The amniotic variant of glycodelin (Gd) has pronounced immunomodulatory properties and is involved in the formation of immune tolerance during pregnancy. The role of recombinant Gd at physiological (0.2 and 2 μg/ml) and superphysiological (10 μg/ml) concentrations in regulating the differentiation and functional activity of human myeloid-derived suppressor cells (MDSCs) was investigated in vitro. MDSCs were generated from CD11b+ peripheral blood cells of healthy donors by two-step induction (IL-1β + GM-CSF and then lipopolysaccharide (LPS). The effect of Gd on the content of polymorphonuclear MDSC (PMN-MDSC) and monocytic MDSC (M-MDSC), intracellular expression of indoleamine 2.3-dioxygenase (IDO), arginase-1 (Arg1, and cytokine profile in cell cultures was investigated. In general, the transformation of CD11b+ cells into MDSCs exhibits the following characteristics: as a result of cytokine induction, predominantly M-MDSCs but no PMN-MDSCs are formed and Arg1 expression is virtually undetected. Gd was found to increase the number of M-MDSCs at concentrations of 2 and 10 μg/ml. Gd was found not to affect Arg1 expression but increased the percentage of MDSCs expressing IDO (10 μg/ml). Gd also modulated the cytokine profile of CD11b+ cells by suppressing the production of IL-19, IL-26 and TWEAK/TNFsF12 at a physiological concentration of 2 μg/ml and the production of IFN-α2 and IL-26 at a supraphysiological concentration. Thus, the role of Gd in the conversion of CD11b+ cells to MDSCs was examined under conditions of cytokine induction in vitro.
Amitriptyline is a tricyclic antidepressant widely used in clinical practice for the treatment of anxiety, depression and chronic pain. These drugs have a multifaceted effect on cellular processes. One of their targets is sigma-1 receptors. Sigma-1 receptors are molecular chaperones located in endoplasmic reticulum membrane; they are characterized by a unique structure and pharmacological profile. Sigma-1 receptors regulate many cellular processes in health and disease, including Ca2+ signaling. Using Fura-2AM microfluorimetry, it was shown for the first time that sigma-1 receptor agonist, antidepressant amitriptyline, significantly suppresses both Ca2+ mobilization from intracellular Ca2+-stores and subsequent store-dependent Ca2+ entry into cells, induced by endoplasmic Ca2+-ATPase inhibitors thapsigargin and cyclopiazonic acid, as well as disulfide-containing immunomodulators glutoxim and molixan, in rat peritoneal macrophages. The results suggest the involvement of sigma-1 receptors in a complex signaling cascade induced by glutoxim or molixan, leading to an increase of intracellular Ca2+ concentration in macrophages. The results also indicate the participation of sigma-1 receptors in the regulation of store-dependent Ca2+ entry in macrophages.
Voltage-gated cardiac sodium channels Nav1.5 are responsible for the initiation and propagation of action potentials in cardiomyocytes. Dysfunction of Nav1.5 can be caused both by pathogenic variants in the SCN5A gene itself, which encodes Nav1.5, and by genetic variants in the genes of other proteins, regulating channel activity and trafficking. The change of different phases of the action potential is determined by the strict temporal organization of activation and inactivation of various ion channels. Transitions between channel functional states (for example, to slow inactivated state) can be influenced by various factors and proteins interacting with the channel. Despite the fact that the process of slow inactivation of the channel has been known for several decades, its role in the mechanism of development of hereditary heart pathology remains unclear. In this work, using the patch clamp method in whole-cell leads, we studied changes in the process of slow Nav1.5 inactivation under the influence of various mutations in structural genes (DSP-H1684R, LMNA-R249Q, FLNC-R1267Q, FLNC-V2264M) associated with a genetically determined myocardial pathology leading to dysfunction of cardiomyocytes. The study used a model of cardiomyocytes differentiated from induced pluripotent stem cells (СM-iPSCs). We have demonstrated an increase in slow inactivation in the model of CM-iPSCs obtained from patients with a phenotype of cardiomyopathy combined with ventricular arrhythmias. Thus, this work contributes to understanding the role of the slow inactivation process in the mechanism of the development of heart pathology.
Objectives: The aim of this work was to study the effect of nanocluster polyoxometalate Mo72Fe30 on macrophages. In vitro studies of polyoxometalate’s effect on cultured macrophages allowed us to evaluate the reaction of immune cells (in particular, peritoneal and alveolar macrophages) to the nanoparticles. Results and discussion: The analysis of the obtained data allowed to establish that Mo72Fe30 is not toxic for peritoneal and alveolar macrophages, has no significant effect on the morphology of cells, as well as on the activity of α-naphthyl acetate esterase. Conclusion: However, polyoxometalate significantly reduces phagocytic activity, which may indicate possible polarization of macrophages. The obtained results confirm the possibility of using Mo72Fe30 in the field of biomedicine.
A new non-immortalized fibroblast-like cell line, named MSCWJ-3, was generated and characterized. Characteristics during long-term cultivation (6–24 passages) confirm the status of MSCs. It is shown: 1) a gradual increase in the proportion of senescent cells during long-term cultivation; 2) a significant decrease in the proliferation index by the 24th passage; 3) preservation of the normal diploid karyotype of the man (46, XY) during the entire period of cultivation, trisomy for different autosomes in single cells, absence of structural chromosomal rearrangements; 4) a high proportion of cells carrying surface antigens characteristic of MSCs: CD44, CD73, CD90, CD105, HLA-ABC and a low proportion with antigens CD34, CD45 and HLA-DR over 24 passages. Cells of the MSCWJ-3 line are capable of differentiation in the osteogenic and adipogenic directions at early and late passages; differentiation in the chondrogenic direction is absent. In general, there are some differences with previously obtained lines isolated from the same source and are associated mainly with the degree of expression of a number of status characteristics.
