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1.
Artigo em Russo | MEDLINE | ID: mdl-30695390

RESUMO

AIM: Evaluation ofthe ability of capsule polysaccharides (CP) of Streptococcus pneumoniae serotype 3 and 14 and their synthetic structure analogues, conjugated with bovine serum albumin (BSA), to detect antibodies in post-vaccination sera of mice. Materials andmethods. Oligosaccharides correspond- ing to one, one and a half and two repeating links of serotype 3 and 14 S. pneumoniae CP were synthe- sized, their conjugates with BSA were produced by squarate method as well. Ligand content-per BSA molecule was controlled by MALDI-TOF spectrometry. Immune sera were obtained after 2 intraperi- toneal administrations to mice of glucoconjugates adsorbed on aluminum hydroxide or 13-valent pneumococcal conjugated vaccine. Determination of levels of post-vaccination class G antibodies and their sub-isotypes was carried out in EIA. RESULTS: Immunization of mice with neoglucoconjugates resulted in formation of predominantly IgGl recognizing serotype 3 and 14 S. pneumoniae C. IgG1 in mice immunized with a 13-valent conjugated vaccine recognized serotype 3 S. pneumoniae CP, but detected serotype 14 S. pneumoniae CP weakly. All the conjugated synthetic oligosaccharides were characterized by a high ability to bind antibodies in blood of mice immunized with the polysaccharide conjugated vaccine. BSA-tetrasaccharide of serotype 3 S. pneumoniae and BSA-tetrasaccharide of serotype 14 S.pneumoniae were characterized by the highest ability to detect IgG1 against C. CONCLUSION: Synthetic oligosaccharides, conjugated with BSA protein-carrier, may be used to develop diagnostic test-systems for determination of antibodies in post-vaccination sera.


Assuntos
Anticorpos Antibacterianos/imunologia , Cápsulas Bacterianas/imunologia , Imunoglobulina G/imunologia , Vacinas Pneumocócicas/imunologia , Polissacarídeos Bacterianos/imunologia , Streptococcus pneumoniae/imunologia , Vacinação , Animais , Camundongos , Camundongos Endogâmicos BALB C , Vacinas Pneumocócicas/farmacologia , Polissacarídeos Bacterianos/farmacologia
2.
Artigo em Russo | MEDLINE | ID: mdl-26259269

RESUMO

AIM: Study epitopic specificity of synthetic disaccharide, recurring link of serotype 3 S. pneumoniae, conjugated with bovine serum albumin (BSA). MATERIALS AND METHODS: Conjugate of the synthetic disaccharide with BSA was obtained by squarate method. Antigenic activity of the conjugate was studied in competitive EIA. Titers of IgG against capsule polysaccharide of serotype 3 S. pneumoniae were determined in EIA by using sera of mice immunized twice with disaccharide conjugate sorbed onto aluminum hydroxide. RESULTS: Disaccharide conjugate used as a well-covering antigen (4 µg/well) in EIA was characterized by a high degree of specificity and interacted only with IgG against serotype 3 S. pneumoniae in antimicrobial sera of animals without reacting with antibodies (ABs) against other pneumococcus serotypes (6B, 10A, 19A, 19F, 23F). Disaccharide conjugated with BSA was determined in competitive EIA to inhibit bonding of ABs to disaccharide by 78.8%, bacterial capsule polysaccharide by 56.9%, BSA did not inhibit the sera activity. The study of sera of mice immunized by serotype 3 S. pneumoniae disaccharide conjugate in EIA, where capsule polysaccharide was used as a plate-sorbed antigen, has established the presence of IgG against capsule polysaccharide at a titer of 1:1600. CONCLUSION: The disaccharide that is a single recurring link of serotype 3 S. pneumoniae contains a key epitope of capsule polysaccharide. The synthetic disaccharide could be used as a component of multivalent conjugated pneumococcal vaccines and for development of diagnostic test-systems.


Assuntos
Dissacarídeos/imunologia , Infecções Pneumocócicas/imunologia , Streptococcus pneumoniae/imunologia , Vacinas Conjugadas/imunologia , Animais , Anticorpos Antibacterianos/imunologia , Cápsulas Bacterianas/imunologia , Bovinos , Dissacarídeos/síntese química , Epitopos/imunologia , Humanos , Camundongos , Infecções Pneumocócicas/prevenção & controle , Sensibilidade e Especificidade , Sorogrupo , Soroalbumina Bovina/administração & dosagem , Soroalbumina Bovina/imunologia , Streptococcus pneumoniae/patogenicidade , Vacinação , Vacinas Conjugadas/administração & dosagem
3.
Artigo em Russo | MEDLINE | ID: mdl-25536778

RESUMO

AIM: Study experimental production series of Staphylovac-2 by accumulation of specific IgG and safety. MATERIALS AND METHODS: Experimental production samples of staphylococci vaccines were studied by the accumulation of specific IgG in sera of immunized BALB/c line mice in EIA. Safety was evaluated in tests of acute and chronic toxicity including pathomorphologic and histologic, hematologic and biochemical studies, studies of the effect on central nervous system. RESULTS: A statistically significant (2.6 - 3.0 times) increase of IgG levels in sera of immunized mice compared with control was noted. In the experiments studying acute and chronic toxicity the increase in body mass and mass of internal organs differed from data obtained from control animals at no observation periods. None of the studied methods of safety evaluation showed differences of the studied vaccine series from the control. CONCLUSION: The recommended dose for subcutaneous administration into human of 200 µg is experimentally justified and could be the basis for carrying out clinical studies of staphylococci vaccines in humans.


