RESUMEN
The transcription factor NF-κB is a key factor in the activation of immune responses; it is in turn activated by pattern recognition receptors, such as TLR and NLR receptors. The search for ligands activating innate immunity receptors is an important scientific problem due to the possibility of their use as adjuvants and immunomodulators. In this study the effect of recombinant Pseudomonas aeruginosa OprF proteins and a toxoid (a deletion atoxic form of exotoxin A) on the activation of TLR4, TLR9, NOD1, and NOD2 receptors has been investigated. The study was carried out using free and co-adsorbed on Al(OH)3 P. aeruginosa proteins and eukaryotic cells encoding these receptors and having NF-κB-dependent reporter genes. The enzymes encoded by the reported genes are able to cleave the substrate with the formation of a colored product, the concentration of which indicates the degree of receptor activation. It was found that free and adsorbed forms of the toxoid were able to activate the TLR4 surface receptor for lipopolysaccharide. OprF and the toxoid activated the intracellular NOD1 receptor, but only in the free form. This may be due to the fact that the cell lines used were not able to phagocytize aluminum hydroxide particles with protein adsorbed on them.
Asunto(s)
FN-kappa B , Pseudomonas aeruginosa , Células Eucariotas , Receptor Toll-Like 4/genética , Toxoides , Adyuvantes Inmunológicos , Proteínas Recombinantes/genéticaRESUMEN
Pneumolysin (Ply) is a target for the development of serotype-independent pneumococcal vaccines, an important condition for the efficacy of which is their ability to activate innate immunity with the subsequent formation of adaptive immunity. In this study, the ability of recombinant full-length Ply (rPly) of pneumococci to induce TLR expression and maturation of dendritic cells generated from mouse bone marrow was evaluated. It was shown that rPly in vitro increased the number of dendritic cells expressing Toll-like receptor 4 (TLR4) on the membrane. rPly caused maturation of dendritic cells generated from mouse bone marrow, which manifested in a decrease in the number of progenitor cells (CD34), an increase in the number of cells expressing the adhesion molecule CD38, costimulatory molecules CD80 and CD86, molecules of terminal differentiation of dendritic cells CD83, as well as molecules of antigenic presentation of the major histocompatibility complex class II.
Asunto(s)
Streptococcus pneumoniae , Estreptolisinas , Receptor Toll-Like 4 , Ratones , Animales , Streptococcus pneumoniae/genética , Receptor Toll-Like 4/genética , Receptor Toll-Like 4/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/farmacología , Proteínas Bacterianas/metabolismo , Células DendríticasRESUMEN
The effects of various sulfated polysaccharides of brown algae Fucus evanescens, Saccharina cichorioides and Saccharina japonica on the morphofunctional changes of dendritic cells have been investigated using flow cytometry and phase-contrast microscopy. The dendritic cells are characterized by larger sizes, vacuolated cytoplasm, eccentrically located nucleus, and also by the presence of numerous cytoplasmic pseudopodia of various shapes. They express surface markers, indicating their maturation (CD83, CD11c, HLA-DR, CD86). Increased production of immunoregulatory (IL-12) and proinflammatory TNF-a, IL-6) cytokines (by dendritic cells polarizes the development of the Th-1 type immune response.
