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Background: The problem of identifying vaccine-specific T-cell responses is still a matter of debate. Currently, there are no universal, clearly defined, agreed upon criteria for assessing the effectiveness of vaccinations and their immunogenicity for the cellular component of immunity, even for healthy people. But for patients with inborn errors of immunity (IEI), especially those with antibody deficiencies, evaluating cellular immunity holds significant importance., Aim: To examine the effect of one and two doses of inactivated adjuvanted subunit influenza vaccines on the expression of endosomal Toll-like receptors (TLRs) on the immune cells and the primary lymphocyte subpopulations in patients with common variable immunodeficiency (CVID)., Materials and Methods: During 2018-2019, six CVID patients received one dose of a quadrivalent adjuvanted influenza vaccine; in 2019-2020, nine patients were vaccinated with two doses of a trivalent inactivated influenza vaccine. The proportion of key lymphocyte subpopulations and expression levels of TLRs were analyzed using flow cytometry with monoclonal antibodies., Results: No statistically significant alterations in the absolute values of the main lymphocyte subpopulations were observed in CVID patients before or after vaccination with the different immunization protocols. However, after vaccination, a higher expression of TLR3 and TLR9 in granulocytes, monocytes, and lymphocytes was found in those patients who received two vaccine doses rather than one single dose., Conclusion: This study marks the first instance of using a simultaneous two-dose vaccination, which is associated with an elevated level of TLR expression in the immune cells. Administration of the adjuvanted vaccines in CVID patients appears promising. Further research into their impact on innate immunity and the development of more effective vaccination regimens is warranted.

Background: for the first time, the effect of one and two doses of adjuvanted influenza vaccines on toll-like receptors (TLRs) in patients with common variable immunodeficiency (CVID) was studied and compared (primary vaccination with one vs. two doses, primary vs. repeated vaccination)., Materials and Methods: Six patients received one dose of quadrivalent adjuvanted influenza vaccine during the 2018-2019 and 2019-2020 influenza seasons, and nine patients with CVID received two doses of trivalent inactivated influenza vaccine during 2019-2020. Expression of TLRs was measured by flow cytometry., Results: The expression of toll-like receptors in patients with CVID was noted both with repeated (annual) administration of the influenza vaccine and in most cases was accompanied by an increase in the proportion of granulocytes (TLR3 and TLR9), lymphocytes (TLR3 and TLR8), and monocytes (TLR3 and TLR9). When carried out for the first time as a simultaneous vaccination with two doses it was accompanied by an increase in the proportion of granulocytes, lymphocytes expressing TLR9, and on monocytes-TLR3 and TLR9., Conclusion: in CVID patients, the use of adjuvanted vaccines is promising, and research on the influence of the innate immunity and more effective regimens should be continued.

Aim: Study subpopulation structure of lymphocytes in healthy individuals under the effect of various influenza vaccines in an in vitro system., Materials and Methods: Evaluation of immune- phenotype features of PBMCs, activated in vitro by immune-adjuvanted and unadjuvanted vaccines against influenza in healthy individuals, was carried out by using flow cytometry method., Results: Grippol plus vaccine caused a more pronounced stimulating effect compared with subunit and split-vaccines on NK-cells, cells with markers of early activation CD45/CD25, induced the quantity of natural regulatory cells (CD4/CD25/Foxp3), increase of the number of B-cells and reduced the amount of cell types with apoptosis marker CD45/CD95., Conclusion: Immune-adjuvanted vaccine Grippol plus induced formation of effectors of both innate and adaptive immunity and possessed the most powerful potential regarding activation of various types of immune-competent cells compared with unadjuvanted vaccines.

Aim: Study the effect of Vaxigrip split, Influvac subunit and Grippol plus immune-adjuvanted vaccines on the content of myeloid (mDC) and plasmacytoid (pDC) dendritic cells (DC) in blood of vaccinated healthy women. Materials andmethods. Blood of 30 healthy women aged 18-50 years was studied at days 7 and 30 after the vaccination. pDC (CD14+CD16-/CD85k(ILT3)-PE/ CD123-PC5) and mDC (CD14+CD-16-/CD85k(ILT3)-PE/CD33 -PC5) immune phenotyping was carried out using mAbs (Beckman Coulter, France) and flow cytometer Cytomix FC-500 (Beckman Coulter, USA)., Results: Use of unadjuvanted vaccines Vaxigrip and Influvac resulted in an increase of the numbers of mDC and pDC (p<0.05) in blood of the vaccinated only at day 7 of the observation. Grippol resulted in a more significant (2.2 - 3.6 times, p<0.05) increase of DC subpopulations (compared with unadjuvanted vaccines) at both day 7 and a month after the vaccination., Conclusion: Influenza vaccination activated innate effectors - the first component on the way of infection penetration - dendritic cells of both myeloid and lymphoid origin. Wherein, a more pronounced and prolonged effect of such activation is observed when immune-adjuvanted vaccine is used compared with subunit and split vaccines.