Assuntos
Imunoglobulina G/sangue , Vacinas Antiestafilocócicas/administração & dosagem , Vacinas Antiestafilocócicas/imunologia , Animais , Formação de Anticorpos , Humanos , Imunização , Camundongos , Vacinas Antiestafilocócicas/efeitos adversos
4.
Bull Exp Biol Med ; 157(5): 612-5, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25257424

RESUMO

We studied the effects of immunization with a conjugate of carrier protein and hexasaccharide ligand related to a fragment of capsular of Str. pneumoniae serotype 14 polysaccharide chain on activation of innate and adaptive immunity. It was found that two-fold immunization with the glycoconjugate adsorbed on aluminum hydroxide significantly increased the titer of IgG antibodies to capsular polysaccharide in the blood and protected 100% mice from infection with Str. pneumoniae serotype 14. Enhanced bactericidal activity of peripheral blood lymphocytes of mice was found 4 and 24 h after the first immunization with the immobilized glycoconjugate. Adsorption of the glycoconjugate on aluminum hydroxide resulted in modification of the immune processes at the stage of activation of innate immunity and subsequent strengthening of the adaptive immunity.


Assuntos
Glicoconjugados/farmacologia , Polissacarídeos/química , Streptococcus pneumoniae/química , Animais , Sequência de Carboidratos , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular
5.
Artigo em Russo | MEDLINE | ID: mdl-25816515

RESUMO

AIM: Study the effect of aluminium hydroxide on molecular-cell mechanisms of innate immunity activation and its adjuvant effect on immunogenicity of natural bacterial and synthetic pneumococci antigens. MATERIALS AND METHODS: Surface markers of dendritic cells (DC), mononuclear leukocytes (ML) and cytokine levels were determined by flow cytometry; IgG titers--by EIA. Protective activity was evaluated in experiments with active protection of mice from infection with virulent pneumococci strains. RESULTS: Aluminium hydroxide increased the ML content of mice spleen expressing TLR2 and TLR4. Its addition into the culture of immature DC induced the appearance of a population of cells with mature DC markers--CD83, CD80, CD86, however, the level of undifferentiated cells (CD34) and cells with adhesion molecules (CD11c, CD38) did not change. DC produced IL-1ß, IL-5, IL-10, IFNγ into the cultivation medium. An increase of cytokine production took place 2 hours after the administration into mice and was retained for the observation period (24 hours). Th1 (IFNγ, TNFα) and Th2 (IL-5, IL-10, GM-CSF) cytokine production gave evidence on immune response polarization by Th1/Th2, type. After 2 administrations of aluminium hydroxide into mice the number of ML with CD19+, CD5+, NK1.1+, CD25+, MHCII+ markers increased during decrease of CD3+, CD4+ and CD8+ T-lymphocytes. Adaptive immunity activation was characterized by high IgG titers to pneumococci capsule polysaccharide and protection of 90 - 100% of the mice against infection with lethal doses of S. pneumoniae strains, was detected during 2-fold immunization of mice with conjugates of synthetic pneumococci oligosaccharides with BSA,sorbed onto aluminium hydroxide, whereas natural bacterial antigens provided 90 - 100% survival of animals during immunization without the adjuvant. CONCLUSION: Data are provided on the effect of aluminium hydroxide on key effectors of innate immunity: DC, ML, TLRs and cytokine production. A reasonable administration of this adjuvant was shown to be in association with conjugates of pneumococci synthetic oligosaccharides with a carrier protein.


Assuntos
Adjuvantes Imunológicos/administração & dosagem , Hidróxido de Alumínio/administração & dosagem , Anticorpos Antibacterianos/biossíntese , Antígenos de Bactérias/imunologia , Vacinas Pneumocócicas/imunologia , Pneumonia Pneumocócica/prevenção & controle , Streptococcus pneumoniae/imunologia , Imunidade Adaptativa/efeitos dos fármacos , Hidróxido de Alumínio/imunologia , Animais , Antígenos de Bactérias/administração & dosagem , Antígenos de Bactérias/química , Antígenos de Bactérias/isolamento & purificação , Antígenos CD/genética , Antígenos CD/imunologia , Biomarcadores/metabolismo , Linfócitos T CD8-Positivos/citologia , Linfócitos T CD8-Positivos/efeitos dos fármacos , Linfócitos T CD8-Positivos/imunologia , Citocinas/genética , Citocinas/imunologia , Células Dendríticas/citologia , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Imunidade Inata/efeitos dos fármacos , Imunização , Imunoglobulina G/biossíntese , Injeções Intraperitoneais , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos CBA , Vacinas Pneumocócicas/administração & dosagem , Vacinas Pneumocócicas/química , Pneumonia Pneumocócica/imunologia , Pneumonia Pneumocócica/microbiologia , Streptococcus pneumoniae/química , Equilíbrio Th1-Th2
6.
Artigo em Russo | MEDLINE | ID: mdl-25816514