Asunto(s)
Células Dendríticas/efectos de los fármacos , Expresión Génica/efectos de los fármacos , Monocitos/efectos de los fármacos , Phaeophyceae/química , Polisacáridos/farmacología , Seudópodos/efectos de los fármacos , Antígenos CD/genética , Antígenos CD/inmunología , Antígeno B7-2/genética , Antígeno B7-2/inmunología , Antígeno CD11c/genética , Antígeno CD11c/inmunología , Secuencia de Carbohidratos , Diferenciación Celular , Tamaño de la Célula , Células Dendríticas/inmunología , Células Dendríticas/ultraestructura , Citometría de Flujo , Antígenos HLA-DR/genética , Antígenos HLA-DR/inmunología , Humanos , Inmunidad Innata , Inmunoglobulinas/genética , Inmunoglobulinas/inmunología , Interleucina-12/genética , Interleucina-12/inmunología , Interleucina-6/genética , Interleucina-6/inmunología , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/inmunología , Monocitos/inmunología , Monocitos/ultraestructura , Polisacáridos/aislamiento & purificación , Cultivo Primario de Células , Seudópodos/ultraestructura , Ésteres del Ácido Sulfúrico/química , Balance Th1 - Th2/efectos de los fármacos , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/inmunología , Antígeno CD83RESUMEN
AIM: Study immunologic phenotype of lymphocytes in the process of therapy of topic form of recombinant interferon-α2b during respiratory infections in pregnant. MATERIALS AND METHODS: 74 pregnant women from 14 weeks of gestation took part in the study, among them 55 - within 24 hours with symptoms of acute respiratory infection (ARI) of light and me- dium, severe course of infection, who do not need hospitalization. Group I - 34 pregnant womenwithARI receivingbasic therapywith human recombinant interferon-(α2b in gelform. Group II - 21 pregnant with ARI receiving only basic therapy. Control group had 19 pregnant women without signs of ARI. Relative content of principle lymphocyte subpopulations was studied by flow cytofluorimetry: CD3+, CD3+CD4+, CD3+CD8+, CD3-CD19+, CD3- CD16+56+, CD3-CD8+; immune regulatory indexwas calculated inblood within 24 hours from the onset of the disease and 8 - 10 days later. RESULTS: A disbalance of lymphocyte sub- populations was noted in pregnant women with light or medium severity course of acute respiratory infections, that was characterized by an increased content of CD3-CD16+56+ and CD3+CD8+, as well as a reduced content of CD3+ and CD3+CD8+. Inclusion of a topical form of recombinant interferon-α2b during the first days of development ofthe disease has a systemic effect on cell immunity and results in normalization of subpopulation compo- sition of blood lymphocytes that is characteristic for physiological course of pregnancy. CONCLUSION: Administration of topic form of recombinant interferon in pregnant with light or medium severity of ARI can be accompanied by activation of factors of innate and adaptive immunity.
Asunto(s)
Inmunidad Adaptativa/efectos de los fármacos , Inmunidad Innata/efectos de los fármacos , Interferón alfa-2/administración & dosificación , Complicaciones Infecciosas del Embarazo , Infecciones del Sistema Respiratorio , Administración Tópica , Adulto , Antígenos CD/inmunología , Femenino , Humanos , Linfocitos/inmunología , Embarazo , Complicaciones Infecciosas del Embarazo/tratamiento farmacológico , Complicaciones Infecciosas del Embarazo/inmunología , Infecciones del Sistema Respiratorio/tratamiento farmacológico , Infecciones del Sistema Respiratorio/inmunologíaRESUMEN
AIM: Study the effect of Vaxigrip split, Influvac subunit and Grippol plus immune-adjuvanted vaccines on the content of myeloid (mDC) and plasmacytoid (pDC) dendritic cells (DC) in blood of vaccinated healthy women. Materials andmethods. Blood of 30 healthy women aged 18-50 years was studied at days 7 and 30 after the vaccination. pDC (CD14+CD16-/CD85k(ILT3)-PE/ CD123-PC5) and mDC (CD14+CD-16-/CD85k(ILT3)-PE/CD33 -PC5) immune phenotyping was carried out using mAbs (Beckman Coulter, France) and flow cytometer Cytomix FC-500 (Beckman Coulter, USA). RESULTS: Use of unadjuvanted vaccines Vaxigrip and Influvac resulted in an increase of the numbers of mDC and pDC (p<0.05) in blood of the vaccinated only at day 7 of the observation. Grippol resulted in a more significant (2.2 - 3.6 times, p<0.05) increase of DC subpopulations (compared with unadjuvanted vaccines) at both day 7 and a month after the vaccination. CONCLUSION: Influenza vaccination activated innate effectors - the first component on the way of infection penetration - dendritic cells of both myeloid and lymphoid origin. Wherein, a more pronounced and prolonged effect of such activation is observed when immune-adjuvanted vaccine is used compared with subunit and split vaccines.
