Currently, many human infectious diseases do not developed effective methods of treatment and prevention. One of the latest successes of biotechnology is the use of adenoviral vectors carrying immunodominant antigens  of various pathogens as genetically engineered vaccines  both  preventive and therapeutic. The use of genetic  engineering technologies allows not  to use in the  manufacture of vaccines  live viruses and  bacteria, reduces  the  time  needed for vaccine  creation and  production of new vaccines.  Adenoviral vectors  naturally penetrate into human cells, causing a rather  long and significant  both humoral and cellular immune response. In the second  part of review, we provide  information about  the ongoing  worldwide  clinical  trials of adenoviral vector-based vaccines against various infectious diseases such as influenza, malaria, Ebola haemorrhagic fever, tuberculosis, hepatitis and  several others, like as to consider selection parameters of volunteers, vaccination schedule, doses of drug administration, results of completed experiments, and preliminary data  on currently ongoing  research.

Autoimmune diseases  and  complications  after  transplantation  operations are  major   public health  problem, as they often  lead to deterioration in the quality  of life, reduce  work capacity  and  disability in the  population. To date, the  induction of antigen-specific tolerance is the  promising method of therapy. During this process dendritic cells play important role.  In  this review current data  of tolerogenic dendritic cells characterization, of factors inducing tolerogenic properties in dendritic cells, development of central  and peripheral tolerance by dendritic cells, role of tolerogenic dendritic cells in the formation of regulatory B cells, molecular and cellular mechanisms that are involved in the formation of an immunological tolerance depending on tolerogenic dendritic cells were analyzed. Tolerogenic dendritic cells play a key role in maintaining of immune homeostasis by inducing immunological tolerance mechanisms which  are associated with the  generation of regulatory T cells, induction of anergy or apoptosis of T cells. Formation of tolerogenic properties in dendritic cells is occurred by various factors that regulate the maturation and differentiation of dendritic cells and induce anti-inflammatory agents synthesis of them. Because of their ability to induce natural and peripheral regulatory T cells, dendritic cells contribute to the development of the central  and peripheral tolerance in the organism. Analysis of the results  of experimental work demonstrates that  in addition to induction of regulatory T cells tolerogenic dendritic cells participate in generating of regulatory B cells that  play a role in the  formation of tolerance and  have the  ability  to affect  the  population of regulatory T cells. However, the  mechanisms that lead to the formation of B cell population by tolerogenic dendritic cells have not been completely established yet. Review of published data showed that the molecular and cellular mechanisms of immunological tolerance induction by tolerogenic dendritic cells and other regulatory cell subpopulations interacting with each other are similar to a large extent.  Currently, the role of tolerogenic dendritic cells in maintaining of immune tolerance is determined generally, however, some aspects  require  further  investigation. Subsequent experimental studies will lead to a better understanding of the signaling mechanisms and the realization of immunological functions of tolerogenic dendritic cells, which provide additional information for application approaches development of tolerogenic dendritic cells for the control of autoimmune diseases and transplant complications.

Over last years, a novel strategy for vaccination of people against potentially pandemic influenza A viruses is actively developed worldwide, i.e., a combined (prime-boost) vaccination. It provides  amplification (boosting) of immune response  for a vaccine  be means  of pre-vaccination (priming) with another vaccine.  We have first studied an issue of immunological consequences for people after priming  by live attenuated influenza H5N2 vaccine (LAIV), followed by a boost with inactivated influenza H5N1 vaccine (IIV) 1.5 years later. Unlike non-primed volunteers, the primed persons developed more rapid and high production of serum antibodies (of HAI-, MN-, ELISA-types) after a single vaccination with H5N1 IIV. That concerned induction of antibodies to the H5N1 vaccinal strain A, and other heterologous strains containing H5 haemagglutinin. In primed persons, the antibodies showed  higher  avidity as compared to non-primed individuals. Before inoculation with H5N1 IIV, the  IgG-antibody titers  to A virus (H5N1), and  the  levels of specific  CD4⁺  and  CD8⁺   memory T-cells proved  to be higher  in primed subjects  than  in non-primed persons.  The  boosting  effect  of H5N1 IIV did not correlate with HAI-and MN-based data on immunogenicity of priming  H5N2 live attenuated vaccine.  In general, the results obtained justify a new direction in applications of LAIVs for protection against potentially pandemic influenza virus A.