RESUMO

AIM: Study the protective properties of "Staphylovac-2" vaccinie. MATERIALS AND METHODS: Samples of the vaccine manufactured by SPA "Microgen" based on the developed technology were studied in balb/c mice during 3- and 6-fold immunization schemes. Protective activity of the preparation was determined in experiments with active and passive protection during intraperitoneal infection, seeding of the causative agent from spleen and kidneys during intravenous infection, of animals. RESULTS: In experiments with active protection of mice for both 3- and 6-fold immunization schemes, a significant protective activity of the studied series was determined, compared with the control group of mice. Sera obtained after animal immunization (rabbits, mice) by staphylococcus vaccine had protective properties. A reduction of spleen and kidneys seeding by Staphylococcus aureus in immunized mice compared with the control group was detected in the model of generalized staphylococci infection. CONCLUSION: The preclinical studies carried out with the "Staphylovac-2" vaccine, developed baed on the complex of protective staplylococci antigens, have confirmed the high protective activity of the preparation.


Assuntos
Antígenos de Bactérias/imunologia , Soros Imunes/administração & dosagem , Imunização , Infecções Estafilocócicas/prevenção & controle , Vacinas Antiestafilocócicas/imunologia , Staphylococcus aureus/imunologia , Imunidade Adaptativa/efeitos dos fármacos , Animais , Antígenos de Bactérias/administração & dosagem , Antígenos de Bactérias/química , Carga Bacteriana , Imunidade Inata/efeitos dos fármacos , Injeções Intraperitoneais , Injeções Subcutâneas , Rim/efeitos dos fármacos , Rim/imunologia , Rim/microbiologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Coelhos , Baço/efeitos dos fármacos , Baço/imunologia , Baço/microbiologia , Infecções Estafilocócicas/imunologia , Infecções Estafilocócicas/microbiologia , Vacinas Antiestafilocócicas/administração & dosagem , Vacinas Antiestafilocócicas/química , Staphylococcus aureus/química
7.
Artigo em Russo | MEDLINE | ID: mdl-25816512

RESUMO

AIM: Genotype characteristics of Staphylococcus aureus No 6: strain that is a producer of a protective protein complex. MATERIALS AND METHODS: Features of structure of 9 genes, that code synthesis of pathogenicity factors, of S. aureus--spa, coa, sea, seB, sec, pvl, tst-h, mecA and scc-mecA, that are responsible for synthesis of protein A, coagulase, enterotoxins A, B and C, Panton-Valentine toxin (PVL), heat shock syndrome protein, resistance to methicillin and staphylococci chromosomal cassette, respectively, were studied by amplification in PCR of the respective gene fragments with subsequent conduction of direct sequencing. RESULTS: The S. aureus No 6 strain under study possesses pvl gene fragments, as well as Spa and coagenes, detected in all the studied strains, that belong to t12507 and EMRSA-16 types, respectively. Sea, seb, sec genes responsible for.the synthesis of enterotoxins A, B and C were not detected in it, tst-h, mecA and scc-mecA gene fragments were not present. CONCLUSION: The detection of pvl gene fragment in the strain under study, on the one hand, and protective properties of the secreted protein-containing compound, on the other hand, give evidence in favor of the necessity of further analysis of extracellular proteome of S. aureus No 6.


Assuntos
Regulação Bacteriana da Expressão Gênica , Staphylococcus aureus/genética , Fatores de Virulência/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Toxinas Bacterianas/genética , Toxinas Bacterianas/metabolismo , Coagulase/genética , Coagulase/metabolismo , Enterotoxinas/genética , Enterotoxinas/metabolismo , Exotoxinas/genética , Exotoxinas/metabolismo , Humanos , Leucocidinas/genética , Leucocidinas/metabolismo , Proteínas de Ligação às Penicilinas , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Infecções Estafilocócicas/microbiologia , Proteína Estafilocócica A/genética , Proteína Estafilocócica A/metabolismo , Staphylococcus aureus/isolamento & purificação , Staphylococcus aureus/metabolismo , Fatores de Virulência/metabolismo
8.
Artigo em Russo | MEDLINE | ID: mdl-23805648

