Vol 26, No 3 (2023)

Cover Page

Full Issue


Role of passive and active myostimulation for the changing levels of some cytokines

Ahmetyanov R.R., Davydovа E.V., Sabiryanov A.R., Shcherbakova I.L.


Injury to the anterior cruciate ligament (ACL) of the knee joint is complicated by development of arthrogenic muscle inhibition due to disregulating afferent influences on the excitability of the spinal and supraspinal tracts. The aim of our work was to study electromyography parameters, and myokine levels in the course of myostimulation in traumatic ACL injury.

28 male athletes with traumatic ACL injuries participated in the study. On admission to the clinic, all patients underwent electromyographic examination of the injured limb by the means of Viking Quest EMG/ EP apparatus (Nicolet, USA). Some patients, 10 days before starting the surgical treatment, underwent passive electrical myostimulation (EMS) of the quadriceps femoris muscle using the INTELECT® Advanced device (Chattanooga (DJO), USA). Further on, all patients underwent arthroplasty using a Karl Storz arthroscope (Germany). In the postoperative period, during immobilization for 2-weeks, the patients received EMS. After removing the orthosis, the patients switched to active training. The cytokine levels were studied using ELISA reagent kits from Vector-Best, Novosibirsk (IL-6), or from Cloud-Clone Corp. (China) for TGF–β1 assays. Statistical processing of the material was carried out using the Statistica package. vers.10.0 (StatSoft Inc., USA).

The highest average amplitude (μV) was recorded by electromyography in healthy individuals. In patients of the main group, significantly lower values of the average amplitude were recorded. After a 10-day EMS, a significant increase to the reference values of healthy individuals was noted. In the postsurgical dynamics, EMG indicators without EMS treatment remained at the same low levels. Meanwhile, the values following EMS treatment were comparable with those in healthy individuals, thus reflecting a faster and better muscle recovery after injury. The levels of IL-6 and TGF-β1 cytokines (myokines) significantly exceeded the initial levels in the course of EMS. The biological significance of increased IL-6 levels during the muscle exercise may consist the activation of AMP kinase and/or phosphatidylinositol-3-kinase at the level of skeletal muscles thus providing more efficient supply of energy substrate to the muscles. TGF-β1 promotes fibroblast proliferation, thus increasing collagen content.

Passive and active EMS leads to an improvement in electromyography parameters, along with increased concentration of myokines (IL-6 and TGF-β1) in peripheral blood, thus promoting improvement of energy balance, increasing the anti-inflammatory and repair potential of the damaged tissues.

Russian Journal of Immunology. 2023;26(3):211-216
pages 211-216 views

TLR-9 (-1237)*T/C polymorphism in russian COVID-19 patients from the chelyabinsk region

Belyaeva S.V., Suslova T.A., Stashkevich D.C., Balandina S.E., mjakotina D.E., Milonchenko M.S.


In COVID-19, the clinical outcome depends on a wide range of factors, including genetic features. Among them, TLRs, the genes encoding the receptors of innate immune system are of particular interest since they play the key role in development of innate immune response. The present study concerns the newely identified allelic variants of the TLR-9 (-1237)*T/C gene in Russian residents from the Chelyabinsk Region who had COVID-19 complicated by the bilateral viral pneumonia. Polymorphic variants of TLR-9 (-1237)*T/ C were determined by polymerase chain reaction. It was found that, among the COVID-19 patients, a TLR-9 allele (-1237 C) with higher transcriptional activity was more common than in the control group (19.421% and 11.275%, respectively, p = 0.019), and its homozygous genotype TLR-9 (-1237)*C was not detected in the comparison group. TLR-9 allele (-1237)*T in the patients with COVID-19 was less common in comparison with the control group (80.579% and 88.725%, respectively, p = 0.019). Taking into account the differences in suggested TLR-9 expression in more severe COVID-19 patients, we compared distribution of TLR-9 (-1237)*T/ С allele polymorphism in the patients with different severity of COVID-19. In the group of patients with mild form, the TLR-9 (-1237)*T/T genotype was more common as compared with patients who had more severe clinical course. The differences were significant at the trend level when compared with patients with a medium-severity disease (86.364% and 66,000%, respectively; p = 0.076).

Russian Journal of Immunology. 2023;26(3):217-222
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Some results of riamilovir (triazavirine) usage in medical staff for prevention and treatment of COVID-19

Berdiugina O.V., Gusev E.Y.


Riamilovir, a drug registered in Russia (chemical formula: methylthionitrooxodihydrotriazolotriazinide sodium, trade name: Triazavirin) is a synthetic analogue of guanine and a drug of direct antiviral action. Currently, there are conflicting data regarding usage of riamilovir as a preventive and therapeutic agent in coronavirus infections. The purpose of this study was to analyze some results of riamilovir usage, both for prevention of the SARS-CoV-2 infection and for the treatment of COVID-19 during the first wave of the new coronavirus pandemic.

The analysis was based on a survey of 62 medical staff workers at a single medical institution in Ekaterinburg who was ill with COVID-19, being divided into 4 groups: (1) those who did not receive riamilovir (control), (2) persons who received riamilovir only as a disease prevention, (3) subjects who received the drug as a therapeutic agent, (4) those who received riamilovir before and during the disease.

The data concerning usage of riamilovir for the prevention of infection with the SARS-CoV-2 virus have shown the following consequences: increased duration of hospitalization, an increased incidence of COVID-19 complications, i.e., fever, shortness of breath, pulmonary insufficiency, pneumonia, higher frequency of neurological disorders, which were not reported elsewhere. Severe clinical course of the disease was observed much more often in cases of prophylactic riamilovir administration, and the rehabilitation period was incomplete 2 months after the disease. Clinical symptoms of muscle and joint pain were documented at later terms in all persons who received riamilovir to prevent a new coronavirus infection. Usage of riamilovir for therapeutic purposes made it possible to avoid the development of pulmonary insufficiency, severe course of the infectious disease, and entirely restore the state of health.

The study did not reveal the usefulness of riamilovir for prevention of COVID-19 complications when taking the drug before infection with the SARS-CoV-2 virus. However, the use of riamilovir for therapeutic purposes prevents development of severe clinical cases and is associated with 4-fold reduced risk of pain in muscles, joints, and spine among the COVID-19 patients.

Russian Journal of Immunology. 2023;26(3):223-232
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Studies on the hormone and cytokine producing function of human cumulus cells and its interrelation with fertility in polycystic ovarian syndrome

Valikova O.V., Zdor V.V., Tikhonov Y.N., Boroda A.V., Gracheva A.M., Kolbin K.G.


Cumulus cells (CC) are the specialized layer of follicular cells that are in close contact with the oocyte. They are considered as indirect markers of the oocyte quality. Changes of these cells suggest a damage of the ovary. Determination of cytokines in cumulus cell culture may predict the chance for the conception and development of pregnancy. The objective of the present study was to obtain a primary culture of CC from healthy donors and patients with polycystic ovarian syndrome (PCOS), to identify the most significant differences in production of key cytokines in the CC monocultures of patients and healthy individuals in order to predict the results of in vitro fertilization. Materials and methods: the cell culture technique was used, i.e., cumulus cells of healthy donors and patients with polycystic ovary syndrome were obtained by transvaginal puncture of follicles in the in vitro fertilization (IVF) program. This procedure does not affect the rights of embryo, since the CC are not used at the stage of IVF procedure and intracytoplasmic sperm injection (Ethical Committee Protocol No. 9 of May 16, 2022). IL-6, IL-10, IFNγ and progesterone parameters were tested in adhesive cultures of CC by ELISA technique on days 1, 3, 7 of in vitro experiments. Results: We revealed a continuous secretion of progesterone, IL-6, IL-10, IFNγ in adhesive monocultures of CC. In the patients with PCOS, we have found a sharp increase of progesterone level in cultural media (p < 0.01) on the 7th day, By contrast, the initially increased progesterone levels proved to be significantly decreased in donors on the 7th day of culture. Moreover, in the culture of CC from patients with PCOS (7th day of the experiment), the values of IL-6, IL-10 increased only two-fold compared with 30-fold increase of these cytokines in healthy donors (p < 0.01). At the same term, we have observed a threefold decrease in IFNγ in the CC cultures of PCOS patients (p1-7 < 0.01; p < 0.05) compared with cultured controls, which showed a 20-fold increase (p1-7 < 0.01), thus determining differences in total cytokine balance and, probably, influencing the pregnancy prognosis. Conclusion: Significant multidirectional changes of cytokine levels in the culture of cumulus cells of the patients with PCOS and in healthy individuals may be regarded as determining factors in formation of blastocyst and preservation of the embryo. A further in vitro research on the production of cytokines and sex steroids by CC is especially important on day 5 to 7, when the oocytes are selected for entry into the in vitro fertilization cycle. The study of morpho-functional properties of little-studied cumulus cells using the cell culture technique will enable us for a deeper study on the mechanisms of disturbed folliculogenesis in PCOS, and, thereby, improve the reproductive prognosis in this disorder.

