We performed a simultaneous analysis of cytokine expression and metabolic reprogramming of macrophages upon combined stimulation of NOD1 and TLR4 receptors of innate immunity. NOD1 and TLR4 agonists boosted main parameters of glycolysis (extracellular acidification rate, glucose consumption, lactate release). However, changes of these parameters upon combined stimulation were not greater than those induced by stimulation of each individual receptor. At the same time, combined stimulation synergistically induced pro-inflammatory cytokine production and mRNA expression at relatively late time points (4–9 hours) after addition of agonists. In all, metabolic reprogramming may support synergistic induction of cytokines upon combined NOD1 and TLR4 stimulation; however, the origin of this synergy is in the synergistic induction of cytokine gene expression.

We provide the fi rst characterization of glucose and energy metabolism rearrangements in human macrophages upon their activation with a NOD1 receptor agonist (N-acetyl-D-muramyl-L-alanylD-isoglutamyl-meso-diaminopimelic acid, or M-triDAP) in comparison with the eff ects of a TLR4 agonist, lipopolysaccharide (LPS). We demonstrate possibilities of modulation of cytokine production by macrophages using glycolysis inhibitors.

Исследование нейроиммунных взаимодействий является одним из наиболее развивающихся направлений в изучении патогенеза рассеянного склероза. Механизмы этого взаимодействия до конца не ясны. Полагают, что ключевое значение в регуляции этого взаимодействия может принадлежать нейротрансмиттерам. Наибольшее внимание привлекают катехоламины, в частности, дофамин и норадреналин, рецепторы к которым экспрессируют клетки как нервной, так иммунной систем. Установлено, что модулируя функции иммунокомпетентных клеток дофамин и норадреналин способны влиять на течение как экспериментального аутоиммунного энцефаломиелита, так и рассеянного склероза. В работе представлен обзор литературы и собственных данных о значении дофамина и норадреналина в регуляции взаимодействия нервной и иммунной систем при рассеянном склерозе. Обсуждаются возможные механизмы, опосредующие влияние дофамина и норадреналина на патогенез рассеянного склероза, в частности, влияние дофамина и норадреналина на функционирование Th17-клеток, а также на опосредованный дендритными клетками Th17-зависимый иммунный ответ, играющий одну из ключевых патогенетических ролей при рассеянном склерозе. The neuroimmune interaction is one of fast developing directions in studying the pathogenesis of multiple sclerosis. The mechanism of this interaction is not sufficiently understood. The key role in regulation of this interaction is assumed to belong to neurotransmitters, among which catecholamines, specifically dopamine and norepinephrine, attract the greatest attention. Cells of both nervous and immune systems express dopaminergic and noradrenergic receptors. Dopamine and norepinephrine can influence the course of experimental autoimmune encephalomyelitis and multiple sclerosis by modulating functions of immune cells. This review presents literature and authors’ own data on the role of dopamine and norepinephrine in regulation of the nervous and immune system interaction in multiple sclerosis and focuses on possible mechanisms mediating the effect of dopamine and norepinephrine on the pathogenesis of multiple sclerosis, particularly the effect of dopamine and norepinephrine on the Th17 cell function and the dendritic cell-mediated Th17 immune response that plays a key role in the pathogenesis of multiple sclerosis.

This review presents data on the main functions of dendritic cells (DC) and their structure and stages of development. The role of DC in maintaining immunological tolerance is discussed, as it their role in the development of autoimmune diseases. The involvement of DC in the immunopathogenesis of multiple sclerosis (MS) is considered, along with their therapeutic potential in these cases.

Objective. To study the possibly associations between the clinical features of multiple sclerosis(MS) and quantitative and the qualitative properties of Th17 cells and the dopamine (DA) and serum noradrenaline (NA) concentrations in patients with MS. Materials and methods. Complex neurological and immunological investigations were carried out on 43 patients with remitting MS. All patients were underwent neurological investigations including assessment of levels of disability on the EDSS scale. Serum DA and NA concentrations were determined by immunoenzyme analysis. The proportions of circulating Th-17 cells were assessed by multicolor flow cytometry. The functional activity of Th17- and Th-1 cells was assessed in terms of IL-17 and γ-interferon production by peripheral blood mononuclear cells stimulated with magnetic particles coated with anti-CD3/anti-CD28 antibodies. Results. The proportion of Th-17 cells and cytokine production in the group of MS patients in exacerbation were significantly greater than in remission and in the control group, while the DA level was lower. NA levels in MS patients were identical in exacerbation and remission but were significantly lower than in the control group. Conclusions. These data suggest that catecholamines have an inhibitory effect on Th17 cells in MS.

For the first time we carried out a clinical assessment of the safety, tolerability and clinical efficacy course of repeated administration of experimental modified autologous vaccine interleykin (IL-10) dendritic cells in two patients with secondary-progressive multiple sclerosis patient and one with relapsing-remitting multiple sclerosis. In the course of treatment, we carried out clinical and immunological monitoring. It was found out that intradermal dose of 3 x 10⁶ cells applied to spinal area 6-12. times did not cause any serious side effects. After the treatment with dendritic cells, the following results were observed: 1) a significant positive clinical effect in patients with secondary-progressive multiple sclerosis exacerbations; 2) moderate positive clinical effect in patients with relapsing-remitting multiple sclerosis, in a state of remission; 3) a complete absence of any clinical results in patients with secondary-progressive multiple sclerosis without exacerbations. The immune response was characterized by a significant absolute and relative increase of serum T-regulatory cells. Discovered distinct anti-inflammatory properties of dendritic cell therapy allow us to consider it as a promising area of personalized treatment based on an individual vaccination against multiple sclerosis.