RESUMO

AIM: Production of water soluble protein-containing antigens from various strains of S. pneumoniae during cultivation in complete and semi-synthetic culture media as well as selection of strains with cross antigenic activity. MATERIALS AND METHODS: S. pneumoniae 3, 6A, 6B, 14, 10A, 18A, 19A, 19F, 23F serotype strains were cultivated in brain-heart broth and semi-synthetic medium with addition of aminopeptide for 24 hours at 37 degrees C for the production of water soluble antigens. The antigens were obtained by a method of triple water extraction from acetone dried microbial cells. Chemical composition of preparations, electrophoresis mobility of protein-containing components of preparations and cross antigenic activity in gel immune diffusion reaction by using rabbit hyperimmune sera were studied. RESULTS: In studies of 10 pneumococcus strains from various serotypes a method of microbial cell inactivation by acetone was selected that allows to produce preparations with high protein content (25.5 - 53.1%). Electrophoretic separation of the preparations revealed difference in the preparations obtained from various pneumococcus strains in the layout of major protein lines in the 8 - 95 kDa range. The most virulent and immunogenic S. pneumoniae strain that during cultivation in semi-synthetic medium was characterized by intraspecies cross antigenic activity and in gel immune diffusion reacted with all the studied sera against 3, 14, 18C, 23F serotype strains was selected. CONCLUSION: The study resulted in the selection of a technologically simple method of production of pneumococcus antigens with high protein content and showed that only 1 of the studied preparations produced from a virulent strain with poorly expressed S. pneumoniae capsule during cultivation in semi-synthetic medium has the highest cross antigenic activity.


Assuntos
Antígenos de Bactérias , Proteínas de Bactérias , Streptococcus pneumoniae , Animais , Antígenos de Bactérias/química , Antígenos de Bactérias/imunologia , Antígenos de Bactérias/isolamento & purificação , Proteínas de Bactérias/química , Proteínas de Bactérias/imunologia , Proteínas de Bactérias/isolamento & purificação , Meios de Cultura/química , Coelhos , Solubilidade , Streptococcus pneumoniae/química , Streptococcus pneumoniae/crescimento & desenvolvimento , Streptococcus pneumoniae/imunologia , Água/química
9.
Artigo em Russo | MEDLINE | ID: mdl-24605656

RESUMO

AIM: Study intra-species immunogenic activity of antigenic protein-polysaccharide components of S. pneumoniae. MATERIALS AND METHODS: Antigenic components of serotype 3, 6A, 6B, 14, 10A, 18A, 19A, 19F, 23F and unencapsulated S. pneumoniae strains were obtained by water extraction method. Synthetic hexasaccharide--corresponding to the structure of S. pneumoniae serotype 14 capsule polysaccharide repeated unit chain fragment was used as a reference preparation. Molecular mass of antigenic components was determined in SDS-electrophoresis. Antibody titers in blood sera of immunized mice were evaluated by solid-phase EIA method. Protective activity of preparations was studied in mice after 2 immunizations with consequent infection by virulent S. pneumoniae serotype 3 and 6B strains. RESULTS: Preparations from serotype 6A, 6B, 14, 19A, 19F, 23F strains in reaction with anti-microbial sera were characterized by cross serologic activity (IgG titers of 1200 - 12 800). The lowest serologic activity was detected in S. pneumoniae serotype 3 and unencapsulated strain preparations. Conjugate of synthetic hexasaccharide and bovine serum albumin interacted only with homologous antimicrobial sera up to titers of 600 +/- 89.4 and did not react with sera against serotypes 19A and 19E Cross serologic activity of preparations is probably determined by the presence of protein fractions that were detected in SDS-electrophoresis. This is confirmed by high intra-species cross protective activity of preparations from serotype 6B and 10 A strains that protect 90 - 100% of mice from infection by heterologous S. pneumoniae strains. CONCLUSION: Use of strains with cross antigenic and protective activity for production of immunogenic protein-containing fractions with the aim of enchanting and broadening specter of protective activity of vaccine preparations that are constructed based on capsule polysaccharides of S. pneumoniae is appropriate.


Assuntos
Anticorpos Antibacterianos/imunologia , Antígenos de Bactérias/imunologia , Streptococcus pneumoniae/imunologia , Animais , Antígenos de Bactérias/farmacologia , Bovinos , Reações Cruzadas , Camundongos , Camundongos Endogâmicos BALB C , Especificidade da Espécie , Vacinas Estreptocócicas/imunologia , Vacinas Estreptocócicas/farmacologia
10.
Artigo em Russo | MEDLINE | ID: mdl-24605676

RESUMO

AIM: Study antigenic and immunogenic activity of a conjugate of synthetic hexasaccharide related to a S. pneumoniae serotype 14 capsule polysaccharide chain fragment with bovine serum albumin (BSA). MATERIALS AND METHODS: Synthetic glucoconjugate based on BSA protein carrier and hexasaccharide ligand reflecting capsule polysaccharide chain fragment was obtained by using squarate method. Natural polysaccharide-protein complex from S. pneumoniae serotype 14 strain was obtained from cultural fluid supernatant by acetone precipitation. IgG titer against hexasaccharide/capsule polysaccharide was determined in antimicrobial sera and sera of mice immunized with glucoconjugate by EIA method. RESULTS: Immunogenic activity ofglucoconjugate based on BSA protein carriers and synthetic hexasaccharide reflecting S. pneumoniae serotype 14 capsule protein chain fragment was established. After 2 immunizations antibodies against hexasaccharide ligand and BSA were determined in blood sera of mice. Antibody titers against hexasaccharide exceeded the level in intact mice by 4.2 times. BSA in the conjugate did not have effect on production of antibodies against hexasaccharide. CONCLUSION: The developed experimental test-system based on synthetic glucoconjugate is useful for evaluation of level of antibodies against S. pneumoniae serotype 14 in infected and, probably, carriers of bacteria.