Asunto(s)
Células Sanguíneas/inmunología , Células Dendríticas/inmunología , Inmunidad Innata/efectos de los fármacos , Vacunas contra la Influenza/administración & dosificación , Adolescente , Adulto , Antígenos CD/inmunología , Células Sanguíneas/citología , Células Dendríticas/citología , Femenino , Humanos , Vacunas contra la Influenza/inmunologíaRESUMEN
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.
Asunto(s)
Anticuerpos Antibacterianos/inmunología , Cápsulas Bacterianas/inmunología , Inmunoglobulina G/inmunología , Vacunas Neumococicas/inmunología , Polisacáridos Bacterianos/inmunología , Streptococcus pneumoniae/inmunología , Vacunación , Animales , Ratones , Ratones Endogámicos BALB C , Vacunas Neumococicas/farmacología , Polisacáridos Bacterianos/farmacologíaRESUMEN
AIM: Study subpopulation structure of lymphocytes in healthy individuals under the effect of various influenza vaccines in an in vitro system. MATERIALS AND METHODS: Evaluation of immune- phenotype features of PBMCs, activated in vitro by immune-adjuvanted and unadjuvanted vaccines against influenza in healthy individuals, was carried out by using flow cytometry method. RESULTS: Grippol plus vaccine caused a more pronounced stimulating effect compared with subunit and split-vaccines on NK-cells, cells with markers of early activation CD45/CD25, induced the quantity of natural regulatory cells (CD4/CD25/Foxp3), increase of the number of B-cells and reduced the amount of cell types with apoptosis marker CD45/CD95. CONCLUSION: Immune-adjuvanted vaccine Grippol plus induced formation of effectors of both innate and adaptive immunity and possessed the most powerful potential regarding activation of various types of immune-competent cells compared with unadjuvanted vaccines.
Asunto(s)
Inmunidad Adaptativa , Antígenos CD/inmunología , Inmunidad Innata , Vacunas contra la Influenza/administración & dosificación , Linfocitos T Reguladores/inmunología , Adolescente , Adulto , Femenino , Citometría de Flujo , Humanos , Vacunas contra la Influenza/inmunología , Persona de Mediana EdadRESUMEN
AIM: Study innate and adaptive immunity in patients with migrating erythema, clinical effectiveness ofcombined therapy using Immunovac vaccine and dynamics of immunologic parame- ters as a result of the therapy. MATERIALS AND METHODS: 37 adult patients with migrating erythema were examined. The patients were divided into 2 groups: 1st gr. (14 individuals) - Immunovac by intranasal-subcutaneous method against the background of basic therapy; 2nd gr. (23 individuals) - 200 mg/day doxycycline therapy for 21 days. Phagocytic activity of blood neutrophils; TLRs expression on mononuclear leukocytes of peripheral blood (PBML) and skin cells in foci by flow cytometry with mAT against TLR2, 3, 4, 5, 6, 7, 8, 9 using flow cytometer FC-500; subpopulation composition of peripheral blood lymphocytes; levels of pro-, anti-inflammation and regulatory cytokines in blood sera by EIA method; IgG, 1gM, IgA in blood sera were studied in patients before treatment and 1 month after therapy. RESULTS: A high level of TLR2, 4, 7, 8 on skin cells in foci, TLR2, 4 - on blood cells; low content of CD95+ and CD25+, high-level of serum IL-lb, IL-2 and IL-4, an increase of general IgE level was detected in patients'with migrating erythema. Immunovac facilitated an increase of CD95+ and CD25+, IFN-y synthesis, reduced the level of general IgE in a more pronounced way than basic therapy. CONCLUSION: Inclusion of Immunovac into therapy of patients With migrating erythema facilitates increase of clinical effectiveness and correlates with correction of immunologic disorders.
Asunto(s)
Doxiciclina/administración & dosificación , Eritema , Vacunas contra Enfermedad de Lyme/administración & dosificación , Enfermedad de Lyme , Neutrófilos/inmunología , Adulto , Anticuerpos Antibacterianos/inmunología , Citocinas/inmunología , Eritema/tratamiento farmacológico , Eritema/inmunología , Femenino , Humanos , Enfermedad de Lyme/tratamiento farmacológico , Enfermedad de Lyme/inmunología , Masculino , Persona de Mediana Edad , Fagocitosis/efectos de los fármacos , Receptores Toll-Like/inmunologíaRESUMEN
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.
