Induction and mechanisms of antitumor activity of human macrophages upon combined activation of NOD- and Toll-like receptors
AbstractIntroduction. The treatment of malignant tumors is one of the most urgent tasks of modern medicine. Reprogramming of tumor-associated macrophages into antitumor effector cells is a promising direction in immunotherapy of cancer. Strong inducers of the antitumor activity of macrophages are signals from the pattern-recognition receptors (PRR) of innate immunity. Previously, we showed that the combination of NOD1 and TLR4 agonists effectively induces macrophage activity against erythromyeloid leukemia cells K562 [1].
The aim of the work was to study activity of human macrophages stimulated by a NOD1 agonist in combination with a TLR4 or a TLR7/8 agonist against cell lines derived from disseminated and solid tumors, as well as to analyze the mechanisms of antitumor activity of reprogrammed macrophages.
Material and methods. Erythromyeloid K562 cells, human epithelioid cervical carcinoma HeLa cells and human colorectal carcinoma HT29 cells carrying green fluorescent protein (GFP) gene were used as targets for macrophages. Tumor cells were coincubated with macrophages in the presence of NOD1, TLR4, TLR7/8 agonists separately and in combination. Tumor cells without macrophages were used as controls. To analyze the antiproliferative properties of macrophages, the cell cycle of tumor cells was analyzed using flow cytometry. The number of dead cells was determined by staining with annexin-V and propidium iodide. TNF mRNA expression in macrophages was evaluated by real-time reverse transcript PCR.
Results. Combinations of NOD1+TLR4 and NOD1+TLR7/8 agonists effectively induced macrophage activity against K562, HeLa and HT29 cells. The activity of agonist combinations in all cases was higher than the activity of individual agonists. Macrophages activated by a combination of NOD1+TLR4 agonists inhibited the proliferation of K562 tumor cells and enhanced their death.
Discussion contains an analysis of the data obtained and an assessment of the applicability of the proposed models of reprogramming macrophages in relation to various cell lines.
Conclusion. Combinations of NOD- and Toll-like receptor agonists effectively induce macrophage activity against tumor cell lines derived from disseminated and solid tumors.
Keywords: macrophages; antitumor immunity; K562 cells; HeLa cells; HT29 cells; pattern-recognition receptors; NOD1; TLR4; TLR7/8; tumor necrosis factor; apoptosis; proliferation; cell cycle
For citation: Esipova D.D., Murugin V.V, Poletkina A.A, Pashenkov M.V, Murugina N.E. Induction and mechanisms of antitumor activity of human macrophages with combined activation of NOD- and Toll-like receptors. Immunologiya. 2024; 45 (4): 486–94. DOI: https://doi.org/10.33029/1816-2134-2024-45-4-486-494 (in Russian)
Funding. The study was supported by the grant of Russian Science Foundation No. 21-15-00211.
Conflict of interests. The authors declare no conflict of interests.
Authors’ contribution. Concept and design of the study – Murugina N.E., Pashchenkov M.V.; collection and processing of material – Esipova D.D., Murugin V.V., Poletkina A.A.; statistical processing – Esipova D.D.; writing the text – Esipova D.D., Murugina N.E.; editing – Pashchenkov M.V.
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