Assuntos
Cápsulas Bacterianas , Oligossacarídeos , Polissacarídeos Bacterianos , Streptococcus pneumoniae , Animais , Anticorpos Antibacterianos/imunologia , Cápsulas Bacterianas/química , Cápsulas Bacterianas/imunologia , Bovinos , Camundongos , Camundongos Endogâmicos BALB C , Oligossacarídeos/síntese química , Oligossacarídeos/química , Oligossacarídeos/imunologia , Oligossacarídeos/farmacologia , Polissacarídeos Bacterianos/síntese química , Polissacarídeos Bacterianos/química , Polissacarídeos Bacterianos/imunologia , Polissacarídeos Bacterianos/farmacologia , Soroalbumina Bovina/química , Soroalbumina Bovina/imunologia , Soroalbumina Bovina/farmacologia , Streptococcus pneumoniae/química , Streptococcus pneumoniae/imunologia
11.
Artigo em Russo | MEDLINE | ID: mdl-21913390

RESUMO

AIM: Proof of therapeutic efficacy of a novel type of vaccine with a combination of natural Toll like receptor agonists (TLR) 1/2, 4, 5/6, 9 in infectious and noninfectious human pathology. MATERIALS AND METHODS: Immunovac-VP-4 vaccine, containing antigens of opportunistic microorganisms that are TLR 1/2, 4, 5/6, 9 ligands, was used for experiments and clinical trials. RESULTS: Immunovac-VP-4 activates innate immunity by inducing maturation of dendritic cells with expression of costimulating molecules on their membrane (CD40+, CD80+, CD86+), terminal differentiation molecules--CD83+ and antigen-presenting molecules (MHC class I and II); activates proinflammatory (TNFalpha, IL-6) and regulatory (IL-12, INFgamma) cytokine synthesis that programs T-lymphocyte differentiation by Th1 pathway; increases cytotoxic activity of natural killer cells, inhibits melanoma B16 growth and Lewis lung carcinoma metastasis. Therapeutic effect of Immunovac-VP-4 was evident regardless of pathology by a significant decrease of quantity and severity of recidives, improvement of all clinical parameters, reduction of quantity of administered pharmaceutical preparations including antibiotics and glucocorticosteroids. The rate of intercurrent acute respiratory viral illnesses and their bacterial complications decreased. Immunovac-VP-4 therapy modified course of illness from severe into milder forms. Positive therapeutic effect was 69.2 - 100%. CONCLUSION: High therapeutic effect of vaccine therapy is based on innate immunity activation by combining TLR agonists. Immunovac-VP-4 contains an optimal combination of natural TLR agonists that ensure high therapeutic effect in various nosological forms of infectious and noninfectious human pathology.


Assuntos
Vacinas Bacterianas/imunologia , Vacinas Bacterianas/uso terapêutico , Imunidade Inata , Imunoterapia Ativa/métodos , Receptores Toll-Like/agonistas , Animais , Antígenos de Bactérias/imunologia , Asma/terapia , Carcinoma Pulmonar de Lewis/terapia , Criança , Pré-Escolar , Herpes Genital/terapia , Humanos , Hipersensibilidade ao Látex/terapia , Ativação Linfocitária/imunologia , Melanoma/terapia , Camundongos , Infecções Respiratórias/terapia , Receptores Toll-Like/imunologia , Vacinas Combinadas/imunologia , Vacinas Combinadas/uso terapêutico
12.
Artigo em Russo | MEDLINE | ID: mdl-21446167

RESUMO

AIM: To experimentally assess protective effect of Immunovac-VP-4 vaccine against avian influenza virus H5N2. MATERIALS AND METHODS. Immunization of mice with polycomponent vaccine Immunovac-VP-4 was performed using oral or mucosal route of administration (intranasally, orally, and with combined nasal-oral method). Immunized mice were inoculated intranasally by influenza virus H5N2 adapted for mice. Survival of mice in experimental and control (intact) groups was assessed daily during 14 days. Survival and death rates of mice were determined. Levels of cytokines in sera of mice from both groups were measured by enzyme immunoassay. RESULTS: Half of experimental animals survived after triple subcutaneous administration of vaccine in dose 20 mcg and subsequent intranasal challenge with avian influenza virus H5N2. Single subcutaneous immunization with dose 400 mcg resulted in survival of 80 +/- 12.6% of mice after challenge. Triple intranasal and combined intranasal-oral immunization as well as after triple subcutaneous immunization resulted in survival of half of challenged mice. In control group challenge was lethal for 90 - 100% of mice. Same methods of immunization lead to increase of IL-6, IL-12, IL-15, and IFN-gamma levels. CONCLUSION: Data about significant protective effect after immunization with Immunovac-VP-4 against avian influenza virus H5N2 were obtained. Immunovac-VP-4 administered by mentioned routes activated nasal-associated lymphoid tissue providing first line defense at entry site of influenza infection, which demonstrates need to further study of this vaccine during development of strategy for non-specific prophylaxis of influenza infection.