Asunto(s)
Disacáridos/inmunología , Infecciones Neumocócicas/inmunología , Streptococcus pneumoniae/inmunología , Vacunas Conjugadas/inmunología , Animales , Anticuerpos Antibacterianos/inmunología , Cápsulas Bacterianas/inmunología , Bovinos , Disacáridos/síntesis química , Epítopos/inmunología , Humanos , Ratones , Infecciones Neumocócicas/prevención & control , Sensibilidad y Especificidad , Serogrupo , Albúmina Sérica Bovina/administración & dosificación , Albúmina Sérica Bovina/inmunología , Streptococcus pneumoniae/patogenicidad , Vacunación , Vacunas Conjugadas/administración & dosificaciónRESUMEN
The effect of sulfated polysaccharide fucoidan from the brown alga Laminaria japonica on morphological characteristics of mouse lymphoid organs, subpopulations of spleen mononuclear leukocytes, cytokine production and cytotoxic activity of splenocytes has been investigated. Fucoidan promoted activation and proliferation of lymphoid hematopoietic cells in primary and secondary immunogenesis bodies, increased expression of markers CD19, NK, NKT, CD25, MHC II, TCR, TLR2 and TLR4, the cytotoxic activity of splenocytes and production of immunoregulatory and proinflammatory cytokines (IL- 2, IL-12, IFN-g, TNF-a, IL-6). This suggests activation of effector mechanisms of innate immunity and adaptive immune responses via the Th-1 type.
Asunto(s)
Laminaria/química , Leucocitos Mononucleares/efectos de los fármacos , Ganglios Linfáticos/efectos de los fármacos , Extractos Vegetales/farmacología , Polisacáridos/farmacología , Bazo/efectos de los fármacos , Timo/efectos de los fármacos , Animales , Antígenos CD19/genética , Antígenos CD19/metabolismo , Médula Ósea/anatomía & histología , Médula Ósea/efectos de los fármacos , Citocinas/genética , Citocinas/metabolismo , Leucocitos Mononucleares/metabolismo , Ganglios Linfáticos/anatomía & histología , Ratones , Extractos Vegetales/química , Polisacáridos/química , Bazo/anatomía & histología , Timo/anatomía & histología , Receptores Toll-Like/genética , Receptores Toll-Like/metabolismoRESUMEN
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.
Asunto(s)
Glicoconjugados/farmacología , Polisacáridos/química , Streptococcus pneumoniae/química , Animales , Secuencia de Carbohidratos , Ratones , Ratones Endogámicos BALB C , Datos de Secuencia MolecularRESUMEN
The minimal vaccination dose of tumor cells was determined on experimental models. The effectiveness of antitumor immunotherapy with activated natural killer cells or dendritic cell vaccine (monotherapy or combined treatment) was evaluated in vivo. The inhibition of tumor growth was more pronounced after combination therapy with activated natural killer cells and dendritic cell-based vaccine. Our results indicate that the effectiveness of antitumor immunotherapy increases in simultaneous modulation of both immune components (innate and adaptive immunity).
Asunto(s)
Vacunas contra el Cáncer/uso terapéutico , Melanoma Experimental/terapia , Animales , Vacunas contra el Cáncer/inmunología , Células Dendríticas/inmunología , Células Asesinas Naturales/inmunología , Masculino , Melanoma Experimental/inmunología , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos CBARESUMEN
The article reports survey data on 23 patients with erythemas, including 19 patients with herpes-associated erythema multiforme (HAEM) and 4 patients with Darier's erythema annulare centrifugum (DEAC). Patients in the initial state (baseline) and after two weeks of therapy with proteflazid were characterized by measuring the levels of Toll-like receptor (TLR) expression in peripheral blood mononuclear cells (PBMC) and in epithelial cells of the throat and the skin. The TLR expression in PBMC and skin was assessed by flow cytometry with monoclonal antibodies (ICA) (Caltag Laboratories, USA; Hycult Biotech, Netherlands) against relevant antigens. In addition, patients were also characterized by the content of subpopulations of lymphocytes expressing surface markers CD3, CD4, CD8, CD16, CD21, CD23, CD72, CD25, and HLA-DR in the peripheral blood, which was measured by flow cytometry. The therapy with proteflazid in patients with both HAEM and DEAC led to normalization of the level of both T-cell and B-cell immunity, which was manifested by an increase in the total number of lymphocytes, CD3+, CD4+, CD21+, and CD72+. Measurements of the dynamics of TLR expression in the course of immunotherapy showed an increase in the number of TLR 2, 3, 4, 7, 8, and 9 in PBMC (which was especially pronounced for TLR2) and in epithelium of the pharyngeal mucosa and skin (increased expression of TLR3, 7, and 9).
