Synergism of TLR3 and TLR4 agonists during macrophage reprogramming into an antitumor stateстатья
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Статья опубликована в журнале из списка Web of Science и/или Scopus
Дата последнего поиска статьи во внешних источниках: 8 апреля 2022 г.
Аннотация:AbstractIntroduction. Solid tumor tissue usually contains many macrophages. These cells actively support the progressive growth of malignant cells, stimulate angiogenesis, promote the growth of the fi brotic scaffold the tumor by activating divisions of fi broblasts and their transformationinto myofi broblasts, immune responses inhibition with immunosuppressive substances such as IL-10, TGFβ, IDO, PGE2, and an induction of epithelial-mesenchymal transformationof malignant cells, which allows malignant cells to move freely within tissues (invasive tumor growth) and around the host organism (distant metastases). Macrophages’ plasticity, i.e. their ability for transition from one state to another, is well documented by previous studies. Particularly,tumor-promoting macrophages can be reprogrammed into anti-tumor M1 macrophages. External infection-derived signals acting via pathogen recognition receptors (TLR, NOD, etc.), as well as internal damage-derived signals acting via pro-infl ammatory cytokine and other receptors,transfer macrophages into M1 state.The aim of this study is to analyze the synergism of TLR3- and TLR4-agonists during macrophages reprogramming into their M1 antitumor state.Material and methods. Macrophages were obtained from the mouse bone marrow using in vitro differentiation in the presence of GM-CSF. Cells were activated with synthetic poly I:C (TLR3 agonist) or acidic peptidoglycan of the plant origin (TLR4 agonist, active componentof Immunomax, hereinafter IMM). The activation of TLR3- and TLR4-related downstream signaling pathways was measured according to TBK1/IKKε, ERK1 and ERK2 protein kinases, as well as NF-κB transcription factor phosphorylation intensities. Flow cytometry and Westernblotting with specifi c monoclonal antibodies were used to measure phosphorylation degree of the signaling proteins. The expression intensity of Nos2 and Ifnb1 mRNAs was investigated by a quantitative RT-PCR. The cytotoxic activity of macrophages was estimated in their co-culturewith 4T1 breast carcinoma cells according to the reduction of the tumor cells number.Results. A transformation of macrophages into an antitumor state can be induced with a combination of TLR3- and TLR4-agonists, more effi ciently than with the same agonists used alone. Both agonists induced signifi cant and rapid activation of TBK1/IKKε, ERK1/2 and NF-κB. Compared to the single agonists, the combined action of TLR3 and TLR4 agonistsincreased the kinases phosphorylation level by only 15–20 %. Even though the activated genes transcription rate was enhanced synergistically. The combined action of TLR3 and TLR4 agonistsincreased the level of Nos2 and Ifnb1 mRNAs transcription by 2.6–3.5 times as compared to the transcription rate of the same genes in response to each of these agonists used alone.TLR3- and TLR4-agonists also synergistically activated the antitumor properties of macrophages in a cytotoxic test against 4T1 carcinoma cells.