Current issue
3 . 2020
Hot points of immunology

SARS-CoV-2 virus and other epidemic coronaviruses: pathogenetic and genetic factors for the development of infections


At the end of May 2020, more than 6.1 million cases of SARS-CoV-2 virus infection were registered in the world and more than 370 000 were fatal. First time outbreak of new infection occurred among residents of Wuhan, China at the end of 2019. Mortality rate for the current COVID-19 epidemic is significantly lower than for severe acute respiratory syndrome (SARS) or Middle East respiratory syndrome (MERS). However, the SARS-CoV-2 virus spreads much faster than SARS and MERS and causes far more deaths totally than both SARS and MERS combined. Information on factors that affect the development, course and outcome of infections caused by coronaviruses stated in the article was obtained from investigations carried out during the period of previous epidemics of coronavirus infections and during the current SARS-CoV-2 pandemic. The following factors are discussed in the present article: the direct cytopathic effect of viruses, infection of the immune system cells, the role of inflammation in the development of the disease, innate immunity factors, autoimmune reactions, features of the expression of immunoregulatory molecules, the role of host organism factors, including genetic ones.


Combinations of polymorphisms in vascular endothelial growth factor gene and genes of its receptors (VEGF/VEGFR) at development of cardiovascular risk in rheumatoid arthritis patients


Introduction. Cardiovascular risk (CR) development in rheumatoid arthritis patients (RA) is considerably increased. Major factors of CR cannot explain the given phenomenon completely, therefore features of system inflammatory processes which it is characteristic for RA are analyzed. The considered of synovium angiogenesis and of angiogenic signaling pathways, including mediated by vascular endothelial growth factor VEGF and its receptors system VEGFR is one of the important factors of RA immunopathogenesis.

Aim. Analysis of single-nucleotide polymorphisms of VEGF (rs699947, rs3025039), its receptors KDR (rs10020464, rs11133360), NRP-2 (rs849530, rs849563, rs16837641) genes and associations of their combinations with CR development at RA patients.

Material and methods. 135 RA patients (median age is 55 years, duration of disease is 7 years) are surveyed. The increase of CR is revealed at 45.2 % of patients. Genotyping was carried out with restriction analysis of fragment length polymorphism (RFLP) and Real-time PCR (RT-PCR) with TaqMan.

Results. The complex genotypes which frequency are increased in group of high cardiovascular risk patients concerning RA patients with low cardiovascular risk were revealed. The highest relations of chances of high CR development at carriers of complex KDR 14011TT:NRP2 13581TT:VEGF-2578CC, and early relations high CR at patients with KDR 17693CC:KDR 14011TC:NRP2 13581TT:VEGF-2578CA: VEGF+936CC. Protective genotypes, patients with which are steady against high CR development even at long current RA, are revealed also.

Conclusion. Polymorphic positions of angiogenesis mediators genes can in common influence change of certain CR at patients with RA.

Cellular immunology

Generation of murine dendritic cells in vitro using Flt3-L and their characteristics


Introduction. Currently, cell vaccines based on myeloid dendritic cells (mDCs) are being actively developed to modulate an adaptive immune response in order to correct certain human diseases. However, there is still little data on the effectiveness of the use of plasmacytoid dendritic cells (pDCs) for the same purposes.

Aim of the study. Development of the protocol for in vitrogeneration of mice pDCs using recombinant Flt3-L and description of the phenotypic and functional properties of the obtained dendritic cells (DCs) to address the possibility of their use in the development of cell vaccines.

Material and methods. DCs were generated from mouse bone marrow cells using Flt3-L. Frequency and phenotype of the DCs were assessed by flow cytometry. DC’s morphology was assessed using fluorescent microscopy. Treg frequency and CD4+-lymphocytes proliferation induced by DCs were assessed in mixed lymphocyte culture. The expression of CCR9 on DCs was assessed using flow cytometry.