Assuntos
Citocinas/imunologia , Vírus da Influenza A Subtipo H5N2/imunologia , Vacinas contra Influenza/farmacocinética , Infecções por Orthomyxoviridae/prevenção & controle , Animais , Aves/virologia , Vacinas contra Influenza/imunologia , Influenza Aviária/imunologia , Influenza Aviária/prevenção & controle , Masculino , Camundongos , Camundongos Endogâmicos CBA , Infecções por Orthomyxoviridae/imunologia
13.
Artigo em Russo | MEDLINE | ID: mdl-20737684

RESUMO

AIM: Evaluation of differentiation dynamics and localization of various populations of lymphocytes as well as their expression of TLRs during different methods of administration (intranasal, oral, and subcutaneous) of bacterial ligands of opportunistic microorganisms (Immunovac-VP-4 vaccine) in experiment on mice. MATERIALS AND METHODS: Polycomponent vaccine Immunovac-VP-4 consisting of ligands of 4 opportunistic bacteria was administered to CBA line mice. Groups of mice were immunized orally, subcutaneously, or intranasally. Number of mononuclear leukocytes, as well as levels of cytokines, lymphocytic antigens, and cytotoxic activity of cells were measured. RESULTS: It was demonstrated that modification of immunophenotype of lymphocytes and cytotoxic activity of NK cells depends from route of administration of Immunovac-VP-4 because the most intensive activation of cells occurred in organs proximal to place of vaccine application. However, already 1 day after administration of vaccine there was intensive exchange between lymphoid cells not only in nasal associated lymphoid tissue, bronchial associated lymphoid tissue, and gut associated lymphoid tissue but also in spleen that points to integration of fine mechanisms of mucosal and systemic immune response regulation. CONCLUSION: Development of noninvasive methods of vaccination is an optimal way of safe and mass prevention of infectious diseases.


Assuntos
Vacinas Bacterianas/administração & dosagem , Vacinas Bacterianas/imunologia , Tecido Linfoide/imunologia , Administração Intranasal , Administração Oral , Animais , Diferenciação Celular , Linhagem Celular , Citocinas/análise , Infecções por Escherichia coli/imunologia , Humanos , Imunização , Injeções Subcutâneas , Células Matadoras Naturais/imunologia , Infecções por Klebsiella/imunologia , Leucócitos Mononucleares/imunologia , Ligantes , Camundongos , Camundongos Endogâmicos CBA , Infecções por Proteus/imunologia , Receptores de Antígenos de Linfócitos T/biossíntese , Infecções Estafilocócicas/imunologia , Vacinas Combinadas/administração & dosagem , Vacinas Combinadas/imunologia
14.
Artigo em Russo | MEDLINE | ID: mdl-20795383

RESUMO

AIM: Assessment of therapeutic effect and immunologic parameters during use of Immunovac vaccine for complex treatment of chronic forms of pyoderma. MATERIALS AND METHODS: Ninety-five patients with different clinical forms of chronic pyoderma (furunculosis, hydradenitis, chronic ulcerative and ulcerative-vegetans pyoderma, folliculitis, impetigo etc.) were studied. Fifty-nine patients received immunotherapy with Immunovac vaccine together with basic therapy and 36 patients comprised control group treated only with basic therapy. Studied immunologic parameters were as follows: assessment of functional activity of lymphocytes, determination of lymphocyte subpopulations by flow cytometry, total immunoglobulins classes A, G, M by radial immunoduffusion, affinity of antibodies by enzyme immunoassay, levels of IFNalpha and IFNgamma. RESULTS: Use of Immunovac vaccine in complex treatment of patients with chronic forms of pyoderma enhanced clinical effect of basic therapy, which expressed in decrease of severity and frequency of disease relapses irrespective to clinical form and severity of pyoderma. Therapeutic effect during use of Immunovac vaccine amounted 84.7%, whereas in control group it was 41.6% after 12 months of follow-up. Increase of functional activity of neutrophils, subpopulation of lymphocytes with markers CD4+, CD8+, CD72+, affinity of antibodies as well as induced production of IFNalpha and IFNgamma was revealed. Correction of immunologic parameters correlated with positive results of patients treatment. CONCLUSION: Inclusion of bacterial polycomponent vaccine Immunovac in complex treatment of patients with chronic pyoderma promotes enhancement of therapeutic effect of basic therapy and correction of immunologic parameters.