Asunto(s)
Antivirales/uso terapéutico , Células Epiteliales/efectos de los fármacos , Eritema Multiforme/tratamiento farmacológico , Eritema/tratamiento farmacológico , Expresión Génica/efectos de los fármacos , Herpes Simple/tratamiento farmacológico , Mucosa Bucal/efectos de los fármacos , Enfermedades Cutáneas Genéticas/tratamiento farmacológico , Receptores Toll-Like/genética , Adolescente , Adulto , Antígenos CD/genética , Antígenos CD/inmunología , Linfocitos B/efectos de los fármacos , Linfocitos B/inmunología , Linfocitos B/metabolismo , Células Epiteliales/inmunología , Células Epiteliales/metabolismo , Eritema/complicaciones , Eritema/genética , Eritema/inmunología , Eritema Multiforme/complicaciones , Eritema Multiforme/genética , Eritema Multiforme/inmunología , Femenino , Flavonoides/uso terapéutico , Citometría de Flujo , Glicósidos/uso terapéutico , Antígenos HLA-DR/genética , Antígenos HLA-DR/inmunología , Herpes Simple/complicaciones , Herpes Simple/genética , Herpes Simple/inmunología , Humanos , Inmunidad Celular , Inmunidad Innata , Masculino , Mucosa Bucal/inmunología , Mucosa Bucal/metabolismo , Faringe/efectos de los fármacos , Faringe/inmunología , Faringe/metabolismo , Piel/efectos de los fármacos , Piel/inmunología , Piel/metabolismo , Enfermedades Cutáneas Genéticas/complicaciones , Enfermedades Cutáneas Genéticas/genética , Enfermedades Cutáneas Genéticas/inmunología , Linfocitos T/efectos de los fármacos , Linfocitos T/inmunología , Linfocitos T/metabolismo , Receptores Toll-Like/agonistas , Receptores Toll-Like/inmunologíaRESUMEN
AIM: Study the production of cytokines in mice during vaccination with polycomponent Immunovac-VP-4 vaccine containing TLR ligands with various administration methods. MATERIALS AND METHODS: Immunovac-VP-4 was administered to mice by subcutaneous, intranasal or per oral methods. The preparation was administered nasally at a single dose of 500 microg in the volume of 30 microl. Per oral single dose was 2000 microg in the volume of 0.5 ml. 200 microg of the preparation was administered subcutaneously. Cytokines in blood sera were determined by EIA 8 hours after the administration of the vaccine. RESULTS: In mice 8 hours afterthe single administration of Immunovac-VP-4 the levels of IL-1beta, IL-6, IL- 12, IL-5 increased significantly. However their concentration differed depending on the method of administration. The most active expression of cytokines was observed during subcutaneous administration. The indexes of cytokine expression were significantly higher (p < 0.05) than during non-parenteral administration methods. CONCLUSION: Mucosal application methods along with parenteral were established to be able to activate effector mechanisms of immune repose with its consequent polarization by Th1/Th2 pathways. These mechanisms lay the groundwork for development of antigen-specific immune responses against antigens/pathogens.