Russian Journal of Immunology. 2023;26(3):233-240
pages 233-240 views

Disturbances in the b cell component of immune system and associated immune alterations in post-covid patients

Dobrynina M.A., Zurochka A.V., Komelkova M.V., Zurochka V.A., Sarapultsev A.P.


There is only limited data on B-cell response in post-COVID patients despite its importance in studying the post-infection immunity. The present study aimed to investigate the features of the B-cell response in post-COVID patients, focusing on various B cell phenotypes. Along with the standard immunogram, the following cell phenotypes were examined: common B cells (CD45+, CD3-, CD19+); common memory cells (CD45+, CD3-, CD19+, CD27+); common non-memory cells (CD45+, CD3-, CD19+, CD5+); B1 memory cells (CD45+, CD3-, CD19+, CD5+, CD27+); B2 memory cells (CD45+, CD3-, CD19+, CD5-, CD27+); B1 non-memory cells (CD45+, CD3-, CD19+, CD5+, CD27-); and B2 non-memory cells (CD45+, CD3-, CD19+, CD5-, CD27-). The study revealed a sharp increase in B1 memory cells in 15.3% of post-COVID patients with impaired levels of B1 memory cells. This increase was accompanied by elevated levels of total B memory cells, B1 total lymphocytes (mainly, B1 memory cells), total T lymphocytes, and total IgA. By contrast, the patients with impaired B1 memory cells exhibited a sharp decrease in plasma cells, B2 lymphocytes (both memory and non-memory cells), natural killer cells, T regulatory cells, early activation T cells (CD25+), and C3a complement fragment. These findings suggest a unique immune system disorder characterized by dysregulated B lymphocyte switching from IgM to IgG and IgA synthesis, thus resulting in marked decrease in B2 lymphocyte subpopulations. This disorder may be associated with reduced T regulatory lymphocytes and early activation of T lymphocytes responsible for regulating B lymphocyte differentiation. Furthermore, the patients also exhibited reduced hemoglobin and platelet parameters, thus, potentially, contributing to hypoxia and blood clotting abnormalities. Thus, the phenotype identification of these immune system disorders in post-COVID patients requires non-standard approaches to assessing immune status, thus compicating clinical examination, but highlighting the need for immunocorrective therapies. These findings contribute to better understanding of post-COVID immune system disorders and require further investigation into the underlying causal factors.

Russian Journal of Immunology. 2023;26(3):241-250
pages 241-250 views

Significane of determining antinuclear antibodies in systemic connective tissue disorders in children

Zhuzhula A.A., Kurbatova O.V., Snovskaya M.A., Petrichuk S.V., Komyagina T.M., Tryapochkina A.S.


Systemic connective tissue diseases (SCTD) are characterized by systemic autoimmune inflammation and are accompanied by development of antinuclear antibodies (ANA). Our aim was a comparative analysis of ANA in blood serum in children with systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). The study included 50 patients with SLE, 50 patients with RA who were treated at the National Medical Research Center of Children’s Health. Median age was 12,2 y. o. (9-15.5). The titers of ANA and the cell fluorescence type were determined with the indirect immunofluorescence reaction (IFR) using the HEp-2 cell line (Immco Diagnostics, Inc, USA), as well as the concentration of antibodies to double-stranded DNA (adsDNA) in blood serum samples of the children detected by immunochemiluminescence (ICM) woth Elia™ dsDNA (Thermo Fisher Scientific, USA). A positive ANA titer and adsDNA were found, respectively, in 98% and in 48% in children with SLE. A positive ANA titer and adsDNA was detected in 100% and in 4% of children with RA, respectively. Highly positive ANA titers (> 1/1280) have been detected in 68% of children with SLE, and in 30% of children with RA. None of the RA patients with highly positive ANA titers had adsDNA. But, in patients with SLE, highly positive ANA titers and a positive dsDNA level were simultaneously detected in 16% of cases. There are both single types of cell fluorescence and their combinations in children with SLE and RA. Nuclear dot-like fluorescence was more common in children with SLE, cytoplasmic type, in children with RA, nucleolar type of glow was found only in children with RA. The revealed combinations of ANA and adsDNA titers in children with SLE and RA confirm the need for simultaneous use of RNIF and ICM.

Russian Journal of Immunology. 2023;26(3):251-258
pages 251-258 views

T helper subsets during the acute post-traumatic period in children

Zakirov R.S., Petrichuk S.V., Freidlyn E.V., Kuptsova D.G., Yanyushkina O.G., Karaseva O.V.


Severe mechanical injury is among the main reasons of disability and mortality in pediatric patients. The imbalance between the states of inflammation and immune suppression during the critical period of post-traumatic injury bears an elevated risk for infectious complications and/or multiple organ failure. The present study aimed to determine the informative immunological criteria in order to evaluate severity and prognosis for clinical outcomes in children from the severe injury group (SInj, ISS ≥ 16, n = 87); mild/moderate injury group (MInj, ISS < 16, n = 34), as based on assessment of helper T cells (Th) ratios, i.e., Th17/Treg, T127hi/ Treg, and Th17/T127hi. The patients with severe injuries were classified by their outcomes (favorable, n = 47; unfavorable, n = 40), presence of infectious complications (IC, n = 16) and the development of multiple organ failure (MOF, n = 11). Th ratios were studied on the 1st, 3rd, 5th, 7th, 14th day after injury. For the Sinj group, a pronounced increase of Th17/Treg ratio in the acute post-traumatic period with a decrease by 14 days was revealed. The indices of T127hi/Treg ratio on the first day for the patients from Minj group corresponded to the values of control group and significantly differed from patients with SInj in the 3rd to 5th day. There are different kinetics of Th subset ratio in peripheral blood of children with severe injuries over time in different groups, as well as with/without MOF, in presence, or absence of infectious complications and different clinical outcomes. Significant differences in T127hi/Treg ratio level were found in group with IC from 1st to 3rd day and from 7th to 14th day. Significant differences in Th17/Treg ratio level were found in IC group (7th day), in MOF group on 14th day post-injury. The patients with MOF had lower median concentrations of Th17/Treg and T127hi/Treg than patients without MOF. The results of the study indicate that the levels of Th17/Treg and T127hi/Treg ratio in children may be used to predict outcome of the traumatic disease and assess the risk of infectious complications and multiple organ dysfunction syndrome.

Russian Journal of Immunology. 2023;26(3):265-272
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Immunopharmacological aspects of studying immunotropic properties of a novel biocompound

Zabokritskiy N.A.


The article presents an experimental study of immunotropic activity of a new compound, a metabiotic, based on metabolites (biologically active substances) produced by a of saprophytic Bacillus subtilis B-9909 strain, being safe for health and standardized at Russian National Collection of Industrial Microorganisms (RNCIM). The aim of our study was to experimentally evaluate the immunotropic effects of metabolites produced by the probiotic Bacillus subtilis B-9909 upon the parameters of cellular immunity tested in the animal model of induced toxic liver damage. The metabolites were isolated from the cultural liquid of Bacillus subtilis culture (RNCIM strain B-9909) during its deep cultivation in a medium containing hydrochloric acid hydrolyzate of soy flour, or pancreatic hydrolyzate of casein. At 16-18 hours of cultivation, the cells were in the exponential growth phase. The indices of cellular status were studied in experimental groups of animals, when assessing therapeutic efficacy of an experimental metabiotic sample, as compared to a group of laboratory animals treated with the reference drug (ursosan). We assessed the quantitative indicators of blood as follows: determination of phagocytic activity (FA) of peripheral blood neutrophils; measurement of metabolic activity in peripheral blood neutrophils by means of NBT-test; quantitative determination of T and B lymphocytes; the number of antibody-forming cells (AFC). Liver damage was studied in a model of acute toxic hepatitis in albino rats. Experimental toxic hepatitis was induced by 40% solution of CCl4 in vaseline oil injected intragastrically for 2 weeks at the dose of 0.2 g/kg. The results of experimental studies showed activation of phagocytic activity by peripheral blood neutrophils at the early terms of experiment, being also confirmed by the results of NBT-testing. Moreover, at the early observation terms, a significantly increased representation of T and B lymphocyte populations as well as AFC numbers, were revealed thus suggesting activation of all components of cellular immunity in response to the carbon tetrachloride toxicity. Thus, the reported studies on the cellular status of laboratory animals treated with metabolites produced by probiotic microorganisms of the genus Bacillus subtilis (B-9909) on laboratory animals with toxic liver damage allow us to conclude that the test sample of this metabiotic preparation exerts a significant immunomodulatory effect, when compared with ursosan. This evidence allows us to consider this compound a promising candidate drug for a new hepatoprotector with immunotropic effect.

