[Vaccinoprophylaxis of chronic somatic diseases exacerbations in groups of risk].

Modem version of I. Mechnikov's hypothesis on association of somatic diseases with infectious agents is presented. List of bacteria and viruses associated with various types of cardiomyopathies, atherosclerosis, gastritis, gastric and duodenal ulcerative disease, type 1 diabetes mellitus. Literature data showing that influenza vaccination reduces number of fatal myocardial infarctions and strokes during winter seasons as well as number of hospitalizations due to exacerbations of chronic cardiovascular and cerebrovascular diseases are summarized. Data on probability of coincidence of influenza vaccination and sudden death in elderly persons are reviewed.

[Prospects for development of immunoprophylaxis up to 2020 - 2030].

Analytic materials on perspectives of immunization up to 2020 - 2030 are presented. Middle-term and long-term prognoses are proposed, perspectives of development of distinct aspects of immunization are considered including increase of number of infections controlled in framework of national immunization schedule, routine immunization of middle-aged and elderly persons, keeping of routine mass vaccination during epidemiological welfare, development and implementation of alternative methods of immunization as well as using new technologies of vaccine manufacturing. It was concluded that to 2020 - 2030 synchronous use of vaccines in national immunization schedules framework will result in elimination of several anthroponoses, decreased incidence of widespread childhood infections down to sporadic cases and significant increase of life-span of patients with chronic diseases.

[Expression of Toll-like receptors in spleen and lymphatic nodes after immunization by mucosal routes].

AIM: To determine level of Toll-like receptors (TLRs) expression in spleen and lymphatic nodes of mice after immunization by mucosal routes. MATERIALS AND METHODS: Mice were immunized with polycomponent vaccine Immunovac either by mucosal or subcutaneous route. Expression of TLRs in spleen, respiratory tract-associated lymphatic nodes as well as in small intestine was measured in immunized mice by flow cytomentry method. RESULTS: After immunization of mice by subcutaneous, intranasal and oral routes level of TLRs expression was different. Significant expression of TLR9 and absence of TLR2 expression was noted after non-parenteral methods of immunization. After oral immunization expression of TLRs was identified in gut-and respiratory tract-associated lymphoid tissue as well as in spleen; after intranasal immunization--in respiratory tract-associated lymphoid tissue, and after subcutaneous immunization--in spleen and respiratory tract-associated lymphoid tissue. CONCLUSION: After oral immunization expression of TLRs was identified in all studied organs, including spleen. Involvement of spleen to this process allows to assume establishment of not only local but also systemic immunity.

[Medical and biological safety assessment of genetically modified maize event MIR604].

There are presented the results of genotoxicologic, immunologic and allergologic examinations which were conducted within the framework of integrated medical and biological assessment of genetically modified rootworm Diabrotica spp.-protected maize event MIR604. Analysis of damages of DNA and structural chromosome aberrations, assessment of the allergenic potential and immunoreactive properties has not confirmed any genotoxic, allergenic and immunotoxic effect of maize event MIR604.

[Acellular pertussis vaccine].

Protective, immunogenic, toxic, and sensitizing properties of acellular pertussis vaccine (aPV) developed according to original technology were studied, aPV had marked protective activity which lasted more than 2 years. Sera of mice immunized by aPV also possess protective properties, and they were more prominent than in sera of mice immunized by pertussis bacteria suspension (PS). Immune sera to aPV neutralized cytopathogenic effect of pertussis toxin (PT) on ovarian Chinese hamster cells in 1:250 dilution, whereas neutralizing activity of sera to PS was very low. Level of antibodies to PT was higher in rabbits immunized, according to schedules and dosage recommended for children, by aPV than by PS. High immunogenicity of aPV was proved also by levels of IgG to PT in sera of mice immunized three times by aPV in human dosage. During experiments on mice and guinea pigs aPV had mild toxicity, did not induce autoimmune process, did not have anaphylactogenic properties compared with bacterial suspension characterized by high anaphylactogenic activity. Histamine-sensitizing abilityof aPVwas 40 times lower than that of PS. Assessment of pyrogenic properties of aPV and PS performed on rabbits showed that aPV was 1,000 times less pyrogenic than PS. Obtained results demonstrate high protective and immunogenic properties of domestic acellular pertussis vaccine and its low toxic and sensitizing characteristics.

[Protective activity of secreted proteins of Streptococcus pneumoniae and Klebsiella pneumoniae].