Asunto(s)
Vacunas Bacterianas/farmacología , Citocinas/inmunología , Regulación de la Expresión Génica/efectos de los fármacos , Animales , Bacterias/inmunología , Vacunas Bacterianas/inmunología , Citocinas/biosíntesis , Relación Dosis-Respuesta Inmunológica , Regulación de la Expresión Génica/inmunología , Ratones , Ratones Endogámicos CBA , Células TH1/inmunología , Células TH1/metabolismo , Células Th2/inmunología , Células Th2/metabolismoRESUMEN
AIM: Study the features of cytokine profile in patients with exudative erythema multiforme (EME) and dynamics of basal level of pro-inflammatory and anti-inflammatory cytokines during immunotherapy. MATERIALS AND METHODS: 39 adult patients with erythema multiforme were examined. The patients were split into groups based on therapy variant. One group (14 individuals) received Immunovac-VP-4 against the background of basic therapy; the other (12 individuals)--cagocel against the background of basic therapy; comparison group (13 individuals) received only basic therapy; 15 individuals composed a group of healthy individuals. All the patients had the level of pro-, anti-inflammatory and regulatory cytokines determined in blood sera by solid-phase EIA method by using Biosource (Austria) test-systems at the beginning of the study and after the therapy. RESULTS: In patients with exudative erythema multiforme Immunovac-VP-4 therapy facilitated a significant (p < 0.05) increase of serum IFN-gamma level, insignificant (p > 0.05) increase of IL-1beta and decrease of IL-17. Whereas cagocel lead to an increase of IL-4 (p > 0.05), IL-2, IFN-gamma (p < 0.05) and decrease of TGF-beta and IL-12 (p < 0.05). At the same time basic therapy facilitated a significant increase of IL-5 and decrease of IL-6, IL-12, IFN-gamma. CONCLUSION: Immunovac-VP-4 facilitates the increase of secretion of IFN-gamma, IL-1beta against the background of TGF-beta that facilitates normalization of cooperation of cells in immune response including against viral infections, and thereby influencing the trigger factor in EME patients.
Asunto(s)
Citocinas/sangre , Eritema Multiforme/sangre , Eritema Multiforme/terapia , Inmunoterapia , Adolescente , Adulto , Antiinflamatorios/administración & dosificación , Citocinas/inmunología , Eritema Multiforme/inmunología , Femenino , Gosipol/administración & dosificación , Gosipol/análogos & derivados , Humanos , Masculino , Persona de Mediana EdadRESUMEN
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.
Asunto(s)
Adyuvantes Inmunológicos/administración & dosificación , Hidróxido de Aluminio/administración & dosificación , Anticuerpos Antibacterianos/biosíntesis , Antígenos Bacterianos/inmunología , Vacunas Neumococicas/inmunología , Neumonía Neumocócica/prevención & control , Streptococcus pneumoniae/inmunología , Inmunidad Adaptativa/efectos de los fármacos , Hidróxido de Aluminio/inmunología , Animales , Antígenos Bacterianos/administración & dosificación , Antígenos Bacterianos/química , Antígenos Bacterianos/aislamiento & purificación , Antígenos CD/genética , Antígenos CD/inmunología , Biomarcadores/metabolismo , Linfocitos T CD8-positivos/citología , Linfocitos T CD8-positivos/efectos de los fármacos , Linfocitos T CD8-positivos/inmunología , Citocinas/genética , Citocinas/inmunología , Células Dendríticas/citología , Células Dendríticas/efectos de los fármacos , Células Dendríticas/inmunología , Inmunidad Innata/efectos de los fármacos , Inmunización , Inmunoglobulina G/biosíntesis , Inyecciones Intraperitoneales , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos CBA , Vacunas Neumococicas/administración & dosificación , Vacunas Neumococicas/química , Neumonía Neumocócica/inmunología , Neumonía Neumocócica/microbiología , Streptococcus pneumoniae/química , Balance Th1 - Th2RESUMEN