Results. It was shown that the use of Flt3-L provides in vitrogeneration of bone marrow-derived cultures with a relatively high content of B220+CD11c+-pDCs and Sirpα+CD11c+-mDCs. The B220+CD11c+-pDCs and the Sirpα+CD11c+-mDCs obtained differed in morphology and degree of maturity. In a mixed lymphocytes culture, the dendritic cells were able to generate an increased amount of Treg and cause a reduced proliferation of CD4+-lymphocytes, thus manifesting an immature phenotype. Further maturation of the DCs via CpG DNA stimulation led to an increased immunogenicity, according the Treg reduction and enhanced CD4+-lymphocytes proliferation in mixed lymphocytes culture. We found that a relatively small number of B220+CD11c+-pDCs and Sirpα+CD11c+-mDCs carried the CCR9 chemotaxis receptor on their surface, however, the number of B220+CD11c+-pDCs and the Sirpα+CD11c+-mDCs carrying intracellular CCR9 tended to 100 %. Thus, the use of Flt3-L provides the opportunity ofin vitro generation of B220+CD11c+ pDCs and Sirpα+CD11c+-mDCs that can be useful in the field of cell vaccine development to direct an adaptive immune response decreasing undesirable immune reactions or increasing desirable.


Evaluation of cytokines IL-1β and IL-1Ra concentration in nasal secretion in patients with acute purulent rhinosinusitis during immune therapy of β-D-glucans


Introduction. Standard methods and medicines for treatment of acute purulent rhinosinusitis (APRS) do not possess immunomodulating activity that does not promote development of fast dynamics of the end of inflammatory process and knocking over of the basic symptoms of disease. The β-D-glucans mushrooms origin have anti-inflammatory and immunomodulating activity. However, application of these biopolymers for treatment APRS is studied insufficiently.

The aim of the study - an assessment of a distribution of functional pair of cytokines IL-1Ra and IL-1β concentrations in nasal secrets in patients with APRS against application of antibiotics in a combination with β-D-glucans.

Material and methods. Patients have been divided into 4 independent groups. Group 1 consisted of 32 healthy ones. Group 2 consisted of 24 patients with APRS before treatment. Group 3 consisted of 22 patients with APRS who received standard therapy, including antibacterial drugs and puncture of the maxillary sinuses. Group 4 consisted of 24 patients who received β-D-glucans along with standard therapy. The total duration of therapy was 10 days. IL-1Ra and IL-1β concentrations in nasal secrets were defined by enzyme-linked immunosorbent assay, ELISA.

Results. It is positioned that the qualitative visual estimation of shift of a projection of specified cytokines concentrations shows positive changes since by 10 day of treatment in group of the patients received combined therapy, the area of a projection of its concentration comes nearer to cytokines projection area in healthy people. Standard therapy does not lead to such effect, since the minimal impact on local changes of these cytokines levels.

Conclusion. Addition of a medicine containing immunomodulating component β-D-glucans to the APRS standard treatment is reasonable.


Glucosaminylmuramyl dipeptide acid (GMDP-A) modulates intracellular signaling pathways in natural killer cells


Introduction. Medicines developed as derivatives of muramyl peptides have been the subject of experimental and clinical studies for several decades. The intracellular NOD2- receptor responsible for the binding of muramyl dipeptide and its derivatives remains an attractive molecular target for the development of new drugs with a known mechanism of action. GMDP-A belongs to one of the NOD2-receptor ligands. In this study signaling mechanisms initiated by GMDP-A were characterized for the first time in natural killer (NK) cells.

Material and methods. NK cells isolated by magnetic separation from peripheral blood of healthy donors were incubated with 10 pg/ml GMDP-A for 6 hours and 16 hours. The levels of genes expression were determined using the Illumina HumanHT-12v4 Expression BeadChip kit and Illumina iScan instrument (Illumina, USA). The data were statistically processed using an application software package (v 2011.1, Illumina).

Results. It was found that GMDP-A enhanced the transcription of a number of genes associated with transduction of signals from receptors, thereby ensuring the maintenance of the functional activity of NK cells. GMDP-A induced the synthesis of transcripts of the MAPK signaling pathway, VAV2 signal mediators, the CRKL signal adapter, enhanced the synthesis of transcripts of the regulatory subunit 3 (γ) phosphoinositide-3-kinase and phospholipase C B1 genes that control phospholipid metabolism, activated the cells with defects in the cellular immunity link. GMDP-A significantly increases the expression level of the following key genes associated with the control of the mechanisms of direct and antibody-mediated cytotoxic activity of NK cells: CRKL, VAV2, ZAP70, RAP1A, PLCB1, FCGR3A, the MAPK family, IFNA1, TNFRSF9, TNFSF14.

