Chronic pelvic inflammatory disease (PID) in women remains a problem due to the importance of medical consequences. The study of the receptor apparatus of neutrophilic granulocytes (NG) involved in anti-infective protection in diseases of various etiologies seems to be relevant. Aim: to clarify the features of variants of quantitative and phenotypic changes in subsets of NG CD11b+CD64-CD32+CD16+, CD11b+CD64+CD32+CD16+ of immunocompromised women during exacerbation of chronic PID of various etiologies. We were tested women 20-40 years: Study Group 1 (SG1, n = 20) – chronic PID during the exacerbation with mono- or mixed latent/recurrent various viral infections (chronic herpes-virus infections, papillomavirus infection, recurrent ARVI); Study Group 2 (SG2, n = 30) – chronic PID of bacterial etiologies; Comparison Group (CG)– 20 healthy women. The number of subsets CD11b+CD64-CD32+CD16+NG (major) and CD11b+CD64+CD32+CD16+NG (minor), receptor expression density (MFI) was determined (FC500, USA). It was found that in PID during the period of exacerbation, diagnostically significant differences in the subset composition of NG were revealed. We got a decrease in the CD11b+СD64-СD32+СD16+NG subset and in 7,6 times increase in the CD11b+CD64+CD32+CD16+NG subset in SG2 with chronic PID of bacterial etiology, in contrast to chronic PID occurring in combination with recurrent/persistent viral infection SG1. Negative transformation of NG subsets is associated with a predominant decrease in the level of expression of the activation CD16. The absence of an adequate response to the infectious and inflammatory process was revealed – the absence of an increase in the expression of the activation CD11b in the major subset in SG1, as well as in the minor subset in groups SG1 and SG2. In the major subset of NG in groups SG2 a decrease in the expression of the activation marker CD11b. In the various viral infections and PID (SG1), in the negatively altered minor subset of NG we got a decrease of expression of CD16, an increase of expression of CD64 and CD32. Determination of subsets of CD11b+СD64-СD32+СD16+, CD11b+CD64+CD32+CD16+NG and their phenotype can be used as diagnostic markers for the differential diagnosis of PID of viral and bacterial etiology, and for the development of new methods of targeted immunomodulatory therapy.

Failure of anti-infectious immune protection is considered a reason for the prolonged course and recurrence of chronic infectious and inflammatory diseases of pelvic organs (PID). Our aim was to evaluate the effect of an original hexapeptide (HP) on negatively altered subpopulations of neutrophil granulocytes (NG) CD11b+CD64-CD32+CD16+ and CD11b+CD64+CD32+CD16+, their phenotype and associated effector functions in immunocompromised women with PID. 35 women (20-40 years old) with PID were studied during the period of clinical exacerbation (study group 1, SG1). Study group 1a (SG1a) consisted of patients who underwent treatment including the HP injections (45 mcg/ mL, 1 ml intramuscularly once a day for 10 days). The comparison group (CG) consisted of 20 conditionally healthy women. The numbers of CD11b+CD64-CD32+CD16+NG and CD11b+CD64+CD32+CD16+NG cell subsets and the density of receptor expression, phagocytic and microbicidal function of NG were determined. In SG1, decreased counts of the major NG subpopulation (CD11b+CD64-CD32+CD16+NG) was revealed (p 0.05), with a trend for increase of minor subset CD11b+CD64+CD32+CD16+NG (p 0.05). In the CD11b+CD64-CD32+CD16+NG subset, we noted a decreased expression of CD16 (1.4-fold), CD11b (2-fold) (p1, 2 0.05). In the minor subset CD11b+CD64+CD32+CD16+NG, the expression densities were decreased in CD16 (1.7-fold), CD11b (2.1-fold, p1, 2 0.05). At the same time, the phagocytic and microbicidal functions of NG were found to be decreased. In the course of immunomodulatory therapy with the HP-based drug, positive changes in immunological parameters were revealed. In SG1a, an increased number of major NC subset was observed, with an increase in the expression density of CD16 by 1.2 times, CD11b by 1.7 times relative to SG1 (p1, 2 0.05). The contents of minor NG subset tended to decrease, along with CD16 expression density reaching the indices of comparison group. CD11b increased 1.3 times relative to SG1 (p 0.05). Higher ratios of actively phagocytizing NG and their killing ability have been registered. Clinically, we observed faster regression of clinical PID exacerbation symptoms and decreased frequency of relapses 6 months after treatment in 88.6% of cases. The positive immunomodulatory effects of the HP-based drug upon altered subsets of CD11b+CD64-CD32+CD16+ and CD11b+CD64+CD32+CD16+ NGs, their phenotype and associated effector functions suggest an opportunity of its usage for the correction of NG dysfunctions in immunocompromised women with PID, thus providing stable clinical and immunological remission and protective effect.