AIM: Detailed characteristic of results of intranasal immunization of mice with one of two variants of vaccinating influenza virus, particularly in combination with a low molecular weight germanium-organic compound (LMW-GOC). An additional aim is evaluation of effect of LMW-GOC on the parameters of immune system in case of intranasal administration of the preparation without the addition of vaccinating virus. MATERIALS AND METHODS: The study was carried out in female CBA mice (18-20 g, 6 animals per group). Intranasal immunization was carried out by 2 different variants of B/Victoria influenza virus--once or twice with a 2 week interval. Cells for study were obtained from spleen and nasal- and bronchial-associated lymphoid tissue (NALT/ BALT) 24 hours and 7 days after intranasal administration of the preparations. The main method of the study--determination of the level of expression of various markers oflymphocytes in comparison with the level of the same markers in the cells of control group animals by using flow cytometry method. The mean parameters obtained were determined by using program package WinMDI 2.8. RESULTS: The main results were the increase of level of expression of various lymphocyte markers obtained from mice after intranasal administration of the vaccines and their combination with LMW-GOC or LMW-GOC only without the participation of the vaccines. A significant increase of the expression of TLR9 marker compared with other parameters was noted. Administration to mice of wild B/Victoria strain notably more frequently conditioned the decrease of expression of some parameters compared with administration of the cold adapted strain. Effect of LMW-GOC without the vaccine also conditioned the increase of levels of markers however a combination of the preparations with the vaccine was more effective. CONCLUSION: The increase of level of expression of a number of lymphocyte markers may serve as a sign of successful intranasal vaccination against influenza. LMW-GOC preparation increases immune stimulating effect of intranasally administered vaccines and in none of the cases weakens the stimulating result of effect of the vaccines, and in many cases increases it. LMW-GOC may be studied as a main or additional adjuvant for intranasal application of influenza vaccines.
Asunto(s)
Adyuvantes Inmunológicos/administración & dosificación , Germanio , Virus de la Influenza B/inmunología , Vacunas contra la Influenza/inmunología , Linfocitos/efectos de los fármacos , Compuestos Organometálicos/administración & dosificación , Infecciones por Orthomyxoviridae/prevención & control , Adyuvantes Inmunológicos/síntesis química , Administración Intranasal , Animales , Anticuerpos Antivirales/sangre , Biomarcadores/metabolismo , Femenino , Expresión Génica , Humanos , Vacunas contra la Influenza/administración & dosificación , Linfocitos/inmunología , Ratones , Ratones Endogámicos CBA , Mucosa Nasal/inmunología , Compuestos Organometálicos/síntesis química , Compuestos Organometálicos/inmunología , Infecciones por Orthomyxoviridae/inmunología , Infecciones por Orthomyxoviridae/virología , Receptor Toll-Like 9/biosíntesis , Receptor Toll-Like 9/inmunología , Vacunación , Vacunas AtenuadasRESUMEN
Fixed erythema--a kind of clinical and histopathologic reaction, fixed drug eruption. The purpose of the study--the study of characteristics of the cytokine profile in patients with erythema and the dynamics of the basal levels of proinflammatory and antiinflammatory cytokines during immunotherapy. All 41 patients with fixed erythema at baseline and after treatment was carried out determination of levels of pro-, anti-inflammatory and regulatory cytokines in the serum by ELISA using test systems "Biosource" (Austria). In patients with fixed erythema Immunovac treatment increased serum IFN-gamma (p < 0.05), IL-1beta (p > 0.05), IL-6. While Kagocel led to an increase in IFN-gamma (p < 0.05), IL-1beta, IL-6 and reduction of TGF-beta (p < 0.05). At the same time in patients with fixed erythema basic therapy contributed to the significant increase in TGF-â and decrease in IL-10. Immunovac-VP-4 had the highest activity for the induction of IFN-gamma. Inclusion in the range of therapeutic and prophylactic measures in patients with fixed erythema immunomodulators promotes activation links innate and adaptive immunity triggers mechanisms, thus increasing the antiviral response in patients with erythema.