Conclusion. Experimental data justify the high potential of the GMDP-A to counteract altered recognition of the early stages of oncogenic and viral transformation by key cells of the innate immunity, the NK cells.

Clinical immunology

Clinical and immunological effectiveness of an immunotropic drug in children with acute respiratory infections with bronchial obstructive syndrome


Introduction. The incidence of bronchial obstructive syndrome (BOS) against the background of acute respiratory infections (ARI) in young children remains high. This group of children, according to previous studies, revealed a reduced immune response to infection. Thus, it is advisable to use immunomodulating therapy in children with bronchial obstructive syndrome in ARI.

Aim of the study - to evaluate the clinical and immunological effectiveness of the use of the drug alpha-glutamyl-tryptophan spray in children with bronchial obstructive syndrome in acute respiratory infections.

Material and methods. The article presents the results of a randomized, double-blind, placebo-controlled study conducted in the infectious-pulmonological department of the St. Petersburg Children’s Hospital. The study involved children under the age of 6 years with the presence of 2 or more episodes of biofeedback on the background of ARI. Alpha-glutamyl-tryptophan was prescribed after stopping the acute picture of the disease for 5 days in the form of a spray at a dose of 25 pg for children under 3 years old and 50 pg for older than 3 years. Clinical efficacy was assessed by comparing baseline clinical history data with follow-up data 6 months after therapy. The evaluation criteria were the frequency and characteristics of the course of acute respiratory infections and biofeedback, the number of hospitalizations. An immunological study was carried out twice: in the acute period of the disease and not earlier than the 18th day after taking the drug or placebo. The absolute and relative number of lymphocytes and their subpopulations CD3+, CD4+, CD8+ (T-lymphocytes), CD19+ (B-lymphocytes) were determined. Functional activity of polymorphonuclear neutrophils, phagocytic index (AF) and phagocytic number (PS), phagocytosis completion index (PZF) were estimated.

Results. The use of alpha-glutamyl-tryptophan intranasally in the form of a spray in the complex treatment of children with biofeedback led to a reduction in the frequency and duration of acute respiratory infections, helped to reduce the number of episodes of biofeedback against ARI and improved immunological parameters that normalize CD4+-T-cells level, normalize the CD4+/CD8+ ratio, a decrease in the level of pro-inflammatory cytokine TNFα.

Conclusion. Identified positive effects of the drug on the frequency, severity and duration of ARI in children with biofeedback. Favorable changes in immunological parameters allow the use of alpha-glutamyl-tryptophan spray in the treatment of ARI in children with biofeedback and for the prevention of their recurrent course.


Methods for evaluating the specifi c activity of preparations of human antirhesus Rho(D): current status of the problem


Preparations of immunoglobulin human antirhesus Rho(D) are unique means of specific perinatal prevention of Rh-immunization of women with Rh-negative blood affiliation. The pharmacological effect of the drugs is realized by means of antirhesus Rho(D) IgG antibodies (Ab) (anti-D-Ab) that can neutralize Rh-positive fetal red blood cells that have entered the mother’s bloodstream. The choice of a method for evaluating the specific activity (content of anti-D-Ab) in human immunoglobulin preparations plays a key role in ensuring their potential therapeutic effectiveness. Methods of automatic continuous analysis of hemagglutination, enzyme-linked immunoassay and flow cytofluorimetry are used in the world practice to assess the specific activity of human immunoglobulin antirhesus Rho(D), whereas in our country these methods are not implemented in domestic immunobiological drug production. This research analyzes the stages of improvement of methods for evaluating specific activity, features of their reproduction, as well as advantages and disadvantages. Quantitative assessment of the content of anti-D-Ab in International units of activity or micrograms in comparison with the International standard sample of activity of immunoglobulin antirhesus by instrumental method has been established as a necessary condition for obtaining high-quality human immunoglobulin antirhesus Rho(D). The discussion provides the knowledge obtained at the moment, as a result, is justified a conclusion about the prospects for improving the methods for evaluating the specific activity of domestic human immunoglobulin antirhesus Rho(D) preparations in accordance with modern requirements for bioanalytical methods. Scopus, Web of Science, Pubmed, WHO, Global Health, elibrary, EMA, FDA, and the State Pharmacopoeia of the Russian Federation were used to search for literature.