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.

The key role of neutrophilic granulocytes (NG) in the pathogenesis of COVID-19 makes them new targets for therapeutic approaches and of influencing the course and outcome of the disease, restoring changes in the phenotype and functions of NG. Synthetic peptides or polypeptide complexes of action are the most promising in the treatment of COVID-19. Aim: to reveal the effects of the influence of the hexapeptide (HP) – Arginyl-alpha-Aspartyl-Lysyl-Valyl-Tyrosyl-Arginine on the phenotype of functionally significant NG subsets in moderate COVID-19.The study examined patients 61 (57-71) years old (n = 45) in the acute period of COVID-19 – study group1 (SG1). In vitro, samples SG1 were incubated with HP (106 g/L, 60 min, 37 °C) – study group2 (SG2). The number of NG subsets was evaluated: CD16+IFNα/βR1+CD119+, CD16+IFNα/βR1+CD119- , CD16+IFNα/βR1+CD119+, CD64- CD16+CD32+CD11b+, CD64+CD16+CD32+CD11b+ and phenotype by membrane receptor expression density (MFI) (FC 500, Beckman Coulter, USA); NG phagocytic activity was tested before and after incubation with HP. The comparison group (GS) – of 22 volunteers examined in the pre-COVID period.It was revealed that unidirectional effects of HP in vitro contributing to the restoration of the phenotype of subsets CD16+IFNα/βR1- CD119+, CD16+IFNα/βR1+CD119- to CG indicators. There was a decrease in MFI CD16 (p < 0.05) in both subsets; MFI CD119 (p < 0.05) in the CD16+IFNα/βR1- CD119+NG subset, MFI IFNa/βR1 in the CD16+IFNα/βR1+CD119- NG subset. The effects of HP on the phenotype of CD16+IFNα/βR1+CD119+NG subsets in 76% of cases were manifested by a decrease in MFI CD16 (p<0.05), an increase in MFI IFNα/βR1 and CD119 (p1, 2<0.05), and in 24% of cases a decrease in MFI IFNα/βR1 (p<0.05). HP in vitro remodeling of the phenotypes subsets CD64- CD16+CD32+CD11b+ and CD64+CD16+CD32+CD11b+ were established, providing the usefulness of effector functions from hyperactivated to normal. In the CD64- CD16+CD32+CD11b+ subset, there was a decrease in MFI CD16 and CD11b to the indicators CG (p1, 2 < 0.05). Recovery of the NG phenotype under the influence of HP led to the restoration of the phagocytic function of NG.Positive effects of HP in vitro on the phenotypes of subsets actively and NGfunctions in COVID-19 open up prospects for the creation of new methods of immunotherapy to restore NG dysfunctions.

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 IFN/R, IFNR(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+IFN/R1-CD119+, CD16+IFN/R1+CD119-, CD16+IFN/R1+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+IFN/R1-CD119+, CD16+IFN/R1+CD119-, CD16+IFN/R1+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+IFN/R1+CD119+NG subset and phenotype transformation of CD16+IFN/R1-CD119+NG, CD16+IFN/R1+CD119-NG subsets. Increased density expression of CD16, IFN/R1, 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 recIFN2b 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.