Russian Journal of Immunology. 2023;26(3):259-264
pages 259-264 views

Phases of systemic inflammation in septic and haemorrhagic shocks

Solmatina L.V., Zotova N.V., Zhuravleva Y.A., Brazhnikov A.Y., Gusev E.Y.


Shocks of different origin (both septic and aseptic) be be considered clinical equivalents of systemic inflammation (SI) with following main manifestations : pronounced hypercytokinemia, other markers of systemic inflammatory response (SIR), coagulopathy, multiple organ failure (MOF), hypothalamic-pituitary-adrenal (HPA) distress, systemic tissue alteration. In general, these phenomena are directly and inversely related to systemic microcirculatory disturbances which determine the pathogenesis of distinct shock states. The aim of our study was to identify the features of SI phases in the development of two variants of septic shock, i.e., acute course (first week of the process) and prolonged/subacute sepsis (2 to 6 weeks from the onset of manifestations), as well as haemorrhagic shock studied 4-12 hours from the onset of massive blood loss. To verify the SI phases, we used the previously proposed SI scale, including the value of six SIR levels (RL-0-5), as well as additional criteria of SI, i.e., evaluation of clinical MOF grade (SOFA scale), plasma D-dimers (> 500 ng/mL), cortisol (> 1380 nmol/mL), tissue alteration markers, e.g., myoglobin (> 800 ng/mL) and/or troponin I (> 0.2 ng/mL). To calculate RL in SIR, plasma CRP and four cytokines (IL-6, IL-8, IL-10, TNFα) were determined. The presence of SI was established if the SI scale exceeded 5 points (numerical RL values + presence of additional criteria, each equal to 1 point). The time and severity of the developing critical state, as well as the RL scores, were taken into account when reviewing the phases.

There were three main SI phases: (1) evolving condition, (2) cytokine storm/phlogogenic hit (SD-4-5), and depressive (exhausting) phase. The latter was characterized by relatively low SIR values (RL-2-3). The lethality rate for shock in the presence of acute sepsis (n = 13) was 71.4%, reaching 94.1% in prolonged sepsis (n = 17). For haemorrhagic shock after massive blood loss, if not resolved within 24 hoursm the mortality rates was 53.8% (n = 13). Development of shock in acute sepsis, and haemorrhagic shock (within 4 to 12 hours after the onset of massive blood loss) is accompanied by the severity of critical-phase cytokine storm/PPS with a predominance of RL-5 in cases of lethal outcomes, and by a depression phase (RL-2-3) in prolonged sepsis. Overall mortality (for all groups) was 66.7% in the PS phase, 89.5% in the depressive phase and 15.4% in the evolvingl phase of haemorrhagic shock (with possible transition from this phase to the more critical SI phases until lethal outcome).

Shock states of both septic and aseptic origin are based on a typical pathological process of SI, which should be distinguished from the signs of SIR characteristic of high-intensity canonical inflammation. It is characterized by higher cytokinemia (cytokine storm phase) or by the presence of additional SI phenomena with relatively moderate SIR levels (depressive SI phase). Thus, the depressive phase of SI is more fatal for disease outcome compared to the cytokine storm phase with higher intensity of SIR.

Russian Journal of Immunology. 2023;26(3):273-280
pages 273-280 views

Combined effect of methotrexate and bifidobacteria metabolites on TNFα AND IFNγ production by human peripheral blood mononuclears

Ivanova E.V., Chaynikova I.N., Bekpergenova A.V., Bondarenko T.A., Chelpachenko O.E., Zdvizhkova I.A., Perunova N.B., Bukharin O.V.


Methotrexate (Mtx) is a first-line drug for the treatment of numerous rheumatic and non-rheumatic disorders, including oncological disdiseases. However, therapeutic efficacy of Mtx is limited by severe toxicity to many organs (myelo-, hepato-, nephrotoxicity, mucositis, enteritis, dysbiosis at various human biotopes, etc.). Recently, a number of studies showed that some metabolites of Bifidobacteria and Lactobacilli are able to enhance effect of chemotherapeutic drugs and limit their toxic properties. The aim of the present work was to study the possible potentiating action of Bifidobacteria cell-free supernatants and methotrexate upon secretion of pro-inflammatory TNFα and IFNγ cytokines by human peripheral blood mononuclear cells (PBMCs). The immunoregulatory effects upon production of TNFα and IFNg  was evaluated in the in vitro model of cultured PBMC supplemented with Bifidobacteria metabolites, methotrexate, or their combination. Analysis of the combined effect of Bifidobacteria metabolites and Mtx on the cytokine production revealed their synergism towards the key pro-inflammatory cytokines (TNFα and IFNγ). We found an increase against the control cultures (with Mtx only), inhibition of the early pro-inflammatory cytokine TNFα production. On the contrary, we revealed an increased secretion of IFNγ which regulates the effector cells. The results obtained with these cytokines suggest the presence of a potentiating effect of Bifidobacteria metabolites upon anti-inflammatory and immunoregulatory properties of methotrexate. Thus, Bifidobacteria metabolites can be considered a promising agent which potentiates the therapeutic action of methotrexate by suppressing TNFα secretion and stimulating IFNγ by immunocompetent cells. Further studies of the combined effects of Mtx and metabolites from the intestinal microbiota upon the cytokine production by effector cells could be recommended, aiming to enhance therapeutic effect of methotrexate and limit its toxic properties using the Bifidobacteria metabolites.

Russian Journal of Immunology. 2023;26(3):281-286
pages 281-286 views

The effectiveness of immunomodulatory therapy with a synthetic analogue of the active center of the hormone thymus thymopoietin in the complex treatment of immunocompromised women with chronic infectious and inflammatory diseases of the pelvic organs

Kovaleva S.V., Nesterova I.V., Pikturno S.N., Chudilova G.A., Lomtatidze L.V., Teterin Y.V., Pirogova A.I., Prosolypova N.S., Chulkova А.M.


Chronic inflammatory diseases of the pelvic organs (pelvic inflammatory disease, PID) in women is among the main understudied problems in gynecology worldwide with adverse medical and socio-economic consequences, thus justifying the need for further study of immunopathogenesis and development of new approaches to treatment. Our objective was to develop new immunotherapeutic approaches to correction of combined disorders of the immune system functioning in immunocompromised women with PID and to evaluate their clinical and immunological efficacy.

55 women aged 20-40 years were examined, i.e., 35 patients with exacerbation of sluggish or recurrent PID, resistant to conventional therapy. Testiung was performed before complex treatment (study group 1 – GI- 1) and after the course (study group 2 – GI-2). Contents of T and B lymphocytes, natural killer cells (NK) (CYTOMICS FC500, USA), phagocytic and microbicidal functions of neutrophilic granulocytes (NG) were assessed in SG-1 and SG-2 before and 2-3 days after complex treatment with addition of an immunotherapeutic drug based on hexapeptide (HP) at a daily dose of 45 mg/ml intramuscularly for 10 days.