Protective efficacy of secreted proteins of Streptococcus pneumoniae and Klebsiella pneumoniae cultivated on cardiocerebral broth and semisynthetic growth medium respectively was studied in vivo. Fraction with molecular weight 30 - 50 kDa obtained by the method of membrane fractionation had high protective efficacy. Two-dose immunization of mice with this fraction provided 80 - 100% protection from infection by homologous strains of S. pneumoniae and K. pneumoniae. Cross-protective activity of the fraction was revealed when infecting immunized mice by different K-types of K. pneumoniae. Blood sera of mice immunized with 30 - 50 kDa fraction possessed preventive features protecting from infection 90% of animals while 100% of death in the control group. It was determined that protective efficacy of the mentioned fraction was determined by protein-containing antigens because proteolytic disruption of the protein component resulted in loss of protective properties of the preparation.

[Medical and biological safety assessment of genetically modified maize event MON 88017. Report 2. Genotoxicologic, immunologic and allergologic examinations].

There are presented the results of genotoxicologic, immunologic and allergologic examinations which were conducted within the framework of integrated medical and biological assessment of genetically modified rootworm Diabrotica spp.--protected and glyphosate tolerant maize event MON 88017. Analysis of damages of DNA and structural chromosome aberrations, assessment of the allergenic potential and immunoreactive properties has not confirmed any genotoxic, allergenic and immunotoxic effect of maize event MON 88017.

[Concept of inducing of rapid immunological protection against pathogens].

Concept of construction of new type of vaccines for inducing rapid nonspecific immunological protection against pathogens by activation of innate immunity mechanisms has been formulated. Materials about formula of the concept, theses of theory of innate immunity, and experimental and clinical data, which confirm the concept, are presented. Results of studies with recombinant proinflammatorycytokines and synthetic ligands for Toll-like receptors as well as with polycomponent vaccine containing antigens of opportunistic bacteria (Immunovac VP-4) are also presented. Obtained data allowed to assume that activation of innate immunity mechanisms by preparations carrying pathogen-associated and molecular structures of microorganisms can result in formation of rapid and nonspecific protection against any pathogen.

[Klebsiella pneumoniae secreted protein-containing antigens in the system of adaptive immunity].

The study was aimed at the evaluation of the antigenic properties of K. pneumoniae secreted protein-containing antigens with a molecular weightt of 21 and 34-35 kD, obtained from supernatant culture fluid. As confirmed by the method of flow cytofluorimetry, the protein-containing fractions belonged to the secreted components of the microbial cell. The fraction with a molecular weight of 34-35 kD possessed high antigenic activity and contributed to the formation of specific antibodies after the immunization of mice. At the same time none of the protein fractions lead to an increase in the level of autoantibodies in mouse blood sera to organ-unspecific and organ-specific antigens. As revealed by the method of solid-phase, in 6 (27.3%) from 22 patients of patients with rhizomelic spondylitis had an increased level of IgG to K. pneumoniae cell-wall antigens with a molecular weight of 34-35 kD. An increase in the level of IgG to the secreted protein-containing fraction with a molecular weight of 34-35 kD was detected only in one patient (4.5%) (p<0.05).

[Protective effect of the aerosol immunization of mice with the polycomponent vaccine Immunovac VP-4 after the challenge of the animals with Klebsiella pneumoniae virulent strain].

Used four schemes of the administration of the preparation with different time of the exposition of the animals in an aerosol chamber were tested with their subsequent intraperitoneal challenge with K. pneumoniae virulent strain K16. Irrespective of the number of immunization courses, the administration of the preparation made at intervals of 1 day, or daily, did not ensure any protective effect, but only led to an insignificant increase in their survival time in comparison with nonimmunized animals. After intervals between immunizations were increased to 3 days the protective effect of aerosol immumization was obtained (the survival rate was 65-80 % and considerably differed from that of the controls). The protective effect of aerosol immunization thus obtained was comparable with the effectiveness immunization made in a single subcutaneous injection. Aerosol immunization resulted in low antibody titers to the antigens contained in the vaccine, while after a single subcutaneous injection high antibody titers to Klebsiella and Proteus antigens were detected. The antigen-stimulated blast transformation of spleen lymphocytes in mice subjected to aerosol immunizations in 5 exposures was high. After subcutaneous immunization significant changes in such characteristics were detected on day 15. The data thus obtained were indicative of good prospects in the development Immunovac VP-4 as the medicinal form intended for use in aerosols.

[Immunovac-VP-4, immunomodulator decreases immunosuppression and enhances the cytotoxic activity induced by the cytostatic Cisplatin].