Acute destructive pneumonia (ADP) is a severe purulent and septic infectious disorder of childhood, characterized by a high level of morbidity and associated with imbalance of the immune system (IS). Hence, there is an obvious need to study the immunopathogenesis of this disease in order to develop new therapeutic strategy aimed at eliminating the pathogen, detoxifying the body, relieving respiratory failure and correcting functional immune deficiency. Our aim was to perform a clinical and immunological study in order to evaluate efficiency of immunomodulatory therapy using a medical drug with hexapeptide as an active substance. This drug was included into the complex postoperative treatment of children with acute destructive pneumonia. Clinical and immunological examination of 15 children 2-5 years old with ADP was performed before (study group 1 – SG1) and after (study group 1a – SG1a) combined postsurgical treatment including immunomodulatory therapy with a Hexapeptide-based pharmaceutical (HP, Arginyl-alpha-Aspartyl-Lysyl-Valyl-Tyrosyl-Arginine). Comparison group (CG) included twenty healthy children. The contents of T and B lymphocytes, natural killer cells (NK) were measured by means of flow cytometry (CYTOMICS FC 500, USA). Serum levels of IgA, IgM, IgG (ELISA), phagocytic and microbicidal activity of neutrophil granulocytes (NG) were also evaluated. Prior to the treatment in SG1 patients, a decreased number of CD3+CD19-T lymphocytes, CD3+CD8+ TCTL lymphocytes was revealed along with significant decrease in the contents of CD3-CD16+CD56+ NK (p1-3 0.05). It was found that in children with ADP, the IgG level did not differ from indices of control group, with a decrease of IgM (p1, 2 0.05), and increased level of IgA (p 0.05). We have also found a deficiency of NG effector functions, i.e., insufficiency of active phagocytic NG with impaired capture and killing of bacterial antigen. Assays of NADPH-oxidase activity showed lacking response to both inflammation and additional induction by S. aureus. After complex treatment including immunomodulatory therapy in the SG1a group, we revealed a recovery in CD3+CD19-T lymphocyte contents, CD3+CD8+TCTL lymphocytes, CD3-CD16+CD56+ NK (p1-3 0.05). The trends towards normalization of IgA, IgM, improved NG effector functions (killing ability) were revealed due to activation of NADPH-oxidases. The restoration of immunological parameters in ADP was associated with earlier recovery from the purulent-destructive process in lungs, absence of postoperative complications including the prevention of septic process. The clinical and immunological effects of the immunomodulatory therapy program with HP-based pharmaceutical preparations suggest its potential usage during postoperative period in immunocompromised children with ADP.

Post-COVID syndrome (PCS) is a multisystem inflammatory condition with manifestations of chronic fatigue syndrome (CFS) and cognitive disorders (CD), along with reactivation of chronic herpesvirus infections (HVI). The PCS manifestations require studying the molecular mechanisms associated with the production of IFN and receptor functions of neutrophil granulocytes (NG), which is relevant and promotes the search for immunotherapeutic strategies in patients with PCS. Our objective was to study the in vitro effects of recombinant interferon α2b (recIFNα2b) on the phenotype of CD16+IFNα/βR1-CD119+, CD16+IFNα/ βR1+CD119+ subsets and functional activity of NG in patients with post-COVID syndrome and herpesvirus infections. Materials and methods: 45 patients (24-60 years old) with PCS and HVI (HSV 1, EBV, HHV6, CMV) comprised the study group 1 (SG1). A questionnaire was conducted to assess the severity of PCS symptoms using a point scale. We performed a study of the content and phenotype of NG subsets, i.e., the CD16+IFNα/βR1-CD119+, CD16+IFNα/βR1+CD119-, CD16+IFNα/βR1+CD119+ subpopulation, phagocytic and NADPH oxidase function of NG before and after in vitro incubation with recIFNα2b (50 IU/ µL, for 60 min, at 37 °C) in the study group 1a (SG1a). The comparison group (CG) of 30 volunteers examined during the pre-COVID period. Results: We revealed more pronounced clinical manifestations of CFS and CD in SG1 patients with mixed HVI, than in mono-HVI cases. Increased expression density of all receptors was registered on CD16+IFNα/βR1+CD119-NG and CD16+IFNα/βR1-CD119+ NG, thus suggesting the NG activation with initiation of cytotoxicity or NETosis, a decrease in phagocytic function and intensity of NADPH oxidase activity with depletion of NG reserve capacity in SG1. We have obtained some data on the positive effect of recIFNα2b in vitro (SG1a), e.g., decreased CD16 expression density and enhancement of IFNα/βR1 receptor expression in the CD16+IFNα/βR1+CD119- subset. In the CD16+IFNα/βR1-CD119+ subset, we have found persistence of increased MFI CD16 and MFI CD119 receptors, restoration of defective NG phagocytic function and reduced excessive activity of NADPH oxidases. Conclusion: The positive effects of the recIFNα2b influence on deficient function of NG in PCS patients suggest an oppoptunity of using immunotherapy with a recIFNα2b-based drug, combined with highly active antioxidants for treatment of various PCS manifestations including CFS, CD, HVI, thus, probably, ensuring adequate functioning of antiviral and regulatory mechanisms of the immune system.

