moderate disease severity compared to mild cases. The expression levels of CD206 increased by 1.8 times (p < 0.001), CD32 by 1.05 times (p < 0.05), TNFR1 by 1.2 times (p < 0.001), IL17R by 1.7 times (p < 0.001), and TRAIL by 1.4 times (p < 0.001) compared to mild disease. No differences in TNFR2 expression were found between subgroups (p = 0.781). A direct correlation was identified between lipid raft expression levels and CD206 (ρ = 0.70, p < 0.01), CD32 (ρ = 0.77, p < 0.01), TNFR1 (ρ = 0.63, p < 0.01), IL17R (ρ = 0.60, p < 0.01), and TRAIL (ρ = 0.70, p < 0.01). An inverse correlation was also established between the gestational age at delivery and the expression of rafts (ρ = -0.53, p < 0.01), CD206 (ρ = -0.36, p = 0.008), and CD32 (ρ = -0.32, p = 0.02). However, the gestational age at the time of illness was not associated with changes in the expression of lipid rafts and CD receptors. Conclusion. In women who had COVID-19 during the third trimester of pregnancy, monocytes predominantly exhibit a pro-inflammatory phenotype expressing increased amounts of pre-activation markers CD206 and CD32, as well as cytokine receptors TNFR1, IL17R, and TRAIL. It can be hypothesized that the increased expression of CD206, CD32, and IL17R—which directly correlated with the number of lipid rafts—may be directly related to monocyte activation and, thus, to the severity of the infection and the development of complications during pregnancy.

1. Adamyan L.V., Vechorko V.I., Filippov O.S., Konysheva O.V., Kharchenko E.I., Fattahova D.N. [Novel coronavirus infection (COVID-19). Labor outcomes for women with and without COVID-19 during a pandemic (data of the obstetric department of the Filatov City Clinical Hospital No. 15)]. Problemy Reproduktsii = Russian Journal of Human Reproduction 2021; 27(3-2):15–22 (in Russian). https://doi.org/10.17116/repro20212703215

2. Zhukovets I.V., Аndrievskaya I.A., Кrivoshchekova N.A., Smirnova N.A., Petrova K.K., Kharchenko M.V., Nikachalo D.A. [First effects of the COVID-19 pandemic: pregnancy complications, newborn health and expected reproductive losses]. Bûlleten' fiziologii i patologii dyhaniâ = Bulletin of Physiology and Pathology of Respiration 2022; 84:77-85 (in Russian). https://doi.org/10.36604/1998-5029-2022-84-77-85

3. Voropaeva E.E., Khaidukova Y.V., Kazachkova E.A., Kazachkov E.L., Shamaeva T.N., Aliyeva A.A., Ishchenko L.S., Holopova A.Y., Sychugov G.V. [Perinatal outcomes and morphological examination of placentas in pregnant women with critical lung lesions in new COVID-19 coronavirus infection]. Ural'skij medicinskij zhurnal = Ural Medical Journal 2023; 22(2):109-121 (in Russian). https://doi.org/10.52420/2071-5943-2023-22-2-109-121

4. Malinowski A. K., Noureldin A., Othman M. COVID-19 susceptibility in pregnancy: Immune/inflammatory considerations, the role of placental ACE-2 and research considerations. Reprod. Biol. 2020; 20(4):568–572. https://doi.org/10.1016/j.repbio.2020.10.005

5. Menter T., Mertz K.D., Jiang S., Chen H., Monod C., Tzankov A., Waldvogel S., Schulzke S.M., Hösli I., Bruder E. Placental pathology findings during and after SARSCoV-2 Infection: features of villitis and malperfusion. Pathobiology 2021; 88(1):69–77. https://doi.org/10.1159/000511324

6. Redline R.W., Ravishankar S., Bagby C., Saab S., Zarei S. Diffuse and localized SARS-CoV-2 placentitis: prevalence and pathogenesis of an uncommon complication of COVID-19 Infection during pregnancy. Am. J. Surg. Pathol. 2022; 46(8):1036–1047. https://doi.org/10.1097/PAS.0000000000001889

7. Andrievskaya I.A., Lyazgyan K.S. [Expression of CD68 by macrophages and histopathology of the placenta in COVID-19: association with obstetric and neonatal complications]. Bûlleten' fiziologii i patologii dyhaniâ = Bulletin of Physiology and Pathology of Respiration 2024; 93:91–99 (in Russian). https://doi.org/10.36604/1998-5029-2024-93-91-99

8. Martínez-Diz S., Marín-Benesiu F., López-Torres G., Santiago O., Díaz-Cuéllar J.F., Martín-Esteban S., Cortés-Valverde A.I., Arenas-Rodríguez V., Cuenca-López S., Porras-Quesada P., Ruiz-Ruiz C., Abadía-Molina A.C., Entrala-Bernal C., Martínez-González L.J., Álvarez-Cubero M.J. Relevance of TMPRSS2, CD163/CD206, and CD33 in clinical severity stratification of COVID-19. Front. Immunol. 2023; 13:1094644. https://doi.org/10.3389/fimmu.2022.1094644

9. Knoll R., Schultze J.L., Schulte-Schrepping J. Monocytes and Macrophages in COVID-19. Front. Immunol. 2021; 12:720109. https://doi.org/10.3389/fimmu.2021.720109

10. Radyukhin V.A., Baratova L.A. Molecular mechanisms of raft organization in biological membranes. Russian Journal of Bioorganic Chemistry 2020; 46(3):269–279. https://doi.org/10.31857/S0132342320030264