In patients from SG-1, a decrease in T cells (CD3+CD19- ) and B cells (CD3-CD19+), a 2-fold increase in the content of NK CD3-CD16+CD56+ was found, along with altered functioning of NG (deficiency of actively phagocytizing NG, a decrease in their digestive function and NADPH-oxidase activity). In SG-2 patients, the treatment was followed by restoration of the T (CD3+CD19-) and B cells (CD3-CD19+), NK cells (CD3-CD16+CD56+), like as an increase in effector functions, i.e., microbial capture by NG and their killing ability due to activation of NADPH oxidases and normalization of microbicidal reserve capacity in the NG cell population. Positive clinical effect included reduction of clinical symptoms in acute period, absence of PID exacerbations over follow-up for 6 months (85.6% of cases). Occasional exacerbations of PID were associated with medical manipulations (5.7%) and unprotected sexual contacts (5.7%).

The immunopathogenetically proven approach to correction of combined functional impairment of immune system in the women with PID shows a positive clinical and immunological effect.

Russian Journal of Immunology. 2023;26(3):287-294
pages 287-294 views

Cytokine profile in adolescent children with recurrent infections and polypous rhinosinusitis, ways of their correction

Korkmazov A.M., Kiseleva E.O., Frolova V.D., Romanyugo G.D.


A brief review of studies on one of the topical issues of otorhinolaryngology - polypous rhinosinusitis in people with recurrent infections is given. It was noted that patients with polyposis rhinosinusitis are more often committed to acute respiratory infections than those without nasal polyposis and they have a significant immunological imbalance. Serum levels of IL-8, IL-1β, anti-inflammatory IL-4 and IFN-α production were examined in 48 frequently ill patients with a history of respiratory allergopathology and polypous sinusitis. All patients, according to clinical recommendations, received complex conservative therapy. The control group consisted of 21 people who had respiratory infections no more than one or two times during the year and a history that was not aggravated by polypous rhinosinusitis. As a result, a significant decrease in IFN-α levels and an increase in the concentrations of IL-8, IL-1β, and IL-4 were revealed, which in turn confirms a violation of the body's immunoreactivity, a decrease in local and systemic immunity in patients with recurrent infections and polypous rhinosinusitis. The results obtained predetermine the need for the use of immunocorrective measures in the complex therapy of these patients. 

Russian Journal of Immunology. 2023;26(3):295-300
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Some immunological aspects of targeted therapy in polypous rhinosinusitis

Korkmazov M.Y., Lengina M.A., Dubinets I.D., Kravchenko A.Y., Klepikov S.V.


Over the past decade, targeted therapy with various monoclonal antibodies has become particularly relevant for the treatment of chronic polypous rhinosinusitis (PR). This is primarily due to the high incidence rate, polyetiological origin and pathogenetic features of polyposis development, low effectiveness of existing treatment approaches, the tendency for relapse, and comorbid conditions. The article provides a brief historical background concerning various predictors of the mucous membrane remodeling in the nasal cavity and paranasal sinuses at the stages of the polyposis formation thus justifying the need for implementation of monoclonal antibodies in the treatment schedules. Considering the leading role of Th2-inflammation in immunopathogenesis of developing polypous vegetations, the influence of targeted therapy upon treatment of chronic polypous rhinosinusitis is theoretically evaluated, and we highlight some important issues that should be further specified. Undoubtedly, inhibition of the synthesis of “necessary” interleukins leads to improvement in clinical symptoms and reduced size of polypous vegetations. At the same time, the real biochemical transformations of the nasal mucosa have been scarcely studied. E.g., an attempt to inhibit some cytokine may lead to indirect blockage of other pro-inflammatory cytokines. In future, it is necessary to study the pharmacodynamics of targeted drugs in order to clarify distinct contraindications to their use.

Russian Journal of Immunology. 2023;26(3):301-306
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Study of oxidative phosphorylation and glycolysis in CD4+T lymphocytes of HIV/HCV coinfected immunological non-responders by means of the seahorse technology

Korolevskaya L.B., Vlasova V.V., Shmagel N.G., Saidakova E.V.


Oxidative phosphorylation and glycolysis are essential for CD4+ T-lymphocyte survival, division, and functioning. However, indirect evidence suggests that in HIV-positive hepatitis C virus (HCV) coinfected immunological non-responders to antiretroviral therapy, the CD4+ T-cell metabolic activity parameters are violated. This information implies that in immunological non-responders, CD4+ T-lymphocytes' inability to productively divide and increase in number after viral suppression by antiretroviral drugs may be due to metabolic dysfunction. The newly released technology for the analysis of extracellular fluxes using seahorse XF equipment permits assessment of the cells’ metabolic activity. The aim of this study was to evaluate the efficiency of oxidative phosphorylation and glycolysis in CD4+ T-lymphocytes of HIV/HCV coinfected immunological non-responders using Seahorse technology. Peripheral blood samples from patients of two groups were studied: HIV/HCV coinfected immunological non-responders with CD4+ T-lymphocyte count less than 350/µl and HIV/HCV coinfected immunological responders with CD4+ T-cell count more than 500/µl. In isolated CD4+ T-lymphocytes, the basal and maximal oxygen consumption rates by complexes of the mitochondrial electron transport chain, as well as the rate of medium acidification by protons formed during glycolysis, were assessed. It has been established that in HIV/HCV coinfected immunological non-responders, both basal and maximal oxygen consumption rates by CD4+ T-cell mitochondria are reduced. Moreover, in isolated CD4+ T-lymphocytes of immunological non-responders, the basal rate of glycolysis is increased. It can be assumed that a significant part of CD4+ T-lymphocytes in HIV/HCV coinfected immunological non-responders is activated and ready for homeostatic proliferation, which aggravates the need for additional energy and macromolecules. However, cells are unable to change their metabolism in a coordinated manner to meet these demands. The identified dysregulation of metabolic pathways may contribute to the low regenerative capacity of CD4+ T-lymphocytes in HIV/HCV coinfected immunological non-responders.

Russian Journal of Immunology. 2023;26(3):307-312
pages 307-312 views

Development of a model immunofiltration assay using a conjugate based on horseradish peroxidase

Kropaneva M.D., Khramtsov P.V., Bochkova M.S., Rayev M.B.


The aim of the study was to optimize the conditions for a model immunoassay in the immunofiltration format using diagnostic reagents based on horseradish peroxidase. Residual blood serum samples from patients in the “red” zone with a verified diagnosis of a new coronavirus infection were used as positive sera, and blood sera obtained before 2019 were used as negative samples. The procedure of immunofiltration analysis was carried out using a pool of negative and positive blood sera. Studies were carried out to optimize the analysis procedure and increase the significant characteristics of the test. Results. It has been shown that the addition of sodium dodecyl sulfate to a final concentration of 50 μM in the substrate buffer makes it possible to achieve a higher analytical signal and a stable result 10 minutes after the end of the analysis procedure. Such conditions of immunofiltration analysis as dilutions of the diagnostic reagent, the volume of the introduced sample and the amount of the S-protein of the coronavirus applied to the nitrocellulose membrane were optimized. It has been determined that using immunofiltration analysis it is possible to detect antibodies against the coronavirus S-protein in a dilution of a serum sample of more than 1/1000. The results of immunofiltration analysis reproduce the results of ELISA.

Russian Journal of Immunology. 2023;26(3):313-320
pages 313-320 views

Effect of tetrapeptide Acetyl-(D-Lys)-Lys-Arg-Arg-amide on immunological and biochemical parameters of Wistar rats using passive smoking models

Kuzmicheva N.A., Mikhailova I.V., Filippova J.V., Smolyagin A., Livshic N.M., Miroshnichenko I.V.