Acute peritonitis (AP) is among the most frequent and severe conditions in pediatric abdominal surgery. Due to development of antibiotic resistance and increasing number of atypical infectious and inflammatory diseases (IIDs), a lot of specialists suggest combined treatments for these patients which should include not only surgical and etiotropic approaches, as well as therapy aimed at correction of functional defects of immunity. Neutrophilic granulocytes (NGs) reepresent a unique population of cells of primary anti-infectious immune response. Functional NG defects in pediatric AP play a leading role in development, prevalence, severity of peritoneal inflammation, and response to the therapy. Special role is given to functionally significant NG subsets responsible for triggering and implementation of phagocytosis and microbicidal properties of NG in purulent lesions and inflammatory process in children. There is an urgent need for development of new approaches to targeted immunomodulatory therapy in order to correct the NG dysfunction. The aim of the present study was to arrange the programs of immunomodulatory therapy after surgical treatment of immunocompromised children with various forms of acute peritonitis followed by subsequent evaluation of its clinical and immunological efficacy. The study included 12 immunocompromised children aged 5-12 years with different clinical course of acute peritonitis. The study group 1 included patients with local nonrestricted AP; study group 2 involved children with diffuse AP. The comparison groups consisted of 6 children who received standard therapy, i.e., clinical comparison groups 1 and 2, matched for sex, age and diagnosis. A control group consisted of 18 conditionally healthy children at similar age. Clinical examination included collection of the patient’s history, complaints, objective examination and clinical course assessment of the underlying disease. Immunological study included determination of receptor, phagocytic and microbicidal activity of NCs; assessment of NC subpopulations by their numbers and phenotype using flow cytometry, i.e., the cells co-expressing CD64, CD16, CD32, CD11b, with testing density of these membrane receptors by the MFI approach. Targeted immunomodulatory therapy programs were applied for treatment of children with unrestricted local and diffuse AP, taking into account clinical features of AP, as well as changes in number and phenotype of NC subpopulations, and impairment of their effector function. The standards of postsurgical treatment in the children with various forms of AP included different courses of treatment with Imunofan (Hexapeptide – arginyl-alpha-aspartyl-lysyl-valyl-tyrosyl-arginine; HP) using different schedules and duration. We have shown high clinical and immunological efficiency of these therapeutic programs. Thus, reversal of adequate NG functioning was observed, including positive rearrangements of negatively transformed functional NG subpopulations. In this respect, a positive clinical effect was noted in children with atypical AP with various clinical courses, i.e., absence of postsurgical complications, rapid regression of intoxication signs, normalization of body temperature, reduced volume of antibiotic therapy and shorter hospitalization terms.

Investigation of molecular mechanisms associated with interferon (IFN) production and receptor function of neutrophil granulocytes (NGs) in COVID-19 is highly relevant because it can be promising in the search for new therapeutic strategies targeting NGs and their reactivity to restore and strengthen the innate immune response against SARS-CoV-2. Objective. To assess the effects of recombinant IFN-α2b on the phenotype of CD16+IFNα/βR1–CD119+, CD16+IFNα/βR1+CD119–, and CD16+IFNα/βR1+CD119+ NGs from peripheral blood of patients with COVID-19 in an in vitro experiment. Patients and methods. We analyzed blood samples from 31 patients with a mean age of 61 years (range: 57;71 years) with moderate COVID-19. We assessed the number of CD16+IFNα/βR1–CD119+, CD16+IFNα/βR1+CD119–, and CD16+IFNα/βR1+CD119+ NGs, receptor density (FC 500, ‘Beckman Coulter,’ USA), phagocytic activity of NGs before and after incubation with recombinant IFN-α2b. We also measured serum levels of several cytokines, including IFNα, IFNγ, IL-6, IL-8 (ELISA, ‘Vektor-Best’ LLC). The control group comprised 22 adult healthy individuals with a mean age of 58 years (range: 57; 70 years). Results. Patients with moderate COVID-19 demonstrated low serum levels of IFNα and IFNγ along with elevated levels of IL-6 and IL-8. We observed transformation of 3 phenotypes among NG subpopulations: CD16+IFNα/βR1–CD119+, CD16+IFNα/βR1+CD119-, and CD16+IFNα/βR1+CD119+. We observed positive remodulating effects of recombinant IFN-α2b on the number and phenotype of NG subpopulations and their phagocytic activity in our in vitro experiment. Conclusion. Recombinant IFN-α2b demonstrated positive effects in in vitro experiments; therefore, it can be considered in the future as a potential therapeutic tool for moderate COVID-19. Restoration of type I IFN might be an effective treatment option for COVID-19, because it can promote faster virus elimination, restore normal functioning of the IFN system, and have positive regulatory effects on the phenotype of NG subpopulations. Key words: neutrophil granulocytes, subpopulations, COVID-19, phenotype, recombinant interferon α2b