Aim: Study innate and adaptive immunity in patients with migrating erythema, clinical effectiveness ofcombined therapy using Immunovac vaccine and dynamics of immunologic parame- ters as a result of the therapy., Materials and Methods: 37 adult patients with migrating erythema were examined. The patients were divided into 2 groups: 1st gr. (14 individuals) - Immunovac by intranasal-subcutaneous method against the background of basic therapy; 2nd gr. (23 individuals) - 200 mg/day doxycycline therapy for 21 days. Phagocytic activity of blood neutrophils; TLRs expression on mononuclear leukocytes of peripheral blood (PBML) and skin cells in foci by flow cytometry with mAT against TLR2, 3, 4, 5, 6, 7, 8, 9 using flow cytometer FC-500; subpopulation composition of peripheral blood lymphocytes; levels of pro-, anti-inflammation and regulatory cytokines in blood sera by EIA method; IgG, 1gM, IgA in blood sera were studied in patients before treatment and 1 month after therapy., Results: A high level of TLR2, 4, 7, 8 on skin cells in foci, TLR2, 4 - on blood cells; low content of CD95+ and CD25+, high-level of serum IL-lb, IL-2 and IL-4, an increase of general IgE level was detected in patients'with migrating erythema. Immunovac facilitated an increase of CD95+ and CD25+, IFN-y synthesis, reduced the level of general IgE in a more pronounced way than basic therapy., Conclusion: Inclusion of Immunovac into therapy of patients With migrating erythema facilitates increase of clinical effectiveness and correlates with correction of immunologic disorders.

The article reports survey data on 23 patients with erythemas, including 19 patients with herpes-associated erythema multiforme (HAEM) and 4 patients with Darier's erythema annulare centrifugum (DEAC). Patients in the initial state (baseline) and after two weeks of therapy with proteflazid were characterized by measuring the levels of Toll-like receptor (TLR) expression in peripheral blood mononuclear cells (PBMC) and in epithelial cells of the throat and the skin. The TLR expression in PBMC and skin was assessed by flow cytometry with monoclonal antibodies (ICA) (Caltag Laboratories, USA; Hycult Biotech, Netherlands) against relevant antigens. In addition, patients were also characterized by the content of subpopulations of lymphocytes expressing surface markers CD3, CD4, CD8, CD16, CD21, CD23, CD72, CD25, and HLA-DR in the peripheral blood, which was measured by flow cytometry. The therapy with proteflazid in patients with both HAEM and DEAC led to normalization of the level of both T-cell and B-cell immunity, which was manifested by an increase in the total number of lymphocytes, CD3+, CD4+, CD21+, and CD72+. Measurements of the dynamics of TLR expression in the course of immunotherapy showed an increase in the number of TLR 2, 3, 4, 7, 8, and 9 in PBMC (which was especially pronounced for TLR2) and in epithelium of the pharyngeal mucosa and skin (increased expression of TLR3, 7, and 9).

Aim: Study of the interrelation between the presence of immune deficiency and development of complications during vaccination of newborns with BCG vaccine., Materials and Methods: In 24 children with complications of vaccine process in the form of cold abscess and lymphadenitis indicators of lymphocyte subpopulation levels were studied by flow cytofluorimetry on Beckman Coulter cytofluoriemter by using monoclonal antibodies with markers CD45+CD3+ - T-cell, CD45+CD3+CD4+ - T-helpers, CD45+CD3+CD8+ - T-supressors-cytotoxic killers, CD45+CD3 CD16+CD56+ - natural killers, CD45+CD3-CD19+ - B-lymphocytes. The level of IgG, IgA, IgM in sera was determined by immune diffusion method in agar by Mancini., Results: In 4 children selective deficiency of IgA, in 5 - hyper-IgM syndrome was detected, which is an innate immunodeficiency and is characterized by the lack of sera IgA, reduction of IgG level and increase of IgM. In 9 children a reduction of CD16+ natural killer lymphocytes was detected, in some cases combined with a reduction of CD8+ T-supressors-cytotoxic killers., Conclusion: The reason of development of complications during BCG administration is the presence of immunodeficiency in children. In these children severe course of the vaccine process, presence of axillary lymphadenitis was observed, therapy of these children continued from 4 to 6 months.