Tetrapeptides, the homologues of adrenocorticotropic hormone fragment (15-18), play a special role among the bioregulators of the immune system. These compounds have cerebroprotective, neuroprotective and antioxidant properties. However, no available studies concerned the immunotropic properties of tetrapeptides in the models with exposure to xenobiotics, thus making such research quite relevant. Our study concerned the effects of tetrapeptide Acetyl-(D-Lys)-Lys-Arg-Arg-amide (laboratory code KK1) on the immunological and biochemical parameters of 72 female Wistar rats exposed to passive smoking. The experimental animals were fumigated with tobacco smoke for 8 hours. Synthetic peptide KK1 was administered intranasally at a dose of 40 µg/kg/day five times a day for 10 days. Cotinine was detected only in blood serum of rats from experimental groups, thus confirming a contribution of this tetrapeptide to the trend for normalization of some immunological parameters in experimental animals subjected to passive tobacco smoking, expressed as an increase in thymus mass and the number of splenocytes, and a decrease in the circulating immune complexes compared to the parameters of smoking rats of the group. We revealed that passive tobacco smoking in rats was accompanied by a general tendency to accumulation of iron, lead and nickel in peripheral blood. There was a marked increase in the concentration of cadmium, lead and cobalt in rats of the experimental group compared with the content of these trace elements in the liver of smoking animals injected with tetrapeptide KK1. The revealed shifts in immunological indices may be based, firstly, on hepatotoxic effect of ecotoxicants, The lymphoid lineage is mostly affected thus causing hypoplasia of the central and peripheral immunity organs. An evident sign of such pathology is a decreased cellularity of hematopoietic and lymphoid organs found in the present study. Secondly, the tobacco smoke components with prooxidant action may disrupt cellular redox homeostasis, causing damage to cell membranes, resulting in necrosis or apoptosis, thus explaining the revealed decrease in the number of thymocytes, splenic karyocytes and a decrease in the weight of organs. Thus, our results suggest usage of experimental passive smoking in order to evaluate efficiency of the tetrapeptides. Administration of Acetyl-(D-Lys)-Lys-Arg-Arg-amide peptide to the passively smoking rats is associated with tendency to normalize the mass of the thymus and spleen, the number of thymocytes and splenocytes, and a decrease in circulating immune complexes. Further studies are required to elucidate the effects of tetrapeptides upon the immune system.

Russian Journal of Immunology. 2023;26(3):321-328
pages 321-328 views

Role of myeloid-derived suppressor cells in prediction of the effectiveness of biologics in children with psoriasis

Kuptsova D.G., Radygina T.V., Freidlin E.V., Kurbatova O.V., Murashkin N.N., Petrichuk S.V.


Psoriasis is a chronic inflammatory skin disease characterized by increased proliferation of epidermal cells, impaired keratinization, and an inflammatory reaction in the dermis due to activation of T-lymphocytes and synthesis of proinflammatory cytokines. Pathophysiology of psoriasis is associated not only with activation of proinflammatory reactions, but also with decreased anti-inflammatory functions of immunosuppressive cells. It is known that Treg, Breg and MDSCs do not perform their classical homeostatic functions in psoriasis. In recent years, there have been more and more cases of developing resistance to ongoing therapy with genetically engineered biological drugs (GEBD) in childhood, requiring replacement or discontinuation of the drug. The aim of our work was to estimate the content of MDSCs subpopulations and their functional activity in the peripheral blood of children with psoriasis at different effectiveness of biotherapeutic drugs. We examined 110 children with psoriasis vulgaris before the appointment of biologics, at 16 and 52 weeks of therapy with adalimumab, etanercept and ustikinumab, aged 6 to 18 years. Сomparison group consisted of 34 healthy children. The effectiveness of therapy was assessed by the achievement of PASI 75 by one year of therapy. Contents of myeloid-derived suppressor cells (MDSCs) and their subpopulations, and the activity of arginase-1 were assessed by multicolor flow cytometry. An increased content of MDSCs was found in children with psoriasis against the comparison group (p = 0.000). Analysis of the effectiveness of biologics in children with psoriasis, according to PASI, showed a significant reduction in disease severity in the group of patients with good effect, both at week 16 of therapy (p = 0.000) and by one year (p = 0.017). In the group of patients with good effect of biological therapy, percentage of total MDSCs population was higher, both before start of treatment and by 52nd week of therapy (p < 0.01). Children with psoriasis showed increased immunosuppressive function of MDSCs by arginase-1 activity versus the comparison group (p = 0.000). The arginase-1 activity in patients with psoriasis at the stage of disease regression (PASI < 10) was significantly increased relative to children in progressive stage of psoriasis (PASI≥10; p = 0.001). Thus, the content of MDSCs and their suppressive activity in children with psoriasis is an informative efficiency predictor of the biological drugs. Fading of biotherapy effect after the induction course is accompanied by decreased number of MDSCs and their functional activity.

Russian Journal of Immunology. 2023;26(3):329-336
pages 329-336 views

Role of minor lymphocyte populations in development of liver fibrosis in children with glycogen storage disease

Kurbatova O.V., Petrichuk S.V., Movsisyan G.B., Kuptsova D.G., Radygina T.V., Anushenko A.O., Semikina E.L., Potapov A.S.


Glycogen storage disease (GSD) is a rare condition that changes the way the body uses and stores glycogen. Objective: to evaluate the content of small populations of lymphocytes and their ratios in children with hepatic forms of glycogen disease depending on the stage of liver fibrosis. 148 children with GSD at the age of Me=7,7 [3,9;11,8] were examined. The comparison group consisted of 54 healthy children. The stage of liver fibrosis was carried out on the FibroScan F502 device (EchoSence, France). Immunophenotyping of lymphocytes was performed on CYTOMICS FC500 (Beckman Coulter, USA). Indicators of lymphocyte populations were analyzed as a percentage of deviation from the age norm. In children with GSD, an increase in the degree of liver fibrosis was revealed from age (R=0.57). Treg content in children with GSD was at the lower limit of the age norm and did not depend on the stage of liver fibrosis. The content of Th17 and Thact lymphocytes was significantly higher than that of the comparison group at all stages of liver fibrosis, starting from stage F1. With an increase in the stage of liver fibrosis, there was an increase in the proportion of patients with Thact and Th17 content exceeding the upper limit of the normative values (PF0-F4=0.021 and PF0-F4=0.012, respectively). An increase in Th17/Treg and Thact/Treg ratios was revealed in patients with GSD relative to the comparison group in all age groups, the dynamics of Th17/Treg and Thact/Treg ratios was characterized by their increase with age. Analysis of indicators depending on the stage of liver fibrosis in children with GSD revealed a significant increase in the Thact/Treg ratio from stage F0 to stages F1, F2, F3 and F4 (PF0-F4=0.000). The Th17/Treg index increased from stage F0 to stages F1, F2, F3 (PF0-F3=0.000).

An increase in the content of Thact and Th17 lymphocytes, as well as Th17/Treg and Thact/Treg indices with an increase in the stage of liver fibrosis can be used as an additional tool in assessing fibrotic changes in the liver. Immunological indicators objectively reflect the severity of the patient's condition with hepatic forms of GSD.

Russian Journal of Immunology. 2023;26(3):337-344
pages 337-344 views

Characteristics of the phagocyte effector indexes and effect of local cytokine therapy on their parameters in inflammatory diseases of the maxillofacial region

Latyushina L.S., Malysheva L.Y., Piotrovich A.V., Berezhnaya E.S., Lapteva A.V.


Odontogenic inflammatory diseases and injuries take a leading place in the structure of surgical dental pathology. They often proceed with inflammatory complications. The aim of the present work was to analyze the effector indexes of phagocytic cells at the local area of inflammation (secretions from purulent wounds, tooth socket and mixed saliva), and to evaluate the effect of topical cytokine therapy on the studied parameters in patients with inflammatory dental diseases (chronic periodontitis, odontogenic phlegmon) complicated by mandibular fractures. As a part of a two-stage research, 236 people were comprehensively examined, divided into groups depending on their clinical disease and method of treatment. This group included 74 patients (classified as K 04.5 – K 04.9), divided into subgroups by the types of periodontitis (chronic fibrous, granulating and granulomatous form); 102 patients with phlegmon (classified as L03.2, K12.2), who, at the second stage of basic therapy, received topical cytokine treatment with betaleukin (recombinant IL- 1β) and roncoleukin (recombinant IL-2). Fifty patients with fractures of the lower jaw (classified as S 02.6) underwent immunotherapy with betaleukin at the second stage of treatment. Examination of patients with inflammatory dental diseases at the first stage of study and analysis of the effector indexes of neutrophilic granulocytes and macrophages from the local inflammatory foci revealed some immunological predictive signs of lacking efficiency of the inflammatory response, manifesting as aberrant functional activity of phagocytes, e.g., inhibition of all studied parameters along with sufficiently reduced functional reserve of phagocytic cells in the patients with phlegmon; high ability to produce reactive oxygen species with a decreased functional reserve of neutrophils in the patients with fractures, and signs of activated chronic inflammation in the patients with periapical lesions. The second stage of our research associated with topical therapy with recombinant cytokines and analysis of the obtained data, ebabled us to detect different effects of drugs on the studied parameters and register the integral effect of immunotherapy, which represents normalization of altered functional indexes. Hence, the data obtained may indicate that, in patients with inflammatory processes in the maxillofacial region, a higher adaptive effector potential of phagocytes was formed at the affected focus following the topical cytokine therapy.