43 patients with remittent multiple sclerosis underwent complete neurological and immunological examination. All patients were examined using Expanded Disability Status Scale. Serotonin serum concentration was measured using enzyme immunoassay. Functional activity of Th1 and Th17 was assessed in accordance with IFNγ and IL-17 production by peripheral blood mononuclear cells. The study showed that cytokine production was significantly higher in patients with exacerbation of multiple sclerosis as in patients with remission or health volunteers. Serotonin level was comparable in all groups. Serotonin at final concentration 10-4 М inhibited IL-17 production.

AIM:Characterize the role of humoral immune response in mechanisms of action of muramyl dipeptide immune stimulators.MATERIALS AND METHODS:Mice were immunized by a complex of muramyl peptides (CMP) obtained from Salmonella typhi peptidoglycan and consisting of 3 components: 1) N-acetyl-D-glucoasminyl-(beta1- > 4)-N-acetyl-D-muramoyl-L-alanyl-D-isoglutaminyl-meso-diaminopimelic acid (GMtri); 2) N-acetyl-D-glucosaminyl-(beta1- > 4)-N-acetyl-D-muramoyl-L-alanyl-D-isoglutaminyl-meso-diaminopimeloyl-D-alanine (GMtetra) and 3) GMtetra dimer (diGMtetra), in which monomeric residues of GMtetra are linked by an amid bond between carboxyl group of terminal D-alanine of one of GMtetra residues and omega-amino group of meso-diaminopimelic acid of the other GMtetra residue.RESULTS:Immunization resulted in a multifold increase of IgM, IgG1 and IgG2a titers against CMP. Antibodies were directed against the whole molecule of diGMtetra and did not recognize its fragments. Sera of mice immunized with CMP protected the mice from lethal infection with Gram-negative (S. typhimurium) but not Gram-positive (Staphylococcus aureus) microorganisms.CONCLUSION:Induction of protective antibodies may present a novel mechanism of action of muramyl dipeptide immune stimulators.

The circulating CD4+ cytotoxic T lymphocytes (CD4+ CTL) represent an atypical subpopulation of + T cells that possesses some features of + T lymphocytes (lytic granules, expression of cytotoxic proteins, cell-mediated cytotoxicity). Here we show that CD4+ CTL, as compared to ordinary + T cells, display an elevated expression of RUNX3 and eomesodermin, two transcription factors that are vital for differentiation of + T lymphocytes and for the maintenance of their phenotype. This is accompanied by the reduced expression of ZBTB7B, a transcription factor playing a key role in the differentiation of CD4+ T lymphocytes. These changes of transcription factors observed in the + CTL penetrate phenotypically as an elevated expression of perforin and granzyme B, diminished expression of and appearance of low expression on 20-30% of + CTL. Thus, differentiation of + CTL from ordinary + T cells may be driven by a partial switch of the transcriptional program from the CD4 type to the CD8 type.

Muramyl peptides are potent activators of innate immune cells but, for an unknown reason, are weak inducers of pro-inflammatory cytokines in dendritic cells (DCs). Here, we show that in DCs stimulated by glucosaminyl muramyl tripeptide (GMtri) isolated from S. typhi, elevation of mRNA coding for tumor necrosis factor (TNF) and several other pro-inflammatory cytokines and chemokines is more short-lived than in lipopolysaccharide (LPS)-stimulated DCs and in GMtrior LPS-stimulated macrophages (Mphi). In addition, TNF and CCL4 mRNAs in GMtri-stimulated DCs are translated less efficiently than in LPS-stimulated DCs or in GMtrior LPS-stimulated Mphi. We found that Mphi, but not DCs, secrete interleukin-1beta (IL-1β) within first 1-2 h of GMtri stimulation. Addition of recombinant IL-1β results in an increase of TNF production by GMtri-stimulated DCs up to the levels observed in LPS-stimulated DCs and in GMtrior LPS-stimulated Mphi. Thus, the weak production of TNF by GMtri-stimulated DCs is likely caused by their inability to secrete IL-1β, which is an important autocrine regulator of cellular responses to muramyl peptides.

The paper presents the results of an open-label, placebo-controlled phase II/III clinical trial of an immunomodulatory drug Polymuramyl in patients with purulent surgical infections. An experimental group of 30 patients received the standard treatment together with Polymuramyl in a dose of 200 mcg IM once daily for 5 days. A control group of 30 patients received the standard treatment only. Polymuramyl significantly accelerated resolution of inflammation, reduced time to normalization of the body temperature, reduced pain, sleep disturbances and general discomfort. The overall effectiveness of treatment was judged as good in 83,3% patients in the Polymuramyl group and only in 56,7% in the control group (p<0,05 in the x-square test). Only one adverse event was registered in the Polymuramyl group, which was not caused by the drug being tested and did not require medical intervention. The laboratory examination revealed that Polymuramyl stimulated the components of the immune system that are involved in the elimination of extracellular purulent bacteria. In conclusion, Polymuramyl can be used as an auxilliary treatment of purulent surgical infection.