Aim: Detection of features of functioning ofinnate and adaptive immunity pathways in patients with Darier erythema annulare centrifugum (EAC)., Materials and Methods: 14 EAC patients aged 14 - 52 years were examined. The patients were ranked based on therapy variant. The first group consisted of 6 patients who had received Immunovac-VP-4 (Immunovac) against the background of basic therapy; the second group (4 patients) received cagocel against the background of basic therapy; the third group (4 patients) received only basic therapy; the group of healthy individuals consisted of 15 individuals. All the patients had the level ofcytokines in blood sera determined by solid-phase EIA by using Biosource (Austria) test-systems. Evaluation of TLR expression in peripheral blood mononuclear lymphocytes and keratinocytes was carried out by flow cytometry method by using monoclonal antibodies (Catlag Laboratories, USA) against the corresponding antigens; evaluation of content of lymphocyte subpopulations in blood was carried out by using monoclonal antibodies by flow cytometry method on FacsCalibur flow cytometer (Becton Dickinson, USA)., Results: Immunotherapy by Immunovac and cagocel facilitated the increase of CD3+, CD4+, CD8+. Immunovac facilitated a significant increase ofinitially low values ofCD25+, CD95+ and normalization of CD72+; normalization of IgM level. Immunovac increased the level of serum IL-2, induced the increase of IFN-gamma synthesis in contrast to cagocel and basic therapy, the administration of those caused its decrease. TGF-beta increased during the course of Immunovac immunotherapy and decreased during basic therapy. In the course of basic therapy a significant increase of initially high level ofcytokine IL-I beta was observed., Conclusion: Immunovac therapy resulted in correction of content of lymphocyte populations, sera cytokines, facilitating the normalization of immunocompetent cell proliferation processes, activation of NK-cells, macrophages and at the same time suppression of DTH reactions. Immunovac facilitated the enhancement ofTLR3, 9 expression in the skin that indicates the inclusion ofintracellular receptor mechanisms of innate immunity.

Aim: Determine levels of sIgA, IgG, IgA in vaginal secretion and saliva of women of reproductive age with chronic inflammatory diseases of small pelvis organs (IDSPO) at exacerbation stage and remission period., Materials and Methods: Clinical-laboratory and gynecological examination of 105 women was carried out. Based on the results obtained 3 groups were formed: patients with IDSPO at exacerbation stage; patients at remission stage; clinically healthy women. sIgA, IgG, IgA parameters were studied in vaginal secretion and saliva in women with IDSPO at exacerbation stage and remission period by radial immune diffusion in gel by Manchini method., Results: An increase of immunoglobulin level in vaginal secretion of women with IDSPO at remission period and a sharper increase of these parameters during exacerbation of the disease compared with women of the control group were detected. During analysis of sIgA, IgG, IgA levels in saliva in the same groups of women the results were obtained that give evidence that the presence of IDSPO and local immune reaction do not lead to the changes of these parameters., Conclusion: The obtained parameters on the dependence of an increase of immunoglobulin levels in vaginal secretions and the degree of intensity of the inflammatory process give basis to use them with the aim of additional diagnostics.

Aim: Determine subpopulation composition of blood lymphocytes and the level of expression of TLR2 and TLR9 by epithelial cells of cervical canal mucous membrane in women of reproductive age with inflammatory disease of small pelvis organs (IDSPO) at exacerbation stage and remission period., Materials and Methods: Clinical-laboratory and gynecological examination of 105 women was carried out and 3 groups were formed based on the results: patients at IDSPO exacerbation stage; patients at remission stage; clinically healthy women. By using real time PCR, TLR2, TLR9 gene expression levels were determined in epithelial cells of cervical canal mucous membrane in women of all the 3 groups. Subpopulation composition of blood lymphocytes was determined by flow cytofluorimetry by using monoclonal antibodies with CD45+ CD3+ -T-cell, CD45+ CD3+ CD4+ -T-helper, CD45+, CD3+, CD8+ -T-suppressors-cytotoxic killers, CD45+, CD3-, CD16+, CD56+ natural killers, CD45+, CD3-, CD19+ -B-lymphocytes. Immune fluorescence reaction evaluation was carried out in flow cytofluorimeter Cytomics FC 500 (Becton Coulter, USA)., Results: The level of expression of TLR2 gene in the studied groups of patients was established not to differ significantly from parameters in the comparison groups, however it should be noted that this parameter in women with IDSPO at exacerbation stage (causative agents of the infectious process--ureaplasma, staphylococcus, candida) was somewhat higher than in the comparison group. Significantly high level of TLR9 gene expression in cervical canal epithelial cells was detected to correlate with the presence of infectious causative agents. In the group of women with exacerbation of the infectious process the expression of TLR9 was 14.5 times higher compared with the group of women without IDSPO. Among groups of women with IDSPO significant differences in relation to control group in relative and absolute levels of CD3+ T-lymphocytes; CD4+ T-helpers; CD8+ cytotoxic killer T-suppressors, B-lymphocytes compared with the same parameters in clinically healthy women were not detected., Conclusion: The increase of TLR9 gene expression level in cervical canal cells of women with IDSPO may serve as an additional diagnostic feature of the presence and degree of severity of the disease.