A key role of T cells in viral elimination and  absence  of strong  T cell responses  in patients with chronic hepatitis C virus (HCV) infection presumes that activation of antigen-specific T cells may be a promising approach to enhance treatment efficacy. Given the central role of dendritic cells (DCs) in the induction of T cell response, the aim of our study was to evaluate  effects of DC immunotherapy upon  immunological parameters in chronic HCV infection. Ten patients with chronic hepatitis C (genotype 1) were vaccinated with monocytederived DCs, generated in presence of IFNα (IFN-DCs) and pulsed with recombinant HCV Core (1–120) and NS3 (1192–1457) proteins. The vaccination protocol included as initiating procedure (one injection per week, ns = 4) and maintaining treatment (one  monthly injection, ns = 6), with subsequent follow up for 6 months. The immunotherapy was not associated with serious adverse events,  significant  post-vaccination reactions, or increased hepatitis C activity, according to biochemical tests. Ex vivo studies have shown that immunotherapy elicited  strong and stable immune response  to Core and moderate response  to NS3 protein, which manifested as a significant  increase  of MNC proliferation and  IFNγ production in response  to Core  and  enhancement of IFNγ production (without higher  proliferation rates),  in response  to NS3.  DC  immunotherapy also led to increase  of ConA-induced MNC proliferation up  to normal levels indicating a recovery  of mitogenic T cell  reactivity. Meanwhile, T cell  activation did  not  elicit  antigen-specific Th2  response  and  expansion of CD4⁺CD25⁺CD127  regulatory T cells.  Despite induced immune response, the  immunotherapy with  DCs was not  accompanied by decreased viremia  levels. Nevertheless, a transitory decrease of viral load  in four patients and stable decrease of viremia  in two patients as well as an inverse correlation between  NS3-specific proliferation and viremia  (Rss = 0.62; p < 0.05) by the end of 6-month follow-up indicated that  the antigenspecific T cells may have a potential to control viral replication.

Natural killer(NK) and  NKT lymphocytes are important components of innate immunity, and compose a first-line defense against cancer. These populations are characterized by high heterogeneity and are divided into several subpopulations, by differences in functional activity as well as CD56  and CD16  expression. Studying  heterogeneity for these  lymphocyte populations in healthy  donors  is important, due  to imbalance between  different  lymphocyte subsets in cancer patients. Changes in the ratio of these subpopulations may be of prognostic and clinical  significance in malignant diseases. The present  study was conducted with peripheral blood  lymphocytes in 50 healthy  donors. When  analysing  population of NK  lymphocytes we have identified 18.0±11.3% of antigen-positive cells, their fluctuations ranged  from 7.6 to 29.2%, whereas average number of cells with  CD3^-CD56⁺ and  CD3^-CD16⁺   phenotypes was equal  to 16,2±8.1%, and  11,0±6.7%, respectively. The  subpopulation analysis  showed  that  the  primary  pool  of NK  cells  was presented by CD56^dimCD16^dim cells  by  52.3±19.9 percent.  We  detected  minor   subpopulations,  e.g.,  CD56^dimCD16^bright,  CD56^-CD16⁺, CD56^brightCD16^- (0.3±0.2%, 1.7±0.9%, and  1.3±0.6%,  respectively). Search  for  intracellular perforin has revealed  that the number of CD56⁺Perf⁺ cells comprized 25.1±14.8%, CD16⁺Perf⁺, 23.8±16.0%. Cytometric analysis showed that perforin is found, predominantly, in CD56^dimCD16^dim NK lymphocytes, whereas the cells with CD56^dimCD16^bright, CD56-CD16⁺, CD56^brightCD16^- immunophenotypes did not produce perforin. For the first time, we have discovered a subpopulation of NK cells with the СD56dimCD16dim immunophenotype that did not contain intracellular perforin (2.0%).  The NKT cell population with СD3⁺CD16/СD56⁺  phenotype was detected in 7.1% (25% – 3.45; 75% – 8.75) antigen-positive cells, within a range of 2.5 to 11.9%. Analysis with a combination of monoclonal antibodies CD3/CD56/CD16 has shown that the number of CD3⁺ CD56⁺ cells was 4.33% (25% – 2.25; 75% – 7.3), whereas the number of CD3⁺CD16⁺ was 3.087% (25% – 0.9; 75% – 6.2). These  data  demonstrate that  the differences in results  between  the CD3/CD16/CD56, and  CD3/CD16 test systems are statistically significant  (p < 0.05). It was shown that the primary-pool NKT-cells are CD56⁺CD16^- cells, whose number is about 5.45% (25% – 2.95; 75% – 7.3) among  total CD3⁺ lymphocyte population. Two minor subpopulations were also detected which differed in expression  of CD56 and CD16 antigens. Hence, the level of CD56^-CD16⁺ cells was 3% (25% – 0.25; 75% – 3.05),  and the number of CD56⁺CD16⁺ was equal to 0.67% (25% – 0.25; 75% – 0.9). Hence, the observed wide phenotypic diversity of NK and NKT-cells reflects their  ability  to exert  various  functional activities.  This  study, showing  high  heterogeneity of NK  and  NKT lymphocytes, may serve as a basis for the study of imbalances between  different  subpopulations of these cells in cancer patients.