The influence of the vaccine Immunovac-VP-4, prepared from the antigens of opportunistic microorganisms, on the proliferative and cytotoxic activity on peripheral blood mononuclears (PBMN) from healthy donors in vitro and on spleen cells of CBA mice in vivo during their incubation with Cisplatin was studied. VP-4 produced a dose-dependent, stimulating effect on the proliferative potential of PBMN and, when used in the highest of all tested doses (20 microg/ml), increased the Cisplatin-suppressed proliferative activity of PBMN in 9.4-fold. VP-4 increased the cytotoxic activity of PBMN on tumor line cells K-562 (38,4 to 60.1%) and increased the cytotoxic effect of Cisplatin (68.18 to 87.56%). A single injection of VP-4 to mice stimulated the proliferative activity of spleen cells, studied ex vivo, units and partially restored their cytostatic-suppressed activity. The cytotoxic action of the spleen cells of immunized mice on tumor line cells YAC-1 was twice as great as that of spleen cells taken from intact animals and potentiated the cytotoxic action of Cisplatin. The mechanism of increasing the proliferative activity and cytotoxic effect of monomuclears under the influence of vaccine VP-4 is seemingly linked with the synthesis of cytokines, influencing the lymphokine-activated cytotoxicity of lymphocytes.

[Influence of dendrite cells, generated with the use of immunomodulators of microbial origin, on the proliferative and cytotoxic activity of lymphocytes].

The influence of ripe dendrite cells (DC) on the proliferative activity of mononuclear leukocytes and the cytotoxic activity of lymphocytes of syngenic mice was studied. As inducers of DC ripening, the combination of antigenic components incorporated into the vaccine "lmmunovac Bh-4", (Klebsiella pneumoniae, Proteus vulgaris, Escherichia coli, Staphylococcus aureus), as well as K. pneumoniae LPS and TNF-alpha, were used. This study demonstrated that DC activated with these preparations enhanced the proliferative activity of mononuclear leukocytes, the activity of Bh-4 being higher than that of K. pneumoniae LPS. The increase of proliferative activity was accompanied by a rise in the cytotovicity of mouse lymphocytes with respect to the NK-sensitive tumor line YAC-1 or Ehrlich tumor cells. The incubation of the lymphocytes with ripe DC (with the use of the preparations under study as ripening inducers), loaded with tumor antigens, made it possible to obtain cytotoxic lymphocytes having high cytotoxic activity with respect, mainly, to those tumor lines from which lysates for the treatment of DC had been produced. The activation of DC with bacterial immunomodulators led to an increase in the antigen-presenting function of these cells, to their higher capacity for regulating the differentiation of mononuclear leukocytes and for activating the cytotoxicity of natural killers.

[Generation of dendrite cells and the use of immunomodulators of bacterial origin as cell differentiation inducers].

The generation of ripe dendrite cells (DC) of marrow origin was obtained with the use of the vaccine Immunovac-BN-4, an immunomodulator of microbial origin, as well as Klebsiella pneumoniae LPS and TNF-alpha, as ripening inducers. These inducers equally led to the ripening of DC. The generation of ripe DC was characterized by morphological, phenotypical and functional changes. The immunophenotype of cells altered from CD34+, CD38-, CD40-, CD80-, CD86-, MHC I-, MHC II-, F4/80- to CD34-, CD38+, CD40+, CD80+, MHC I+, MHC II+, F4/ 80(low). In parallel with the ripening of DC their phagocytic activity decreased. In culture medium with ripe DC the levels of such cytokines as IL-1b, IL-6, IL-12, IFN-gamma, TNF-alpha significantly increased and the production of IL-4 decreased. The content of IL-2 and IL-10 remained unchanged.

[Immunological characteristics in the therapy of atopic dermatitis in children with polycomponent vaccine Immunovac BN-4].

IgE-mediated reactions linked with the polarization of the immune process towards, mainly, the activation of Th2 lymphocytes which synthesized interleukins, responsible for switching over B lymphocytes to the production of IgE, were found to be the most important mechanism of the pathogenesis of atopic dermatitis (AD). The use of immunomodulating preparations, capable of changing unbalance in the system of Th1/Th2 cells, is one of promising approaches to the complex therapy of AD. Poly-component vaccine Immunovac BN-4 was included into the complex therapy of AD in children, The dynamics of immunological characteristics was studied for the period of 6 months after the end of the course of therapy. A considerable increase in the absolute and relative amount of lymphocytes with markers CD3, CD4, CD16, CD21, CD25, a rise in the levels of IgA, IgG and a decrease in the level of total IgE in the blood serum were established. The inclusion of the polycomponent vaccine into the complex therapy of AD may be recommended.

[Optimization of the therapy of atopic dermatitis by means of immunotherapy].