Numerous studies over last decade have shown that functional capacity of neutrophil granulocytes (NG) determines the course and outcome of many diseases. Identification of phenotypic variants of functionally significant NG subpopulations is a new approach allows us to assess the adequacy or deficiency of NG involvement into infectious inflammation processes at molecular level. An opportunity of reorienting a deficient NG subpopulational phenotype in purulent inflammatory diseases due to the rearrangement of the receptor set induced by various immunotropic substances may serve as a key to recovery of normal NG functioning.Our aim was to study the effect of glucosaminylmuramyldipeptide (GMDP) under in vitro conditions upon the phenotypic profile of four functionally significant subpopulations, i.e., CD62L+CD63-NG, CD62L+CD63+NG and CD64-CD32+CD16+CD11b+NG, CD64+CD32+CD16+CD11b+NG, along with assessment of expression density of appropriate membrane molecules and NG microbicidal activity in the children with purulent inflammatory diseases. 90 samples of peripheral blood (PC) were taken from children 2 to 4 years old, including 12 children with minor purulent infection (MPI), and 7 children were studied as conditionally healthy controls. Their peripheral blood was incubated for 60 minutes at 37 °C with GMP (10-6 g/l). Using flow cytometry technique, the relative numbers of some NG subpopulations, i.e., CD64-CD16+CD32+CD11b+NG, CD64+CD16+CD32+CD11b+NG, CD62L+CD63-NG, CD62L+CD63+NG were evaluated, and the phenotype features of each subpopulation were investigated according to the density of appropriate membrane molecule expression (MFI). In parallel, phagocytic and microbicidal activity of NG was tested in these study groups. The obtained data indicate for presence of for distinct NG subpopulations, both in healthy children and in children with MPI. We have revealed phenotypic transformation of the four studied NG subpopulations from MPI patients including disturbed phagocytic and microbicidal functions of the cells. Using of this in vitro system, we have shown that the transformed phenotype of the four functionally significant NG subpopulations of MPI patients was re-arranged under GMDP treatment. At the same time, the number of CD62L+CD63+NG and CD64-CD32+CD16+CD11b+NG subpopulations was increased, along with decreased amounts of CD64+CD32+CD16+CD11b+NG and CD62L+CD63-NG subpopulations, being accompanied by restoration of microbicidal activity of NGs.The obtained data allow us to accomplish current understanding of immunotropic effects of GMDP, and to extend the potential scope of its experimental and clinical application. The new data on GMDP effects revealed by in vitro system, i.e. phenotype rearrangement of functionally significant NG subpopulations CD64-CD16+CD32+CD11b+, CD64+CD16+CD32+CD11b+, CD62L+CD63-, CD62L+CD63+ in atypical purulent inflammatory diseases in children, may be used in the future in order to develop innovative strategies of immunotherapy aiming for correction of NG dysfunction in children with MPI.