11. Vitner E.B., Avraham R., Politi B., Melamed S., Israely T. Elevation in sphingolipid upon SARS-CoV-2 infection: possible implications for COVID-19 pathology // Life Sci. Alliance. 2021. Vol.5, Iss.1. Article number:e202101168. https://doi.org/10.26508/lsa.202101168

12. Shen W., Stone K., Jales A., Leitenberg D., Ladisch S. Inhibition of TLR activation and up-regulation of IL-1Rassociated kinase-M expression by exogenous gangliosides // J. Immunol. 2008. Vol.180, Iss.7. P.4425–4432. https://doi.org/10.4049/jimmunol.180.7.4425

13. Kim S.J., Chung T.W., Choi H.J., Jin U.H., Ha K.T., Lee Y.C., Kim C.H. Monosialic ganglioside GM3 specifically suppresses the monocyte adhesion to endothelial cells for inflammation // Int. J. Biochem. Cell Biol. 2014. Vol.46. P.32– 38. https://doi.org/10.1016/j.biocel.2013.09.015

14. Sonnino S., Mauri L., Chigorno V., Prinetti A. Gangliosides as components of lipid membrane domains // Glycobiology. 2007. Vol.17, Iss.1. P.1R–13R. https://doi.org/10.1093/glycob/cwl052.

15. Lingwood D., Simons K. Lipid rafts as a membrane-organizing principle // Science. 2010. Vol.327(5961). P.46– 50. https://doi.org/10.1126/science.1174621.

16. Radyukhin V.A., Dadinova L.A., Orlov I.A., Baratova L.A. Amphipathic secondary structure elements and putative cholesterol recognizing amino acid consensus (CRAC) motifs as governing factors of highly specific matrix protein interactions with raft-type membranes in enveloped viruses // J. Biomol. Struct. Dyn. 2018. Vol.36. P.1351–1359. https://doi.org/10.1080/07391102.2017.1323012

17. Schmitz G., Orsó E. CD14 signalling in lipid rafts: new ligands and co-receptors // Curr. Opin. Lipidol. 2002. Vol.13, Iss.5. P.513–521. https://doi.org/10.1097/00041433-200210000-00007

18. Barnett K.C., Kagan J.C. Lipids that directly regulate innate immune signal transduction // Innate Immun. 2020. Vol.26, Iss.1. P.4–14. https://doi.org/10.1177/1753425919852695

19. Triantafilou M., Miyake K., Golenbock D.T., Triantafilou K. Mediators of innate immune recognition of bacteria concentrate in lipid rafts and facilitate lipopolysaccharide-induced cell activation // J. Cell Sci. 2002. Vol.115, Iss.12. P.2603–2611. https://doi.org/10.1242/jcs.115.12.2603

20. Gizzi A.S., Grove T.L., Arnold J.J., Jose J., Jangra R.K., Garforth S.J., Du Q., Cahill S.M., Dulyaninova N.G., Love J.D., Chandran K., Bresnick A.R., Cameron C.E., Almo S.C. A naturally occurring antiviral ribonucleotide encoded by the human genome // Nature. 2018. Vol.558, Iss.7711. P.610–614. https://doi.org/10.1038/s41586-018-0238-4

21. Gabrilovich D., Nagaraj S. Myeloid-derived suppressor cells as regulators of the immune system // Nat. Rev. Immunol. 2009. Vol.9, Iss.3. P.162–174. https://doi.org/10.1038/nri2506

22. Arsentieva N.A., Batsunov O.K., Kudryavtsev I.V., Semenov A.V., Totolian A.A. [CD32a receptor in health and disease]. Meditsinskaya Immunologiya = Medical Immunology (Russia) 2020; 22(3):433-442 (in Russian). https://doi.org/10.15789/1563-0625-CRI-2029

23. Andrievskaya I.A., Lyazgiyan K.S., Zhukovets I.V., Ustinov E.M. [Effect of COVID-19 infection in the third trimester of pregnancy on innate immunity parameters, association with obstetric and perinatal outcomes]. Bûlletenʹ sibirskoj mediciny = Bulletin of Siberian Medicine 2024; 23(2):5-13 (in Russian). https://doi.org/10.20538/1682-0363-2024-2-5-13

24. Jin Z., El-Deiry W.S. Distinct signaling pathways in TRAIL-versus tumor necrosis factor-induced apoptosis // Mol. Cell. Biol. 2006. Vol.26, Iss.21. P.8136–8148. https://doi.org/10.1128/MCB.00257-06

25. Di Pietro R., Zauli G. Emerging non-apoptotic functions of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)/Apo2L // J. Cell. Physiol. 2004. Vol.201, Iss.3. P.331–340. https://doi.org/10.1002/jcp.20099

26. Zingler P., Särchen V., Glatter T., Caning L., Saggau C., Kathayat R.S., Dickinson B.C., Adam D., Schneider-Brachert W., Schütze S., Fritsch J. Palmitoylation is required for TNF-R1 signaling // Cell Commun. Signal. 2019. Vol.17, Iss.1. Article number:90. https://doi.org/10.1186/s12964-019-0405-8

27. Ponde N.O., Shoger K.E., Khatun M.S., Sarkar M.K., Dey I., Taylor T.C., Cisney R.N., Arunkumar S.P., Gudjonsson J.E., Kolls J.K., Gottschalk R.A., Gaffen S.L. SARS-CoV-2 ORF8 mediates signals in macrophages and monocytes through MyD88 independently of the IL-17 receptor // J. Immunol. 2023. Vol.211, Iss.2. P.252–260. https://doi.org/10.4049/jimmunol.2300110