The role of cytokines in the pathogenesis of infective endocarditis


In recent years the interest in studying of the cytokines' immunopathogenetic role in the various infectious and inflammatory diseases formation has not weakened. Our review demonstrates the role of cytokines, produced after a direct contact with the antigen. Cytokines cannot be called as factors that relate only to the immune system. They connect the immune, nervous, endocrine, hematopoietic and other systems to engage in the regulation of a general protective reaction. Cytokines play an important role in hematopoietic, tissue homeostasis, intersystem signal transmission. With the development of inflammation, cytokines demonstrate a high spectrum of biological activity and provide maximum efficiency of those body systems that are required at the moment. Infective endocarditis (IE) is a disease of infectious nature with the primary localization of the pathogen on the heart valves and parietal endocardium. IE occurs with the possible generalization of the septic process and the development of immunopatho-logical manifestations. In this regard, the analyze of current information about the pathogenetic role of cytokines in infective endocarditis is the subject of interest in modern research. For the literature review used the databases: eLIBRARY, CyberLeninka, and Medline (Pubmed).

Short communications

Efficiency of a dense extract of the sum of flavonoids in the form of ointment at treatment contact allergic dermatitis in experiment


The aim of research is to study the specific activity of the preparation of external action of a dense extract of the sum of flavonoids from Bidens tripartitae.

Material and methods. The authors developed and studied a new medical product -a dense extract of the sum of flavonoids in the form of ointment obtained from Bidens tripartita, which was used for treatment of contact allergic dermatitis, experimentally induced in guinea pigs by 2,4-dinitrohlorbenzol.

Results of research have shown that 5 % ointment a medical product on the basis of a dense extract of the sum of flavonoids, visually operated on inflammatory process of integuments of porpoises in comparison of 1 % and 3 % with ointments on the basis of a dense extract of the sum of flavonoids, anthihistamine drug Psilo-Balsam® and glucocorticoid ointments Celestoderm B® century more effectively.

Conclusions. The studied 1, 3 and 5% ointments of dense extract of the sum of flavonoids have a pronounced effect on experimental contact dermatitis in guinea pigs. The greatest therapeutic effect has a 5 % ointment of a dense extract of the sum of flavonoids on a local basis for contact allergic dermatitis. At the same time the index of reducing the severity of skin manifestations (Ind) was the highest compared to other investigated groups - 37.9 %.

Peculiarities of immunopathogenesis of interstitial cystitis/syndrome of a pained bladder


The aim of the study was to determine the expression of cytokines, mast cells quantity in the bladder mucosa and their relationship in patients with IC/BPS.

Material and methods. A total of 126 women with clinically diagnosed IC/BPS were examined, mean age 46.7 ± 14.0 years. The concentration of interleukins (IL)-1β, IL-6, IL-8, tumor necrosis factor a (TNFα) in blood serum was determined by ELISA. Mast cells were identified in biopsy specimens of the bladder mucosa taken during cystoscopy using histological analysis.

Statistical analysis was performed using the Statistica program in Microsoft Excel release 6 (StatSoft, USA). The Pearson correlation coefficient is calculated.

Result. In the main group, moderate pain was observed in 56.3 % of cases, and urination more than 8 times a day in 59.5 % of cases. The average level of pro-inflammatory cytokines -IL-1β, IL-6, IL-8 and TNFα in the blood serum of the examined main group was higher than in the comparison group by 63.34, 66.67, 68.83 and 70.16 % respectively. The number of mast cells averaged 82.27 ± 38.76. Mast cells number correlated with IL-6 and IL-8 weak, with TNFa - a noticeable relationship (r = -0,418). There was a moderate negative correlation between the mast cells number and the level of TNFα (r = -0.418), no significant correlation with the levels of IL-6 and IL-8 was found.

Conclusion. The identification of cytokines in patients with IC/BPS can be used to improve the diagnosis and exclusion criteria for this pathology. Analysis of serum cytokines and mast cells is a convenient approach to monitoring the activation of inflammatory cells in the tissue of the bladder.


Valentina Borisovna Gervazieva (1939–2020)


All articles in our journal are distributed under the Creative Commons Attribution 4.0 International License (CC BY 4.0 license)

Musa R. Khaitov

Corresponding member of Russian Academy of Sciences, MD, Professor, Director of the NRC Institute of Immunology FMBA of Russia