Russian Journal of Immunology. 2023;26(3):345-354
pages 345-354 views

Dynamics of matrix metalloproteinases and their tissue inhibitors in patients with herpesvirus and papillomavirus infection

Nevezhkina T.A., Chernikova M.A., Markelova E.V., Tulupova M.S., Kostyshko A.V., Fedyanina L.N., Markova N.Y.


Sexually transmitted infections are of great importance for the proper reproductive function in women. Chronic inflammatory process leads to reproductive disorders. A special role in the chronic inflammatory process is attributed to papillomavirus (PVI) and herpetic infection. MMP-2 and MMP-9 enzymes cleave type 4 collagen which makes the scaffold of basement membranes and contributes to the separation of endothelial cells from the membranes, followed by their further migration and direct participation in angiogenesis thus affecting the growth of tumors, in particular cervical cancer. Tissue inhibitors of matrix metalloproteinases are known to limit the collagen breakdown. However, the imbalance of MMP and TIMP is accompanied by accumulation of extracellular matrix and increased risk for reproductive disorders. The aim of our study was to evaluate the dynamics of acute-phase proteins affecting the state of intercellular matrix (MMP-2, MMP-9 and MMP tissue inhibitors (type 1, 2) in blood serum of patients with PVI or coinfection of PVI and HSV before and after therapy with drugs exhibiting antiviral and immunomodulatory effects, i.e., a synthetic compound (Inosine pranobex) and vegetable substance (Solanum tuberosum). We have examined 141 patients with papillomavirus and herpetic infections treated with Inosine pranobex and Solanum tuderosum. Determination of MMP-2, MMP-9 and TIMP-1, TIMP-2 levels of in blood serum was carried out using specific reagents from R&D Diagnostics Inc. (USA). The drug therapy with active substances of Inosine pranobex and Solanum tuberosum was associated with positive dynamics of the level of MMP-2, MMP-9 and tissue inhibitors of types 1 and 2 in all groups under studies. However, Inosine Pranobex exerts more pronounced changes, especially in subgroups with viral coinfections.

Russian Journal of Immunology. 2023;26(3):355-362
pages 355-362 views

Quantitative and phenotypic transformation of CD16+IFNα/βR1-CD119+, CD16+IFNα/βR1+CD119- and CD16+IFNα/βR1+CD119+ neutrophil granulocytes subsets in patients with post-COVID syndrome

Nesterova I.V., Chudilova G.A., Atazhakhova M.G., Matushkina V.A., Kovaleva S.V., Chapurina V.N.


In patients who underwent COVID-19, various manifestations of post-COVID syndrome (PCS) are noted, causing the development of disorders accompanying severe viral infections, complicated by chronic fatigue syndrome (CFS) and severe cognitive disorders (CD). Studying the molecular mechanisms of these disorders in the system of neutrophilic granulocytes (NG) in patients with PCS associated with IFN production, receptor function of NG, in particular, their subsets expressing IFNα/βR, IFNγR(CD119), is relevant for the search for therapeutic strategies, restoration and enhancement of the innate immune response after COVID-19.

Our objective was to clarify the quantitative and phenotypic characteristics of certain subsets of neutrophil granulocytes, i.e., CD16+IFNα/βR1-CD119+, CD16+IFNα/βR1+CD119-, CD16+IFNα/βR1+CD119+, in peripheral blood of patients with post-COVID syndrome.

We have examined 39 patients (24-60 years old) with PCS 3 months after COVID-19 (study group 1, SG1). The comparison group (CG) included 30 volunteers examined over the pre-COVID period. Detection of herpesvirus infections (HSV1, EBV, HHV6, CMV) was carried out in scrapings from the tonsils and the posterior wall of the pharynx. To determine the severity of the clinical PCS symptoms, a questionnaire was used to assess its severity using a point scale. The content and phenotype of NG subsets CD16+IFNα/βR1-CD119+, CD16+IFNα/βR1+CD119-, CD16+IFNα/βR1+CD119+ were assessed by means of FC 500 (Beckman Coulter, USA).

In all patients of SG1, clinical manifestations of CFS and CD were revealed, at the average severity rates of 16.0 points (14.75-20.25). When detecting herpesvirus infections, 37.2% had only HSV1 infection; 62.8% of patients showed mixed infection (HSV1, EBV, HHV6), which exhibited more pronounced clinical symptoms. We have noted absence of CD16+IFNα/βR1+CD119+NG subset and phenotype transformation of CD16+IFNα/βR1-CD119+NG, CD16+IFNα/βR1+CD119-NG subsets. Increased density expression of CD16, IFNα/βR1, CD119 receptors was also found (p1-3 < 0.05) thus suggesting ability to accept the interferon signaling and response.

Reduced infectious burden in the post-COVID period and adequate functioning of the immune system, including the neuroimmunoendocrine regulation mechanisms, should contribute to the functional recovery of various organs, systems, thus neutralizing the PCS manifestations. Therefore, usage of recIFNα2b in combination with highly active antioxidants may contribute to development of protective immunity, prevention of acute respiratory viral infections, exacerbation of chronic infections, and restoration of the NG phenotypes followed by restoration of anti-infectious immune balance.

Russian Journal of Immunology. 2023;26(3):363-372
pages 363-372 views

Effect of synthetic analogue of ZP2 peptide, an active center of GM-CSF, upon antilysozyme activity of Candida

Pashinina O.A., Kartashova O.L., Pashkova T.M., Gritsenko V.A.


Our aim was to analyze the effects of the ZP2 peptide, a synthetic analogue of active center of GM-CSF, upon antilysozyme activity (ALA) of Candida.

32 vaginal isolates of Candida spp were used in the work. Five species have been isolated from the vaginal secretions of the conditionally healthy pregnant women taken within a screening program. To study the effect of ZP2 on the ALA of the Candida fungi, the fungal cells were co-cultured with a ZP2 solution in Saburo broth medium at 37 oC for 24 hours. Thereafter, ALA of fungi was determined by photometric method.

It was found that the ZP2 peptide caused a decreased expression of ALA of vaginal Candida spp. Isolates, i.e., loss of ALA in the isolates of C. glabrata, C. tropicalis, C. krusei and some isolates of C. albicans, as well as a decreased level of ALA in C. kefir and other C. albicans isolates. Thus, the synthetic analogue of ZP2 peptide (active center of GM-CSF) showed an inhibitory effect upon the antilysozyme activity of Candida.

Russian Journal of Immunology. 2023;26(3):373-376
pages 373-376 views

Effect of cucurbiturils on the numbers and differential counts of peripheral blood leukocytes in laboratory animals after in vivo parenteral administration

Pashkina E.A., Aktanova A.A., Kovalenko E.A., Kozlov V.A.


Immunological safety of nanoparticles is an urgent problem for development of drug delivery systems used as the basis for creating these systems. Cucurbit[n]urils (CB[n]) are molecular nanocontainers that can encapsulate various drugs and serve as the basis for delivery systems. Cucurbiturils are low-toxic compounds: under in vitro conditions, they exhibit weak immunomodulatory properties, without sufficient immunotoxicity. The aim of the present study was to evaluate the in vivo effects of cucurbiturils on the number and differential leukocyte counts in peripheral blood.

BALB/c mice aged 2-4 months were used in the work. For this study, cucurbiturils were diluted in phosphate-buffered saline and administered to laboratory animals intraperitoneally (three times a week).

When evaluating the effect of cucurbit[7]uril on blood parameters of the animals after three intraperitoneal injections weekly, no statistically significant changes were registered. However, at first administration, the animals showed a slight increase in relative number of lymphocytes after the first injection of CB[6], and an increased proportion of neutrophils after the first administration of CB[8]. Meanwhile, the proportions of lymphocytes and neutrophils were within normal ranges after the 2nd and 3rd injections of cucurbiturils, and did not show any significant differences against the controls. Moreover, the impact upon the subpopulation composition of peripheral blood lymphocytes in laboratory animals was assessed. After injection of CB[6], there was a decrease in T lymphocytes in the peripheral blood, along with increased proportion of CD19+B lymphocytes compared with the controls. CB[7] and CB[8] injections did not affect the subpopulation profile of peripheral blood lymphocytes.