Children with clinical signs of immunocompromise have impaired functioning of the immune system. At the same time, the state of subpopulations of multifunctional neutrophil granulocytes (NG), which provide antiviral and antibacterial protection, in such patients has been poorly studied. Objective of the study: to assess the features of the transformation of the phenotype of the three most significant subpopulations of NG, their association with impaired phagocytic and microbicidal activity in immunocompromised children with atypical viral-viral co-infections and bacterial infections, and to clarify their differential diagnostic significance. Materials and methods of research: the phenotypes of NG subpopulations expressing CD64, CD32, CD16, CD11b receptors, phagocytic and microbicidal activity of NG were studied in the peripheral blood of 43 immunocompromised children of both sexes aged of 5–9 years old suffering from recurrent ARVI, atypical chronic mono – or mixed herpes virus infections (HVI) and purulent bacterial infection. 3 study groups were formed: 1st – repeated ARVI/HVI mono, 2nd – repeated ARVI/HVI mixed, 3rd – minor purulent infection (MPI) and a comparison group (20 apparently healthy children). Results: different differentiated transformation of the phenotype of functionally significant subpopulations of NG associated with impaired functional activity of NG as well as with the incidence of viral co-infections (repeated ARVI and recurrent HVI mixed) and the severity of clinical manifestations of MPI has been identified in the studied groups. Conclusion. Еvaluation of the features of transformation of phenotype of NG subpopulations, their effector functions in immunocompromised children with atypical viral-viral co-infections and purulent bacterial infections, will allow in the future to optimize the methods of target immunotherapy aimed at remodulating the negatively transformed phenotype of NG subpopulations, correcting their dysfunctions and, thereby, significantly increase the clinical effectiveness of therapeutic and preventive measures.

In the conditions of steadily increasing urbanization and emerging negative microclimates, there is increase the morbidity of children in megacities. The damaging effect of environmental factors causes the development of “minimal persistent inflammation” of mucosa. We had studied 55 children in age 1–8 years suffering from recurrent ARVI associated with different chronic HVI (HSV I/II, EBV, CMV, HHV VI) and chronic recurrent bacterial diseases. The comparison group was 40 conditionally healthy children, comparable by sex and age. Clinical and immunological methods: ELISA, PCR, cytofluorimetry were used. We studied 2 groups of children: 1 group – 1–4 years old (early children) and 2 group – 5–8 years old, suffering from recurrent ARVI of a high rate of frequency. Clinical and anamnestic data showed that the incidence of ARVI was in 1 group – 7–15 and in 2 groups – 7–24 or more episodes per year, and the duration of these episodes was: 7.5 and 15.8 days, respectively. In all children, repeated ARI in combination with chronic respiratory diseases with frequent exacerbations were associated with recurrent and/or mono- and mixed HVI. Comparative analysis showed that in the children from group 1 demonstrate deficient in IFN-α, serum IgA and various NG disorders. However more severe disorders in IFN-α, NK-cells, NG system were detected in group 2, which is associated with a greater age of infectious history. Our study revealed the urgency to create new approaches for targeted immunoprophylaxis for immunocompromised children living in the megapolis.

Defects in the functioning of neutrophilic granulocytes (NG) are accompanied by various purulent-inflammatory diseases that have a persistent-relapsing course. The bactericidal activity of NG, the processes of intracellular and extracellular killing of bacteria, is directly dependent on their phenotypic features: the expression level of functionally significant membrane receptors CD64, CD16, CD32, CD11b triggering effector processes. It was shown that there are subsets of NG with different phenotypic characteristics and the course and outcome of infectious and inflammatory diseases directly depends on their usefulness. Despite the large amount of accumulated information, the question of the transformation of NG subpopulations phenotype during NG response formation in children with local purulent processes of soft tissues - a small purulent infection (SPI) remains poorly understood. Purpose: to evaluate the local and systemic effector function of neutrophilic granulocytes (phagocytosis, NET formation) with clarification of the phenotype of functionally significant CD64-CD16+CD32+CD11b+, CD64+CD16+CD32+CD11b+ and CD64-CD16+CD32-CD11b+ NG subpopulations in peripheral blood (PB) in children with SPI. Materials and methods. We evaluated the phagocytic function of NG in 60 PB samples of children 5-9 years old, on the 2-3rd day of acute purulent process, before surgery, in smears - prints taken from the local focus of purulent inflammation and 17 PB samples of conditionally healthy children. Also the ability to form NET was additionally determined in smears-prints in children with SPI. In parallel, in PB we evaluated:% NG simultaneously carrying CD64, CD16, CD32, CD11b receptors and their expression density (MFI) by flow cytometry (CYTOMICS FC500, Beckman Coulter, USA). Statistical data processing was performed in the Microsoft Exel 2016 and Stat Plus 2010 software packages. Nonparametric criteria were used: median (Me) and interquartile range (Q1; Q3), Mann-Whitney U-test and Wilcoxon test. Results . We found a significant increase in the content of the NG subpopulation with a negatively transformed phenotype CD64-CD16+CD32-CD11b+ with defects in the expression of CD64 and CD32 receptors, which is the cause of inadequate inclusion of NG in inflammatory processes, disruption of phagocytic activity at the systemic and local levels and the occurrence of SPI in children. The formation of NET in the first stage is necessary to supplement the effector function of NG in order to compensate for disorders of phagocytosis.