Taking into account disturbances in the functioning of the immune system in atopic dermatitis (AD) and the potentiating role of staphylococcal and other infections, the possibility of the optimization of the therapy of AD with the use of preparations having immunomodulating action and immunogenic activity is proposed. In the complex therapy of AD in children we used polycomponent vaccine Immunovac B-4, introduced intranasally and orally. Under the influence of immunotherapy the clinical characteristics of the patients had pronounced positive dynamics. A considerable decrease in the spread of the process, the degree of its severity and subjective symptoms was noted shortly after the course of vaccine treatment. Simultaneously the SCORAD index dropped from 64.5 to 39.4. During the later period of observation further decrease in the severity of the course of AD in children occurred, and the minimal characteristics were observed in 6 months of observation. At that time the SCORAD index fell to 19.9 +/- 1.34. The volume of pharmacotherapy and the number of acute respiratory infections considerably decreased, the positive dynamics of the quantitative and qualitative composition of the intestinal microflora was noted. The prolonged immunotherapeutic effect of the polycomponent vaccine made it possible to recommend the vaccine for the optimization of the therapy of AD.

Viruses as nonspecific modulators of immunological reactivity.

As shown experimentally and by examination of patients, numerous DNA and RNA viruses induce transitory nonspecific changes in the immune response of the host to heterologous antigens. Attenuation does not affect this property of the parent strains. Nonspecific immunomodulation is connected with disturbed functions of B and T lymphocytes, macrophages and haemopoietic cells. The actual mechanisms of immunomodulation differ in different infections.

Virus-induced decrease in the activity of serum blocking factor(s): a mechanism of activation of the autoreactive T effectors in graft versus host reaction and of controlling T suppressors.

It has been shown that serum blocking factor(s) (SBF) previously detected in normal mice bind(s) to receptors on the surface membranes of virus-induced autoreactive T lymphocytes (ARTL) and to that of ARTL activity inhibiting T suppressors (TSar). The interaction of SBF with the receptors is reversible, H-2 restricted and associated with the inhibition of functional activity of the tested T lymphocyte populations. In mice during acute tick-borne encephalitis (TBE), in the course of inapparent infections induced by Langat virus or dengue type 2 (D2) and after infection with the attenuated yellow fever virus (strain 17D), the SBF activity significantly decreased, while ARTL and TSar became activated. Administration of normal mouse serum to infected animals with SBF deficiency resulted in inhibition of both inductive and productive phases of ARTL and TSar formations. Based on these findings, the virus-induced decrease in SBF activity may be considered as one of the mechanisms triggering autoimmune responses. The autoreactive pathological states can develop under various endo-and/or exogenic conditions influencing on SBF and on the TSar activity.

Nonspecific T-suppressors in experimental tick-borne encephalitis.

Suppressor cells inhibiting graft-verus-host reaction under conditions of two-way incompatibility were activated in lymphoid organs of tick-borne encephalitis virus-infected mice. Cells with suppressor activity were found in the thymus, peripheral lymph nodes (LN) and spleen but not in the bone marrow, or peritoneal exudate cells adhering to the plastic surface. The cells were identified as T lymphocytes based on the following properties: sensitivity to anti-theta serum, inability to adhere to plastic surface, and resistance to anti-mouse gamma-globulin serum. The T suppressors were activated 3 days after infection (p. i.) in the thymus and LN and at 4 days in the spleen; they were detectable until the appearance of clinical signs of the disease (8-9 days p. i.).

Serum factor blocking the activity of virus-induced autoreactive T lymphocytes of mice.

Normal mouse serum was found to contain a factor blocking in vivo the effect of autoreactive cells accumulating in spleens of mice infected with tick-borne encephalitis (TBE), Langat, dengue type 2 (D2), and attenuated yellow fever (17D strain) viruses. The activity of the factor was manifested only, when the autoreactive lymphocytes had H-2 antigens identical with H-2 antigens of the serum donors. The hypothesis is discussed that serum factor protects the host against clones of autoreactive T lymphocytes generated in viral infections.

Formation of autoimmune effectors after transfer of virus-induced autoreactive precursors to syngeneic intact receptor animals.

We verified the assumption that autoimmune responses underlie immunosuppression developing in the course of virus infection. It has been shown by adoptive transfer that administration of Langat-virus-induced autoreactive lymphocytes (ARL) to the syngeneic recipient is followed by accumulation of autoreactive effectors (AE) eliciting a graft-verus-host reaction (GVHR) in the syngeneic system (AEgvhr). In addition, cytotoxic T-lymphocytes (AEc) appeared against the syngeneic macrophages. This effect was mediated by the H-2 restriction principle for ARL donors and their recipients. The infection triggered a T-suppressor-mediated reaction inhibiting the differentiation of precursors into mature autoreactive effectors. In the absence of T-cell suppressor effect (on consecutive syngeneic transfer) ARL will cause systemic pathologic changes in the recipients, namely, infiltrative and necrotic lesions in different organs and tissues. This mechanism is attributed to the activation of precursors in the body of intact recipient and to the formation of mature effectors owing to cytokines released by donor ARL.