We have found that intraperitoneal administration of CB[n] did not affect the blood parameters of laboratory animals, thus, probably suggesting the in vivo safety of these compounds. At the same time, CB[6] is able to exert a stimulating effect on humoral immunity by increasing relative contents of B lymphocytes.

Russian Journal of Immunology. 2023;26(3):377-380
pages 377-380 views

Content of myeloid-derived suppressor cells in autoimmune diseases in children

Radygina T.V., Kuptsova D.G., Petrichuk S.V., Potapov A.S., Murashkin N.N., Abdullaeva L.М., Kurbatova O.V., Tsvetkova V.S.


Myeloid-derived suppressor cells (MDSCs) play an important role in regulation of immune response. An increase in their number in adult patients with autoimmune diseases has been reported. G-MDSCs, M-MDSCs, and MDSCs(M-G-) at different stages of autoimmune disease may both activate T cell proliferation, leading to disease progression, or inhibit it, thus promoting Treg differentiation. Arginase-1 (Arg- 1) is an enzyme in MDSCs that reduces the concentration of arginine required for T lymphocyte proliferation. Our aim was to evaluate the content of MDSCs populations and functional activity of MDSCs in children with autoimmune diseases. 75 children with inflammatory bowel diseases (IBD), 60 children with multiple sclerosis (MS), 69 children with psoriasis (PS), 62 healthy age-matched children were included into the study. The content of MDSCs ((CD3, CD19, CD56, HLA-DR)-, CD11b+ and CD33+), subpopulations of MDSCs (M-MDSCs, G-MDSCs expressing CD14 and CD15), assessment of Arg-1 activity were performed by flow cytometry techniques. The content of MDSCs in patients with IBD, MS and PS was significantly higher than in the comparison group and depended on the state of exacerbation/remission. In exacerbation and remission of IBD, MS and PS, a significant increase of MDSCs was revealed when compared with healthy children; the highest values were found in children in exacerbation of MS (Me-3.5 (2.5-5.6) % MNC against Me-1.6 (0.9-2.5) % MNC, p < 0.001). In patients with MS, the content of G-MDSC, M-MDSC was significantly higher, and MDSC(M-G-) was lower than in healthy children. An increase in absolute amounts of G-MDSCs was shown in MS exacerbation compared to the disease remission state (p = 0.022). For patients with IBD, a significant increase in percentage of MDSCs and M-MDSCs (p = 0.014 and p = 0.045, respectively) was obtained in exacerbation of the disease relative to remission state. In patients with IBD, MS, and PS, a significant increase in Arg-1 activity in MDSCs was found, with a decreased number of MDSCs in patients in remission compared to exacerbation phase of the disease. In children with autoimmune diseases, an increase in the MDSC populations was found. The activity of arginase-1 in MDSCs is increased in remission, along with a decrease in their numbers.

Russian Journal of Immunology. 2023;26(3):381-388
pages 381-388 views

Features of T cell immunity depending on the content of natural killer cells in patients with acute coronary syndrome following COVID-19

Safronova E.A., Ryabova L.V.


We examined 65 men with unstable angina and acute myocardial infarction (acute coronary syndrome - ACS) from 40 to 65 years old who had previously had COVID-19 and 20 people with ACS who had not undergone COVID-19. All persons also had hypertension, and they required stenting of the coronary arteries within the next 3 days after admission to the hospital. From the immunological parameters by flow cytometry on the Navios cytofluorimeter (BeckmanCoulter, USA), according to the standardized technology for assessing the lymphocytic link of immunity [1], the following were determined: ), CD45+ CD3+ CD8+ (cytotoxic T-lymphocytes), CD45+CD3-CD19+ (B-lymphocytes), CD45+CD3+CD16+CD56+ (TNK cells), CD45+CD3-CD16+CD56+ (natural killer cells), CD45+ CD3+CD4+CD25+CD127- (T-regulatory cells), CD45+ CD3+CD4+CD25+ (T-lymphocytes - early activation), CD45+CD3+HLA-DR (T-lymphocytes - late activation). All patients were divided into groups depending on the content of NK cells (natural killer cells). Patients who have had COVID-19 have 3 phenotypes of disorders (decreased NK cell count, normal and increased), while non-survivors have 2 phenotypes (decreased NK cell count and normal). The most severe condition and severity of immune disorders were found in patients who had undergone COVID-19. In patients with acute coronary syndrome and COVID-19, predominantly with normal and elevated levels of NK cells, compared with ACS patients without COVID-19, a more severe course of the disease was observed - patients with acute myocardial infarction prevailed, they had a higher mortality rate, the duration of treatment was increased, and stent thrombosis was also more common. In persons with ACS and COVID-19 with elevated NK cells, the maximum decrease in the T-cell immunity was observed: T-lymphocytes of general, T-lymphocytes-helpers, T-cytotoxic lymphocytes, T-lymphocytes of early activation, T-regulatory cells in absolute numbers compared to other groups. The lowest immunoregulatory index and, at the same time, the maximum number of T-NK-lymphocytes were observed in persons who had undergone COVID-19 and had reduced NK cells. The minimum number of T-NK lymphocytes was recorded in patients with low NK cells who did not have COVID-19. Minimal T-lymphocytes (CD45+CD3+CD4+HLA-DR+) of late activation were found in people who recovered from COVID-19 with elevated and normal NK cells. The lowest number of late activation regulatory T cells was observed in patients who did not have COVID-19, but were vaccinated, and had a normal content of NK cells. The study also allows us to more clearly define the groups of patients with ACS who need additional immunocorrection.

Russian Journal of Immunology. 2023;26(3):389-396
pages 389-396 views

Indicators of immunological response to non-drug therapy of chronic adenoiditis

Solodovnic A.V., Vyalova T.A., Gabova E.V., Pozdnyakov P.A., Bushueva D.S.


The aim of our work was to study individual indices of immune response in children with chronic adenoiditis and evaluate their dynamics following low-frequency ultrasonic cavitation combined with photochromotherapy included into the treatment regimen. The study involved 104 patients 3 to 15 years old, divided into three groups: the 1st control group (n = 34) received standard treatment for 7 days; 2nd group (n = 37) received a supplementary low-frequency ultrasonic cavitation irrigation of pharyngeal tonsil combined with photochromotherapy on lymphoid tissue of the pharyngeal tonsil for 7 days; the 3rd group (n = 33) received only low-frequency ultrasonic cavitation irrigation treatment. Comparative estimation of clinical and immunological indicators (sIgA, IL-1, IL-6, IL-8, IL-10, TNFα) was performed prior to the therapy (day 0) and on the 7th day from the start of treatment. Before therapy, a decrease in the content of sIgA, IgA was revealed in all groups. In the second group, there was a statistically significant increase in the level of IgA after treatment, which suggests activation of local immunity factors. The dynamics of proinflammatory cytokines (IL-6, IL- 10) also indicates effectiveness of the drug treatment by reducing manifestations of the tissue immunological reactivity in pharyngeal tonsil. An increased number of anti-inflammatory IL-8 and IL- 10 cytokines could be considered a compensatory response to decreased level of proi-inflammatory cytokines.

As a result, the inclusion of low-frequency ultrasonic cavitation in combination with photochromotherapy into the complex treatment for chronic adenoiditis thus stabilizing the course of immune response.

Russian Journal of Immunology. 2023;26(3):397-402
pages 397-402 views

Effect of pregnancy-specific β1-glycoprotein on the expression of arginase-1 and indolamine-2,3-dioxygenase by myeloid-derived suppressor cells

Timganova V.P., Shardina K.Y., Gutina E.V.


Myeloid-derived suppressor cells (MDSC) - a population of immature cells of myeloid origin with inhibitory functions, mainly related to T lymphocytes. Normally, MDSC account for less than 1% of leukocytes in peripheral blood. The number of these cells increases during healthy pregnancy. However, MDSC have been shown to play a critical role in the maintenance of tumor growth and in autoimmune diseases.

Because MDSC are now considered important regulators of immunity, finding ways to manipulate their functions is important for the development of therapies for malignant and autoimmune diseases, as well as for pregnancy pathologies and post-transplant complications. The immunosuppressive mechanisms of these cells are mediated by their expression of the surface molecules CD73, ADAM17, PD -L1, the enzymes arginase 1 (Arg 1), inducible nitric oxide synthase (iNOS), and indoleamine 2,3-dioxygenase (IDO), reactive oxygen species, and the production of the anti-inflammatory cytokines IL -10 and TGF-β1.