Assuntos
Vacinas Bacterianas/uso terapêutico , Imunoterapia , Pioderma/terapia , Adolescente , Adulto , Idoso , Antígenos CD/biossíntese , Antígenos de Diferenciação de Linfócitos B/biossíntese , Antígenos CD4/biossíntese , Antígenos CD8/biossíntese , Doença Crônica , Humanos , Imunoglobulinas/sangue , Imunoglobulinas/imunologia , Interferon-alfa/sangue , Interferon gama/sangue , Contagem de Linfócitos , Pessoa de Meia-Idade , Pioderma/sangue , Pioderma/imunologia , Prevenção Secundária , Linfócitos T/imunologia , Linfócitos T/metabolismo , Resultado do Tratamento , Vacinas Combinadas/uso terapêutico
15.
Artigo em Russo | MEDLINE | ID: mdl-20465001

RESUMO

AIM: Assessment of characteristics of antigenic complexes of Staphylococcus aureus vaccine strains in different cultivation conditions. MATERIALS AND METHODS: S. aureus vaccine strains (No. 5, 9, 1986, 1991) were grown in liquid nutrient media--full value and semi-synthetic--as well as on solid medium. Reactor cultivation was performed in the fermenter ANKUM-2M. Complex of antigens were obtained by water extraction method applied to staphylococcal biomass inactivated with acetone and assessed by common methods on protein and carbohydrate content; specific activity was assessed by minimal inhibitory dose in passive hemagglutination inhibition assay. Study of acute toxicity was performed on outbred mice. RESULTS: Using strain no. 1991, model of reactor cultivation in full value medium with separation of biomass by microfiltration was validated on the basis of biomass and semiproduct of antigenic complex (acetone powder) yield as well as productivity of biomass cumulation. Study of antigenic complexes obtained from biomass of 4 strains during reactor cultivation compared with complexes extracted from cultures grown on solid medium revealed increased protein and decreased carbohydrate content but similar specific activity. It was demonstrated that complex of antigens obtained from cultures grown either by reactor cultivation or on solid medium were non-toxic. CONCLUSION: New technology for manufacturing staphylococcal complex of antigens with reactor cultivation of vaccine strains in full value medium with subsequent purification of antigenic complex from the biomass by microfiltration was developed. Results of the study demonstrated the usefulness of the developed technology for both further studies on a cellular staphylococcal vaccine and manufacture of staphylococcal component of "Immunovac" vaccine.


Assuntos
Antígenos de Bactérias/isolamento & purificação , Reatores Biológicos , Microbiologia Industrial/métodos , Infecções Estafilocócicas/prevenção & controle , Vacinas Antiestafilocócicas/isolamento & purificação , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus aureus/imunologia , Acetona , Animais , Antígenos de Bactérias/imunologia , Meios de Cultura , Microbiologia Industrial/normas , Camundongos , Vacinas Antiestafilocócicas/administração & dosagem , Vacinas Antiestafilocócicas/imunologia
16.
Artigo em Russo | MEDLINE | ID: mdl-20218346

RESUMO

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.


Assuntos
Vacinas Bacterianas/administração & dosagem , Linfonodos/imunologia , Baço/imunologia , Receptores Toll-Like/biossíntese , Administração Intranasal , Administração Oral , Animais , Vacinas Bacterianas/imunologia , Linhagem Celular , Imunização , Intestino Delgado/imunologia , Masculino , Camundongos , Camundongos Endogâmicos CBA , Mucosa/imunologia , Sistema Respiratório/imunologia , Vacinas Acelulares/administração & dosagem , Vacinas Acelulares/imunologia
17.
Artigo em Russo | MEDLINE | ID: mdl-21381375

RESUMO

AIM: Assessment of prophylactic effect of Immunovac-VP-4 against acute respiratory diseases (ARDs) in collectives of children. MATERIALS AND METHODS: Immunovac-VP-4 was used for prophylaxis of ARDs in communities of children (daycare centers, boarding school). During 3 consecutive controlled studies performed during peak incidence of epidemic influenza and ARD (1st study) and during pre-epidemic period (2nd and 3rd studies) the incidence of ARDs was studied in groups of vaccinated and non-vaccinated children. Criteria for assessment of ARDs incidence and their severity were as follows: number of ARD episodes per child, number of children with recurrent episodes of ARD during period of follow-up, duration of ARDs, and number of children with bacterial complications (bronchitis, otitis, etc.). Effect of intervention on immunologic parameters was assessed on levels of sIgA, IgG, IgA, and antibody titers in saliva. Schedule of vaccine administration was 3 daily intranasal and 6 - 9 oral administration of the vaccine with 3 - 4 days interval. RESULTS: Results of all three studies were virtually the same: decrease of number of ARD episodes and their duration; 3.1-fold reduction of number of children with recurrent infections during 14 months of follow-up, and 10.9--12-fold reduction during first 7 months of follow-up; decrease of bacterial complications rate (1.9% in children in intervention group, 11.9%--in control group). Increase of total immunoglobulin level and antibody titers to antigenic components of the vaccine from low baseline level was observed in saliva of vaccinated children. CONCLUSION: Immunovac-VP-4 results in enhanced and prolonged prophylactic effect on ARDs incidence and recommended for building of optimal system of prevention of polyetiologic complex of ARDs.