Relevance. Neutrophilic granulocytes (NG) are the first cells of the immune system to migrate to the tumor and are actively involved in the implementation of a full-fledged antitumor response through the mechanisms of direct killing of tumor cells, extracellular lysis (NET), and through the activation of antibody-dependent cellular cytotoxicity (ADCC), inhibition of angiogenesis, initiation of other cells with antitumor activity. The aim of the study was to study the effect of cytokines IFN, IFN, G-CSF on the CD62L+CD63- and CD62L+CD63+ subsets and the microbicidal activity of NGs in patients with colorectal cancer (CRC) in vitro. Materials and methods. We studied samples of peripheral blood (PB) of 10 patients of both sexes 38-70 years old with newly diagnosed untreated CRC stage II-III (study group) and 10 healthy volunteers (comparison group). The subsets CD62L+CD63+ NG, CD62L+CD63- NG were assessed by flow cytometry (CYTOMICS FC500, Beckman Coulter, USA), the microbicidal functions of NG were tested by cytochemical methods: activity of NADPH - oxidases, myeloperoxidase (MP), level of cationic protein (CP) in spontaneous tests and under additional stress of S. aureus . The effect of IFN, IFN, G-CSF cytokines on subsets and the microbicidal activity of NG in vitro was studied in both study groups. Microsoft Exel 2016 and StatPlus 2010 were used for statistical processing of the obtained data using nonparametric tests: Me (Q1; Q3), Mann-Whitney U-test and Wilcoxon test . Results . The features of transformation of CD62L+ CD63-NG and CD62L+ CD63+ NG subsets of PB in CRC have been established, that allows to get an idea of the NG ability to roll and readiness to activate the microbicidal arsenal, various defects of spontaneous and induced microbicidal activity of oxygendependent and oxygen-independent mechanisms of NG. The effects of cytokine influence on NG in CRC in vitro have been shown, which indicates the possibility of regulating the receptor and microbicidal functions of NG, and, on the other hand, suggests defects in NG perception of regulatory stimuli, that is confirmed by the progression of tumor growth.

Antimicrobial activity of neutrophilic granulocytes (NG) is based on effective recognition and elimination of microbial pathogens, as well as on complex intracellular signal transduction pathways interconnecting these processes. NG dysfunction leads to emergence of atypical infectious and inflammatory diseases recalcitrant to standard interventions, which requires new vector platforms aimed at restoring normal NG functioning and overcoming antibiotic resistance. Moreover, we emphasize about special interest paid to the NOD1 and NOD2 agonist polymuramyl and NOD2 agonist glucosaminylmuramyldipeptide. Objective of the study was to compare effects triggered by NOD1 and NOD2 agonist polymuramyl and NOD2 agonist glucosaminylmuramyldipeptide on phenotype of neutrophilic granulocyte subsets CD64- CD16+CD32+CD11b+, CD16+CD62L+CD63-, CD16+CD62L+CD63+ in the in vitro system. 64 samples of peripheral blood (PC) collected from 8 apparently healthy children (4 boys and 4 girls) aged 3 to 9 years were examined by flow cytometry (FC 500, Beckman Coulter, USA) assessing NG surface receptors CD64, CD16, CD32, CD11b, CD62L, CD63 with MonAb (Beckman Coulter International S. A., France) by analyzing NG number (%) expressing receptors examined, density of receptor expression measured as mean fluorescence intensity (MFI). For this, there were assessed intact peripheral blood NG from apparently healthy children (comparison group) as well as those exposed to polymuramyl (PM) (at concentration of 10-6 g/l) or glucosaminylmuramyl-dipeptide (GMDP) (at concentration of 10-6 g/l) for 60 minutes at 37 °С temperature. Comparative analysis of surface receptor expression was performed on CD64-CD16+CD32+CD11b+, CD16+CD62L+CD63- and CD16+CD62L+CD63+ NG subsets that suggested about positive transformation of activation parameters in circulating NG exposed to NOD1 and NOD2 agonist polymuramyl as well as NOD2 agonist glucosaminylmuramyldipeptide. At the same time, similar effects of varying intensity were revealed manifested as increased count of NG subsets CD16+CD62L+CD63+ bearing increased level of CD16 and reduced CD62L expression, as well as differences uncovered as significantly increased expression of surface membrane molecules CD16 and CD11b in CD64-CD16+CD32+CD11b+ NG subset from apparently healthy children exposed to polymuramyl as well as increased surface CD32 expression after incubation with GMDP.