Age-dependent development of Th17 and Treg lymphocyte subsets in healthy  humans is studied insufficiently. The  present  study aimed  to investigate  quantitative characteristics of Th17  and  Treg subsets in peripheral blood of healthy  subjects for various age groups.

352 healthy  humans (168 female  and 184 male), one month to 85 years old, were subject to examination, including 79 infants in their first year of life; 34 children at aged 1 year to 2 years 11 months; 24, at 3 to 4 years 11 months; 28, at the age of 5 to 6 years 11 months; 25 children aged 7-8 years 11 month; 36 children aged 9 to 11 years 11 months; 39 adolescents aged 12 to 14 years 11 months; 26 adolescents aged 15 to 18 years; 25 young adults aged 20 to 35 years; 11 adults at 36 to 49 years old; 16 adults aged 59-70 years, and 9 elderly people over 70 years old. The study was performed with capillary blood in children under 2 years, and venous blood taken in elder persons.  The basic and ‘minor’ subsets of peripheral blood lymphocytes were evaluated by flow cytometry using four-color staining of whole blood and following erythrocyte lysis. We used the following surface markers: CD3, CD4, CD8, CD25, CD127, CD161, CD45R0 for lymphocyte subsets detection.

It has been  shown  that  Treg percentage (a ratio  of CD4⁺CD25^hiCD127^low/neg  in the CD3⁺CD4⁺  gate) did not  depend on  age of the  people  under  study, and  can  be approximated by a linear  function. The  absolute number of Tregs in childhood is progressively  decreased and,  after 10 years old, it reaches  plateau  values. This age-dependent relationship may be approximated by a logarithmic function. Evaluation of Th17 subset levels demonstrated a strong  relative  and  absolute  age-dependent growth  of this  cell  subpopulation. Percentage and absolute  numbers of Th17 lymphocyte (share  of CD4⁺CD161⁺CD45R0⁺ in the CD3⁺CD4⁺gate), can be approximated by a square function. The age of 10-12 years seems to be critical to the immune system formation. We suppose  the process of the immune system development to be completed at this age, and maturation of the immune cell populations is then  observed.  A decrease in both relative and absolute  numbers of Treg and Th17 lymphocyte subsets was found in elderly persons (> 70 years old). Our data on peripheral blood Tregs and Th17 subsets, with respect  to their percentage and absolute  numbers in healthy  humans, may be used as age-related reference values.

