The effect of cfDNA on the expression of TLR receptors in human mesenchymal stem cells

Background. TLR-mediated activation of the innate immune response varies with cell type. TLRs can recognize not only exogenous pathogenic molecules (PAMP - pathogen-associated molecular patterns), but also endogenous molecules that appear during tissue damage, aseptic inflammation, and degeneration - DAMP. Under certain circumstances, this reaction may be uncontrollable, which leads to the development of severe systemic inflammation and sepsis. TLR9 is the only TLR that is capable of detecting pathogenic CpG DNA in endolysosomal structures. GC-rich rDNA fragments, which are TLR9 ligands, are accumulated in eDNA during pathology, pregnancy, or under the action of damaging factors. Aim: to investigate the influence of cfDNA fragments of different composition on the expression of TLR9 and on the expression of other human TLRs in in vitro cultures. Methods. The study was carried out on histologically different cultures with different proliferative potentials: mesenchymal stem cells (N = 13), HUVEC (N = 7), and MCF7 human breast adenocarcinoma cells. The expression of surface proteins by cells was studied by flow cytometry. To simulate the effect of eDNA on different types of cells, there were prepared model forms of DNA: genomic DNA (hydrolyzed by nuclease), oxidized forms of gDNA, and a model GC-enriched DNA fragment - CpG - a rich fragment of the transcribed region of rDNA. The expression level of TLR 1-10 genes was assessed by real-time PCR. Results. We noticed an increase in the expression of intracellular endoplasmic receptors TLR3, TLR7 and TLR8 genes in response to the action of both GC-rich and oxidized cfDNA fragments in different types of cells. In addition, the presence of cfDNA increases the expression of cell surface receptors TLR6 genes, and, to a lesser extent, TLR1 and TLR5. Under the action of oxidized fragments, the expression of the TLR4 gene increases. However, an increase in the expression of TLR family genes occurs secondarily after TLR9 activation. Blocking TLR9 inhibits signal transduction through other TLRs. Conclusions. Binding of An increase in the expression of receptors of the TLR family, except for TLR9, observed upon exposure to cfDNA, may be associated with the involvement of the TLR network in the regulation of signal transduction through TLR9, both through interactions between TLR receptors and through other signaling pathways associated with cfDNA recognition by DNA sensors.

cell free DNA, TLR

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