Disruption of neutrophilic granulocyte (NG) functioning underlies one of the key causes for negative polymicrobial synergism during virus-microbial co-infections. In connection with this, it is important to develop experimental models for viral-bacterial co-infections in vitro, which might allow to uncover NG involvement in effector events and assess reorganization of inter-connected functionally relevant NG receptors in response to various immunotropic agents. Understanding molecular mechanisms related to any molecule including drug molecules provides its safe use allowing them to become drug of choice. NG functional activity is associated with surface receptors CD64, CD32, CD16, CD11b, which are assigned to several NG subsets exhibiting distinct phenotypes, and their activation leads to complex processes of pathogen elimination. Study objective: to elucidate types of phenotype transition for NG subsets CD64-CD32+CD16+CD11b+ and СD64+CD32+CD16+CD11b+ and assess an opportunity for phenotype reprogramming exposed to hexapeptide arginyl-alpha-aspartyl-lysyl-valyl-tyrosyl-arginine (НP) in experimental in vitro model of viral-bacterial coinfection. Materials and methods. We examined 39 samples of peripheral blood (PВ) collected from healthy adult volunteers (7 women, 6 men) aged 21 to 32 years, subdivided into 3 groups: comparison group 1 (intact NG); comparison group 2 – model of viral-bacterial infection; Study group – to assess HP effects. Model of viral-bacterial co-infection was created by incubating PВ samples sequentially with dsRNA (10-7M) for 60 min followed by fMLP (10-7M) for 60 min, 37 °С. To assess HP effect, PC samples preincubated with dsRNA and fMLP were next exposed to HP (10-6 g/L) for 60 min at 37 °С. We analyzed percentage of CD64-CD32+CD16+CD11b+ and СD64+CD32+CD16+CD11b+NG subsets as well as receptor expression density (MFI) by flow cytometry (FC 500, Beckman Coulter, USA) using MAbs (Beckman Coulter International SA, France). Results. Transformation of CD64-CD32+CD16+CD11b+ and СD64+CD32+CD16+CD11b+NG subset phenotype was experimentally revealed in virus-bacterial model manifested as upregulated expression of all receptors examined. Our data on HP effects ambiguously demonstrated phenotype transformation in CD64-CD32+CD16+CD11b+NG, СD64+CD32+CD16+CD11b+NG in in vitro viral – bacterial coinfection model contributing to recovery of proper NG response.

В учебно-методическом пособии дана характеристика врожденных и приобретенных интерферонопатий, приведена классификация интерферонопатий с учетом, как дефицита интерферонов различного типа, так и гиперпродукции интерферонов I типа, описаны особенности клинических проявлений различных видов интерферонопатий, рассмотрены дифференцированные подходы, как к таргетной терапии интерферонопатий I типа, ассоциированных с гиперпродукецией интерферона альфа, так и интерферонкорректирующей терапии. Учебно-методическое пособие «Врожденные и приобретенные интерферонопатии» составлено на основании Федерального государственного образовательного стандарта высшего образования по специальности 31.08.26 «Аллергология и иммунология (уровень подготовки кадров высшей квалификации)» (МЗ РФ, 2014), написано на современном уровне, с использованием результатов современным научных исследований по данной проблеме и может быть рекомендовано для студентов, клинических ординаторов и аспирантов медицинских вузов, слушателей факультетов непрерывного медицинского образования, врачей аллергологов-иммунологов, акушеров-гинекологов, педиатров, терапевтов, отоларингологов, инфекционистов, врачей общей практики, врачей клинической лабораторной диагностики