Asunto(s)
Antígenos Bacterianos/uso terapéutico , Antivirales/uso terapéutico , Eritema Nudoso/sangre , Eritema Nudoso/terapia , Factores Inmunológicos/uso terapéutico , Inmunoterapia , Inmunidad Adaptativa/efectos de los fármacos , Adulto , Anciano , Antígenos Bacterianos/farmacología , Antivirales/farmacología , Ensayo de Inmunoadsorción Enzimática , Eritema Nudoso/inmunología , Femenino , Humanos , Inmunidad Innata/efectos de los fármacos , Factores Inmunológicos/farmacología , Interferón gamma/sangre , Interferón gamma/inmunología , Interleucina-10/sangre , Interleucina-10/inmunología , Interleucina-1beta/sangre , Interleucina-1beta/inmunología , Interleucina-6/sangre , Interleucina-6/inmunología , Masculino , Persona de Mediana Edad , Factor de Crecimiento Transformador beta/sangre , Factor de Crecimiento Transformador beta/inmunologíaRESUMEN
Cold-adapted (CA) strains A/Krasnodar/35 and B/Victoria/63 were isolated using passages of A/Krasnodar/101/59 and B/Victoria/2/87 wild type strains at low temperatures. The resulting CA strains possessed TS and CA phenotypes and had a reduced ability to reproduce in mouse lungs and nasal turbinates. They displayed a high protective efficacy in experiments on mice. The two CA strains reproduced well in chick embryos and MDCK cell line without change of TS and CA markers. The CA A/Krasnodar/35 strain during passages at low temperature acquired 13 mutations in the 6 internal genes, 8 of those mutations led to amino acid changes. The CA B/Victoria/63 strain acquired 8 mutations in the internal genes, 6 of which led to amino acid changes. The intranasal vaccination of mice with the CA A/Krasnodar/35 strain led to a transitory suppression of various lymphocyte subpopulations, as well as to an increase in the number of some other cell types. The CA strains in question may be used in the future as attenuation donors for live influenza vaccines.
Asunto(s)
Adaptación Fisiológica/genética , Frío , Subtipo H2N2 del Virus de la Influenza A , Vacunas contra la Influenza , Mutación , Sustitución de Aminoácidos , Animales , Línea Celular , Embrión de Pollo , Perros , Humanos , Subtipo H2N2 del Virus de la Influenza A/genética , Subtipo H2N2 del Virus de la Influenza A/inmunología , Subtipo H2N2 del Virus de la Influenza A/metabolismo , Vacunas contra la Influenza/biosíntesis , Vacunas contra la Influenza/genética , Vacunas contra la Influenza/inmunología , Ratones , Vacunas Atenuadas/biosíntesis , Vacunas Atenuadas/genética , Vacunas Atenuadas/inmunologíaRESUMEN
AIM: Detection of features of functioning ofinnate and adaptive immunity pathways in patients with Darier erythema annulare centrifugum (EAC). MATERIALS AND METHODS: 14 EAC patients aged 14 - 52 years were examined. The patients were ranked based on therapy variant. The first group consisted of 6 patients who had received Immunovac-VP-4 (Immunovac) against the background of basic therapy; the second group (4 patients) received cagocel against the background of basic therapy; the third group (4 patients) received only basic therapy; the group of healthy individuals consisted of 15 individuals. All the patients had the level ofcytokines in blood sera determined by solid-phase EIA by using Biosource (Austria) test-systems. Evaluation of TLR expression in peripheral blood mononuclear lymphocytes and keratinocytes was carried out by flow cytometry method by using monoclonal antibodies (Catlag Laboratories, USA) against the corresponding antigens; evaluation of content of lymphocyte subpopulations in blood was carried out by using monoclonal antibodies by flow cytometry method on FacsCalibur flow cytometer (Becton Dickinson, USA). RESULTS: Immunotherapy by Immunovac and cagocel facilitated the increase of CD3+, CD4+, CD8+. Immunovac facilitated a significant increase ofinitially low values ofCD25+, CD95+ and normalization of CD72+; normalization of IgM level. Immunovac increased the level of serum IL-2, induced the increase of IFN-gamma synthesis in contrast to cagocel and basic therapy, the administration of those caused its decrease. TGF-beta increased during the course of Immunovac immunotherapy and decreased during basic therapy. In the course of basic therapy a significant increase of initially high level ofcytokine IL-I beta was observed. CONCLUSION: Immunovac therapy resulted in correction of content of lymphocyte populations, sera cytokines, facilitating the normalization of immunocompetent cell proliferation processes, activation of NK-cells, macrophages and at the same time suppression of DTH reactions. Immunovac facilitated the enhancement ofTLR3, 9 expression in the skin that indicates the inclusion ofintracellular receptor mechanisms of innate immunity.