Pregnancy-specific β1-glycoprotein (PSG) is a pregnancy glycoprotein that has immunomodulatory effects on natural (dendritic cells and macrophages) and adaptive (T cells) immunity cells. At the same time, the effect of PSG on MDSC has not been investigated so far. Since this glycoprotein has promising pharmacological applications, it is necessary to study not only the native variant of PSG but also its recombinant form.

Since the main function of MDSC is immunosuppression, the aim of our work was to evaluate one of its mechanisms, namely the intracellular expression of amino acid degradation enzymes Arg1 and IDO under the influence of native and recombinant PSG in vitro.

MDSC differentiation was performed from CD11b+ cells isolated from peripheral blood of healthy volunteers. Cells were cultured for 7 days with stepwise addition of GM-CSF, IL -1β, and LPS. Native (n) (1, 10, and 100 μg/mL) and recombinant (r) (1 and 10 μg/mL) PSG was added to the cultures three days before the end of incubation. The percentage of MDSCs (Lin- HLA-DR -CD11b+CD33+) intracellularly expressing Arg1 and IDO was determined by flow cytometry.

It was found that nPSG and rPSG did not alter the amount of Arg1-expressing MDSCs at all concentrations examined. However, at a concentration of 10 μg/mL, both types of proteins caused a statistically significant increase in the percentage of cells expressing IDO.

We have already established that nPSG and rPSG affect MDSC differentiation by increasing the proportion of these cells belonging to the monocytic subpopulation. However, now we can say that PSG, in addition, enhances the suppressive function of the studied cells.

The obtained data are novel and open perspectives for targeting myeloid suppressor cells to improve cellular technologies in science and medicine.

Russian Journal of Immunology. 2023;26(3):403-408
pages 403-408 views

Effect of graphene oxide nanoparticles on apoptosis of T-lymphocytes and Jurkat cells

Usanina D.I., Uzhviyuk S.V., Zamorina S.A.


Graphene and its derivatives are materials with unique physicochemical properties. A detailed study of these materials allows to consider them prospective biomedical agents for targeted drug and gene delivery, photothermal therapy of cancer, bioimaging, etc. However, this requires a comprehensive studies of their effects on the body tissues, including cells of the immune system.

The aim of our research was to stydy the effects of nanoparticles based on pegylated graphene oxide (GO) upon apoptosis of T lymphocytes derived from blood of healthy donors and Jurkat 5332 cell line. Comparison of these cells will extend our knowledge of the effects of nanomaterials on the cells, and to respond the question, what results obtained with continuous cell lines are valid for normal non-malignant cells. In this work, we used GO nanoparticles (100-200 nm, 1-5 ìm) coated with linear (LP-GO) and branched (BP-GO) polyethylene glycol (PEG). The cells were cultured for 24 hours at 37 °C and 5% CO2 with nanoparticles at concentrations of 5 and 25 ìg/mL. Viability and early and late apoptosis of incubated Jurkat cells and CD3+ cells from healthy donors were assessed by flow cytometry. It was found that the small nanoparticles coated with linear PEG at high concentrations (25 ìg/mL) could significantly reduce the number of live cells and increase the number of cells in late apoptosis. At the same time, large nanoparticles coated with branched PEG at high concentrations (25 ìg/mL) increased the percentage of T cells in early apoptosis.

Meanwhile, the GO nanoparticles at both concentrations did not affect the viability and apoptosis of Jurkat cells, regardless of the size, concentration, and type of surface function of the particles.

The obtained results suggest that GO nanoparticles exert different effects upon normal and malignant lymphocytes of T lineage. One may assume that these discrepancies could be explained by greater resistance of tumor cells compared to normal T cells. These findings suggests that studies of nanomaterials upon living cells should not be limited to experiments on cell lines, since their properties may significantly differ from those of non-malignant cells.

Russian Journal of Immunology. 2023;26(3):409-414
pages 409-414 views

Flow cytometry analysis of PD1 expression on rat blood CD3+ lymphocytes stimulated by CD3 antibodies

Khramova T.V., Beduleva L.V., Fomina K.V., Abisheva N.N.


The role of the PD-1/PD-L signaling pathway in the regulation of the immune response is currently the focus of research. Numerous studies have shown the key role of PD-1 molecules in the regulation of autoimmune, antitumor and antiviral reactions. The culture of rat and mice lymphocytes, as well as animal experimental models of immunopathologies are widely used in research. However, rat lymphocytes are almost not used to study the PD-1/PD-L pathway. There is no data on PD-1 expression or methods of its induction on rat lymphocytes. In human T-lymphocyte culture, PD-1 expression can be induced by NIB1412 anti-CD3 antibodies coated in the well of culture plates. In this study, we investigated the effect of G4.18 anti-CD3 antibody on PD-1 expression in peripheral blood lymphocyte of intact Wistar rats in vitro. According to some literature data, G4.18 anti-CD3 antibodies in immobilized form can activate isolated rat T cells, and according to others, inhibit allogeneic reactions in mixed lymphocyte culture and block cytotoxicity of cells obtained from rats with a developed graft rejection reaction. We found that incubation of rat blood lymphocytes with G4.18 anti-CD3 antibodies immobilized on plastic leads to a change in cell morphology and induction of PD-1 on CD3+ lymphocytes. After incubation with anti-CD3 antibodies, the proportion of PD-1+ lymphocytes among CD3+ lymphocytes was 12.05±6.04%, which is significantly higher than the proportion of such cells before incubation and during incubation in a cultural medium, which amounted to 2.60±2.62% and 4.59±5.81%, respectively. In the dot-plot graphs showing the distribution of cells according to the parameters of forward scatter and side scatter, PD-1+CD3+ lymphocytes induced by anti-CD3 antibodies are localized in the region of relatively lower forward scatter and greater side scatter. Perhaps these cells belong to apoptotic cells.

Russian Journal of Immunology. 2023;26(3):415-420
pages 415-420 views

Levels of tissue inhibitor of matrix metalloproteinases type 1, matrix metalloproteinase type 7 and anthropometric parameters in practically healthy women and women with arterial hypertension in the older age group

Chepurnova N.S., Knysh S.V., Rudneva A.V., Zdor V.V., Markelova E.V., Romanjuk P.V., Sayapin S.I.


Multiple studies are currently, aimed at studying the mechanisms of the development of basic age-related pathologies, e.g., arterial hypertension (AH). We found no literature sources concerning associations between the levels of matrix metalloproteinase type 7 (MMP-7), tissue inhibitor of matrix metalloproteinases type (TIMP-1) and anthropometric data which could be useful for diagnosing age-associated, socially dependent diseases and metabolic disorders. The article presents the data on the state of proteolysis system and anthropometric data of 45 almost healthy women and 45 women with arterial hypertension (AH) aged from 59 to 74 years old. The levels of MMP-7 and TIMP-1 in blood serum were studied by sandwich-variant of ELISA test. The waist and neck volume were measured three times with a centimeter tape at an accuracy of 1 cm. Statistical processing of the obtained data was performed using analytical software IBM SPSS Statistics, v. 22.0. The results show some correlations between the levels of MMP-7 and TIMP-1, and the waist and neck sizes (WC). Lower levels of MMP-7 and TIMP-1 were registered in the group of practically healthy women as compared to the second (AH) study group (p < 0.01). Anthropometric data were not markedly diverse: the waist size in the group of healthy women was less than in AH group (p < 0.05), and values of neck volume did not differ significantly between the groups. When assessing the relationships in the group of practically healthy women, we recorded a direct significant correlation between the waist size and neck volume (r = 0.754, p < 0.0001), while in the group of women with AH this relationship was weaker (r = 0.5782, p < 0.0001). In addition, a similar correlation was found between MMP-7 and TIMP-1 levels in the both groups studied. In the group of healthy women this relationship was of medium strength (r = 0.657, p < 0.0001), whereas in the group of women with AH it was stronger (r = 0.720, p < 0.0001). The data obtained suggest the presence of metabolic disorders in the women with arterial hypertension and signs of proteolytic enzyme system disorders.

Russian Journal of Immunology. 2023;26(3):421-426
pages 421-426 views

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