Granule-mediated cytotoxicity of effector cells is a universal mechanism of tumor growth inhibition and induction of tumor cell death. The aim of present  study was to evaluate  expression  of lytic molecules in DCs generated in presence of IFNα (IFN-DCs), and to analyze  the role of granule-mediated mechanism for IFN-DC cytotoxic activity against tumor cell lines. IFN-DCs were generated by culturing of plastic-adherent peripheral blood mononuclear cells in presence of GM-CSF and IFNα for 4 d followed by LPS addition for 24 h. The tumor cell lines were obtained from malignant tissues from patients with glioblastoma multiforme. Maturation of IFN-DCs in presence of LPS was accompanied by accumulation of intracellular perforin and granzyme B molecules. Perforin expression  showed a direct  correlation with intracellular lysosome-associated membrane  protein-1  (LAMP-1/CD107a)  expression   in  LPS-stimulated  IFN-DCs.   However, CD107a expression  did not increase  under  LPS stimulation. At the same time,  LPS caused  upregulated degranulation in IFN-DCs, as shown  by an increase  of surface  CD107a expression  on IFN-DCs. LPS  activation of DCs generated from the same donors  in the presence of GM-CSF and IL-4  (IL-4-DCs) did not influence perforin and  granzyme B  expression   in  IL-4-DCs which  was  significantly   lower  than  in  IFN-DCs.  Intracellular pool of CD107a molecules was increased in response  to LPS  stimulation of IL-4-DCs, but surface  CD107a expression  did not  change  on IL-4-DCs. Studies  of cytotoxic activity  of LPS-stimulated IFN-DCs revealed that  concanamicyn A (CMA), an  inhibitor of vacuolar  H⁺-ATPase and  of perforin/granzyme B-mediated signaling  pathway, caused  reduced cytotoxicity of donor DCs  towards  glioblastoma cell lines. Involvement of perforin/granzyme B-signaling pathway  into the DCs cytotoxicity was confirmed with glioblastoma cell lines, since blockage  of this mechanism with vacuolar  H⁺ ATPase  blocker (CMA) caused  inhibition of the IFN-DC cytotoxicity. Differently reduced DC cytotoxic activity by CMA may suggest that the glioblastoma cell lysis can be mediated via perforin/granzyme B-independent mechanisms.

Currently actively  discussed  the  role  of innate immunity receptors, in particular TLRs  in the immunopathogenesis of bronchial asthma (BA).

The aim of our work was to study the expression of ТLR2 and TLR4 on the nasal mucosal cells and peripheral blood leukocytes  of patients with BA of different  severity.

The study included 40 children with asthma (3-12 years) and 10 healthy  children. Methods: real-time PCR, flow cytometry and multiplex immunofluorescence analysis evaluated the levels of pro and anti-inflammatory cytokines (IL-1β, IL-1ra, IL-6, IL-8, IL-10, TNFα) in nasal swabs.

The result of the study hyperactivation of the factors of innate immunity at the level of the mucosal  of the nasal cavity in patients with asthma, manifested by increased gene expression  of TLR2, TLR4, and production of proinflammatory as well as anti-inflammatory cytokines. Correlation between cytokine levels and the severity of asthma. In the  peripheral blood  identified a significant  increase  in the  expression  of TLR2  and  TLR4  on circulating CD14⁺  monocytes in children with BA.

Thus,  the  increase  of gene expression  of TLRs  mucosa of the  nasal cavity, increase  surface  expression  of TLR2  and TLR4  on circulating monocytes of patients with bronchial asthma compared to healthy  children. The revealed changes indicate the involvement of the system of TLRs in the immunopathogenesis of bronchial asthma. In the future, TLRs can be used as markers  to predict the course of ad and possible therapeutic targets.

A non-specific stress-induced overload   of  limbic-reticular emotiogenic structures under   the deficiency conditions of endogenous energy  resources is the  main  mechanism for development of asthenic disorders  with  early  developing chronic brain  ischemia among  veterans  of recent  wars,  upon  their  return to civilian  life. We observed  a group  of 30 Afghan  veterans  with  early forms  of chronic brain  ischemia and manifestations of psychogenic asthenic syndrome. When treating psychogenic asthenic syndrome in the veterans of recent  military  conflicts, an adamantane derivative  administered as a course  treatment, at a dose of 50 mg 2 times daily causes psychostimulant, anxiolytic, vegetotropic, antihypotymic, and hypnotic pharmacological effects, with reduction of main psychopathological and somatoneurological symptoms.

A notable immunotropic effect  of the  drug  was revealed, being  expressed  as enhanced immunopoiesis; activation of innate defense  mechanisms; expansion of Treg lymphocyte population, suggestive for increased suppressor  effect  upon  the  mechanisms of autoimmune aggression;  implementation of cell differentiation/ proliferative effect; anti-apoptogenic properties; anti-inflammatory activity (reduction of TNFα, IL-8, hsCRP, along with IL-2  increase) associated with reduced functional activity of mononuclear cells; vasotropic action expressed  as normalized balance of vasoactive  factors  (nitric  oxide,  endothelin-1), decrease of nitrotyrosine concentration, suggesting  the  nitrosylation stress reduction. The  actoprotective drug  effect  is implemented due to a combined psychostimulant, vasotropic, proliferative effects.  It may be expressed  in terms  of higher efficiency and optimization of systemic hemodynamic parameters.