Assuntos
Antígenos de Bactérias/administração & dosagem , Fatores Imunológicos/administração & dosagem , Infecções Respiratórias/prevenção & controle , Vacinação , Doença Aguda , Administração Intranasal , Administração Oral , Anticorpos Antibacterianos/análise , Antígenos de Bactérias/imunologia , Criança , Creches , Pré-Escolar , Escherichia coli/imunologia , Humanos , Imunoglobulina A/análise , Imunoglobulina A Secretora/análise , Imunoglobulina G/análise , Fatores Imunológicos/imunologia , Klebsiella pneumoniae/imunologia , Proteus vulgaris/imunologia , Infecções Respiratórias/imunologia , Saliva/imunologia , Staphylococcus aureus/imunologia
18.
Artigo em Russo | MEDLINE | ID: mdl-19338235

RESUMO

Experimental study of adjuvant effect of recombinant mycobacterial heat-shock protein rHSP70 on immune response to antigens of opportunistic microorganisms was performed. Therapeutic poly-component vaccine Immunovac-VP-4, containing ligands for binding with Toll-like receptors (TLRs) 2, 4 and 9, was used as a source of bacterial antigens. Obtained data showed that administration of rHSP70 mixed with bacterial antigens of opportunistic microorganisms leads to maturation of mice dendritic cells obtained from bone marrow precursors, increased expression of TLRs 2, 4, 9 on their surface and production of cytokines IL-1beta, IL-6, IL-10, TNFalpha, as well as to activation of innate immunity in experiments in vivo resulting to increased resistance of mice to experimental Salmonella infection. Simultaneous administration of rHSP70 with bacterial antigens increased titers of antibodies to vaccine's antigenic components in direct hemagglutination reaction. Thus, adjuvant effect of rHSP70 was confirmed on the used model as well as increase of stimulation of innate and adaptive immunity was shown.


Assuntos
Adjuvantes Imunológicos/administração & dosagem , Proteínas de Bactérias/imunologia , Vacinas Bacterianas/imunologia , Proteínas de Choque Térmico HSP70/imunologia , Imunidade Inata , Vacinas Combinadas/imunologia , Animais , Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/imunologia , Proteínas de Bactérias/administração & dosagem , Vacinas Bacterianas/administração & dosagem , Citocinas/biossíntese , Células Dendríticas/imunologia , Proteínas de Choque Térmico HSP70/administração & dosagem , Imunidade Ativa , Injeções Intraperitoneais , Camundongos , Camundongos Endogâmicos CBA , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/imunologia , Infecções por Salmonella/prevenção & controle , Receptores Toll-Like/metabolismo , Vacinas Combinadas/administração & dosagem
19.
Artigo em Russo | MEDLINE | ID: mdl-18822497

RESUMO

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.


Assuntos
Antígenos de Bactérias/imunologia , Proteínas de Bactérias/imunologia , Infecções por Klebsiella/prevenção & controle , Klebsiella pneumoniae/imunologia , Infecções Pneumocócicas/prevenção & controle , Streptococcus pneumoniae/imunologia , Animais , Antígenos de Bactérias/administração & dosagem , Antígenos de Bactérias/metabolismo , Proteínas de Bactérias/administração & dosagem , Proteínas de Bactérias/metabolismo , Meios de Cultura , Soros Imunes/administração & dosagem , Imunização Passiva , Klebsiella pneumoniae/metabolismo , Camundongos , Camundongos Endogâmicos CBA , Peso Molecular , Streptococcus pneumoniae/metabolismo , Vacinação
20.
Artigo em Russo | MEDLINE | ID: mdl-19186542

RESUMO

In the overwhelming majority of countries inactivated vaccines, which form mainly humoral immunity, are used for prevention of influenza. The objective of the study was to assess the combined effect of inactivated influenza vaccine and chitozan on cellular immunity in CBA line mice. Intramuscular administration of 2 doses (with 4 week interval) of inactivated influenza vaccine and chitozan resulted in increased cytotoxic activity of splenic NK cells against NK-sensitive cell line K562 as well as in increased proliferative activity of mononuclear leukocytes, and numbers of CD3 T-lymphocytes, NKT cells, B-lymphocytes in animals' spleens. Combination of inactivated influenza vaccine with chitozan modulated the number MHC II-expressing cells by eliminating the increased reactivity of immune system cells as well as increased the number of MHC I-expressing cells. This point on the activation of cellular properties, which recognize intracellular pathogens, and thus on activation of both humoral and cellular factors of immune response. It can be proposed that inclusion of chitozan in the vaccine allows to modulate switching of the immune response from Th-2 to Th-1 type.


Assuntos
Adjuvantes Imunológicos/administração & dosagem , Quitosana/administração & dosagem , Quitosana/imunologia , Vacinas contra Influenza/imunologia , Influenza Humana/prevenção & controle , Leucócitos Mononucleares/imunologia , Vacinas de Produtos Inativados/imunologia , Animais , Complexo CD3/metabolismo , Humanos , Imunidade Celular , Esquemas de Imunização , Vacinas contra Influenza/administração & dosagem , Influenza Humana/sangue , Influenza Humana/imunologia , Injeções Intramusculares , Células Matadoras Naturais/imunologia , Ativação Linfocitária , Complexo Principal de Histocompatibilidade/imunologia , Camundongos , Camundongos Endogâmicos CBA , Baço/imunologia , Linfócitos T/imunologia , Linfócitos T/metabolismo , Linfócitos T Citotóxicos , Vacinas de Produtos Inativados/administração & dosagem
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