Aim: Assessment of results of immunization against measles and mumps in children with recurrent respiratory infections., Materials and Methods: Levels of IgG against measles and mumps viruses were measured using enzyme immunoassay. Two hundred and twelve serum samples obtained from 6 groups of children with 20 - 45 persons (boys and girls) in each were tested. Children of various ages were presented in each group. Also, immunologic parameters were measured in all children., Results: It was established that mean antibody titers to measles and mumps viruses did not change during 10 years. In more than 50% of children correlation between high and low titers of antibodies of different specificity was found., Conclusion: Recurrent respiratory illnesses determine complex regulation of transition of memory cells into cells secreting antibodies to measles and mumps viruses.

Markers of humoral and cellular immunity in 16 patients with vaccine-associated paralytic poliomyelitis (VAPP) were evaluated. Signs of immunodeficiency (decrease of T- and B-lymphocytes counts, impaired synthesis of immunoglobulins, defects of phagocytosis, decrease of NK number) were revealed in all of the patients. Majority of them (81.3%) had defects in humoral immunity. Decrease of CD31, CD4+ and CD8+ was detected in 86.7, 35.7 and 91.7% of the patients respectively. Study of serum immunoglobulins performed in 15 patients showed decrease of IgG, IgM and IgA levels in 6 (40%), 1 (6.7%) and 6 (40%) of the patients respectively. Agammaglobulinemia was diagnosed in one patient in which only trace quantities of IgA and IgG were detected and IgM level was well below the normal. Congenital deficiency of IgA was diagnosed in 3 children. Majority of the children (11 from 12) had comorbidities (frequent respiratory infections, dermatitis, changes of intestinal microflora). Thus, immunocompromised condition of a child is a risk factor for VAPP after administration of alive oral poliovaccine.

32 patients with atopic dermatitis (AD), complicated by pyoderma, were treated with purified staphylococcal toxoid (PST). Changes in clinical and laboratory characteristics in the course of treatment were evaluated. PST was shown to produce a satisfactory therapeutic effect, arresting the symptoms of local staphylococcal infection. An increase in the levels of alpha- and gamma-interferons and decreased content of CD25+ lymphocytes were found. Thus prospects appear for using this preparation as an interferon inductor, as well a for the immunotherapy of AD patients sensitized to Staphylococcus aureus.

The dynamics of immunological characteristics after the administration of polycomponent vaccine B[symbol: see text]-4 to elderly persons, constituting a risk group with respect to acute respiratory diseases and exacerbations of chronic inflammatory diseases of respiratory organs, was studied. The nasal-oral administration of the vaccine induced immunological shifts in the systems of local and systemic immunity. The content of the populations of lymphocytes with markers CD3, CD4, CD16, CD20 was found to have positive dynamics. Considerable shifts in the system of local immunity were registered: the content of sIgA and IgA in the saliva greatly increased; in addition, an increase in the titers of antibodies to Staphylococcus aureus, Klebsiella pneumoniae and Escherichia coli was observed in persons with initially low titers.

The content of IgA, IgM, IgG and the level of specific antibodies in the blood serum of patients with bronchopulmonary diseases after a course of immunotherapy with polycomponent vaccine B[symbol: see text]-4 was studied. A rise in the concentration of IgM due to the synthesis of specific antibodies to Proteus vulgaris, Escherichia coli and Klebsiella pneumoniae was found to occur. The examination of sick children revealed that a high proportion of them (54%) showed a pronounced decrease in the level of IgA. The use of the preparation made it possible to enhance the level of IgA.

The systemic and mucosal immune status in 50 patients with all-year-round allergic rhinitis (AYRAR), chronic infectious rhinitis (CIR) and non-atopic eosinophilic rhinitis (NER) was evaluated. Patients with concomitant pathology (bronchicial asthma, chronic bronchitis, sinusitis, etc.) were found to have changes in the systemic immunity. Patients with AYRAR and CIR had a high level of SIgA in their nasal secretions, and in the latter--also an elevated level of local IgG NER patients exhibited a considerable decrease in local production of immunoglobulins. In the nasal secretions of AYRAR and CIR patients high phagocytic activity of neutrophilis was established in the nitro blue tetrazolum test. The results of the study showed the expediency of determination of immune disturbances in these patients with a view to administration of immunocorrective therapy.