The research work of the laboratory staff at the Institute of Experimental Cardiology began in 1979. The main focus of research until 1999 was the study of the β2-adrenoreceptor-dependent adenylyl cyclase system of lymphocytes in cardiovascular diseases, including dilated cardiomyopathy, primary pulmonary hypertension, arrhythmias and hypertensive disease.
Since 2000, the laboratory has been studying the mechanisms of leukocyte adhesion and migration in foci of inflammation, developing and testing peptide agonists and antagonists of chemotactic cytokines, and studying the role of immune cells in cardiovascular diseases.
The most significant results
• The mechanisms of leukocyte adhesion and migration on various types of matrix have been studied in vitro. It was shown that Mac-1 integrins (CD11b/ CD18) are involved in cell adhesion on fibrinogen/fibrin, while p150,95 (CD11c/CD18) integrins participate in migration stimulated by monocytic chemotactic protein-1 (MCP-1). Proteolysis of the fibrin(ogen) matrix leads to an increase in cell chemotactic ability.
• The intracellular signaling cascades activated by MCP-1 and participating in the migration of monocytes and endothelial cells in the concentration gradient of this chemokine were studied. Among the signal molecules, targets have been selected whose inhibition selectively impairs the migration of monocytes.
• Synthetic peptide fragments of MCP-1: peptide X (65-76), inhibiting MCP-1-mediated cell migration, and peptide IX, stimulating monocyte motility, were generated. Peptide X has been found to inhibit leukocyte migration in mouse air pouch model, inhibit subcutaneous endotoxin-induced inflammation in rats and primates, and slow down neointimal growth after balloon damage of rat carotid arteries. According to biosensor analysis (performed by colleagues from Prokhorov General Physics Institute of the Russian Academy of Sciences) the anti-inflammatory properties of peptide X are mediated by competition with chemokines for binding to glycosaminoglycan components of cell membrane and matrix glycoproteins. In patients with coronary artery disease the addition of peptide X (Ingramon) infusions to standard therapy before and after stenting of the coronary arteries, blood concentrations of C-reactive protein, fibrinogen and MCP-1 after the intervention were lower than in patients treated with standard therapy only. In experimental animals locally administered peptide IX caused the recruitment of monocytes, stimulated the formation of granulation tissue and angiogenesis, and increased the rate of wound healing. In collaboration with the researches from the Department of Physiology, Faculty of Medicine, Lomonosov Moscow State University, the cardioprotective effect of peptide IX was shown in the model of experimental myocardial ischemia-reperfusion.
• In patients with stable coronary artery disease (CAD), a negative correlation between the ratio of circulating regulatory T-lymphocytes to T-helpers 17 (Treg/Th17 ratio) and the severity of coronary and carotid atherosclerosis was observed. An increased number of producing INFγ Th17 contriubuted to the premature development of stenotic coronary artery atherosclerosis and CAD in men regardless of other risk factors. In a prospective study the prognostic significance of immunological parameters — the levels of circulating Treg, IL-17- and IL-10- producing T-lymphocytes — has been proven in identifying patients with rapidly progressing atherosclerosis of the coronary and carotid arteries.
• The effect of certain pharmacological agents on the parameters of cellular immunity was studied. Implantation of sirolimus-coated stents has been shown to result in a temporary increase in the relative amount of Tregs in blood. The mean quantaties of circulating Tregs were higher in patients receiving statins (atorvastatin), and an increase in the dose of atorvastatin was accompanied by a further expansion of the Treg population. These changes were not observed in patients receiving rosuvastatin.
• It was revealed that the contribution of immuno-inflammatory mechanisms to the development of atherosclerosis may depend on the patient’s age. In patients with coronary atherosclerosis, a decrease in the number of circulating Tregs is observed with age, while the level of effector T-cells was unchanged. The development of coronary artery lesions was associated with the decreased number of classical monocytes and the increased number of intermediate forms, which are the most inflammatory active cells. The relative amount of non-classical and intermediate monocytes increased with age.
• The content of monocytes with an "intermediate" phenotype and their complexes with platelets correlated with the severity of atherosclerotic lesions of the coronary arteries.
• In the culture of CD4+ lymphocytes “lipophilic” atorvastatin was shown to suppress the proliferation of effector cells more effectively than “hydrophilic” rosuvastatin, leading to an increase in the relative content of Tregs. Cultivation of lymphocytes in the presence of statins resulted in suppression of cell migration ability. In the culture of macrophages derived from human blood monocytes, statins inhibited M1-directed cell differentiation in a mevalonate-dependent way (the effect of “lipophilic” atorvastatin was more pronounced compared to “hydrophilic” rosuvastatin).
• The contribution of neutrophil granulocytes to atherogenesis is currently being studied.
• In patients with HFpEF a relationship between the severity of diastolic dysfunction and a decrease in the ratio of regulatory T-cells/activated T-helpers, and the number of blood monocytes.
Employees of the laboratory received the following patents for inventions:
- Peptide with the ability to inhibit monocyte cell migration stimulated by protein МСР-1. Invention patent 2260598, 20.09.2005.
- Peptide for inhibiting fractalkin-stimulated monocytic cell migration. Invention patent 2461565, 20.09.2012.
- Method for diagnosis of predisposition to atherosclerosis progression in patients with CHD according to the ratio of interleukin-10 and interleukin-17 concentrations in peripheral blood. Invention patent2566288, 24.09.2015.
- Method for diagnosis of predisposition to progression of atherosclerosis in patients with chronic coronary heart disease by the content of interleukin-10-producing T-lymphocytes in peripheral blood. Invention patent 2575791, 28.01.2016.
- Peptide with the ability to inhibit TARC-stimulated cell migration. Invention patent 2629198, 28.08.2017
- Method for diagnosis of predisposition to progression of carotid atherosclerosis in persons with normolipidemia by the content of CD4+ T-lymphocytes in peripheral blood. Invention patent 2636238, 21.11.2017.
- Method for prediction of restenosis development in patients with stable angina after coronary stenting. Patent for the invention 2724279, 22.06.2020.
The research work of the Cellular Immunology Laboratory is carried out in conjunction with the Departments of the Institute of Clinical Cardiology- Chronic IHD (since 2017 - the department of pulmonary hypertension and other heart diseases), new diagnostic methods, angiology, outpatient diagnostic and treatment technologies, the laboratories of atherosclerosis and peptide synthesis of Institute of Experimental Cardiology.
- Cell biology and immunology methods: flow cytometry, fluorescence and light microscopy, ELISA, immunohistochemistry
- Biochemical methods: electrophoresis, immunoblotting
- Primary cell culture and immortalized cell lines
- Аnimal models of human diseases
Arefieva T.I., D.Sc. – The Head of the Lab. eLIBRARY ID 84540.
Radukhina N.V., PhD - senior research fellow. eLIBRARY ID 155358.
Ruleva N.Yu., PhD – senior research fellow. eLIBRARY ID 49300.
Filatova A.Yu., PhD – research fellow. eLIBRARY ID 968936.
Ogurtsova E.S. – research laboratory assistant, a student of Lomonosov MSU, Faculty of Basic Medicine.
Researchers of the Lab actively participate with oral and poster presentations in annual scientific conferences.