Bronchial asthma is a prevalent chronic allergic disease of lungs at early ages. A priority  task in allergology  is to search  biological  markers  related  to uncontrolled atopic  bronchial asthma. Cytokines fulfill their distinct function in pathogenesis of atopic  bronchial asthma, participating at the initiation, development and persistence of allergic inflammation in airways, causing different  variations of clinical course of the disease (with  respect  to its acuteness, severity, frequency of exacerbations). The  present  work has studied  indices  of cellular  and  humoral links of immunity, as well as levels of some  pro and  anti-inflammatory cytokines in peripheral blood serum (IL-4, IL-10, IL-2 and TNFα), aiming to determine potential markers of uncontrolled atopic bronchial asthma in children. A group of Caucasian (European) children was involved into the research: Cohort 1, moderate atopic  bronchial asthma with controlled course during the last 3 months (n = 59); Cohort 2, severe/moderate-severe atopic bronchial asthma with uncontrolled course of the disease within last 3 months (n = 51),  Cohort 3 – control, practically healthy  children without signs of atopy  (n = 33). All the  children included in the group with atopic  bronchial asthma underwent regular mono/combined basic therapy  at high/ intermediate therapeutic doses.  We performed a comparative analysis  of cell  population indices  reflecting certain cellular  immunity links,  and  determined significantly  lower  levels of CD3⁺   lymphocytes, as well as decrease in relative  and  absolute  contents of CD4⁺  and  CD8+  cells in the  cohort with  uncontrolled course of atopic  bronchial asthma, as compared with controlled-course cohort. When  evaluating concentrations  of cytokines in peripheral blood serum of the patients with controlled and uncontrolled atopic  bronchial asthma, we revealed  significantly  higher  levels of IL-2, IL-4, and  IL-10, as compared to control group.  It was found that TNFα concentration is considerably higher in both cohorts of the patients, being 2to 3-fold higher than the levels of this cytokine in control group.  When comparing the cohorts with different  control of the disease course, we have found that TNFα concentration in the cohort with uncontrolled bronchial asthma is statistically higher  than  among  children with  controllable course  of the  disease.  Hence, the  following  parameters may serve as potential markers  of pediatric atopic  bronchial asthma with uncontrolled course:  low levels of total Т lymphocyte numbers in peripheral blood,  and decreased counts  of CD4⁺, CD8⁺ cells; IgE hyperproduction; low contents of common IgA, and high concentration of TNFα in peripheral blood serum.

Autoimmune mechanisms of Crohn’s disease have been extensively  studied, following discovery of NOD2, ATG16L1, IRGM genetic  polymorphisms associated with  Crohn’s disease.  These  genes play an important role  in  innate immune response  against  intracellular bacteria, in  particular, due  to  their  direct participation in a process  known  as autophagy. Due  to mentioned genetic  traits,  the  CD  patients are more susceptible to chronic infections caused  by intracellular pathogens. Recent studies revealed  high incidence of intracellular infection with Mycobacterium  paratuberculosis and  E. coli in the intestinal tissue specimens and blood  macrophages obtained from  the CD  patients. Such  a chronic, non-resolved infection may disturb  the immune cell properties and  affect  the  balance of pro-inflammatory and  anti-inflammatory cytokines, thus resulting  into chronic inflammation, a hallmark of Crohn disease.

In  this  view, potential  usage  of NK  cells  aimed  for  influencing macrophage activity  represents a new approach in understanding and treatment of autoimmune pathologies. The macrophages are controlled by NK cells. I.e.,  binding  of NKG2D receptor to the MICA  molecules on the macrophage surface causes their lysis.

A signal transfer via NKG2D receptor may increase functional activity of NK against defective macrophages, and hence, promote their elimination. Moreover, in Crohn patients with usually elevated NKG2D⁺  lymphocyte numbers, a stimulation of NKG2D⁺  cells  by soluble  MICA  (sMICA) may  influence the  balance between cytotoxic and regulatory lymphocytes, and reduce  pro-inflammatory cytokine secretion, in order to attenuate chronic inflammation of gut tissues. This review is aimed  to discuss a role of NKG2D⁺  NK  cells in Crohn’s disease pathology and their possible implications for management and treatment of this disorder.