RESUMO
Wistar rats with collagen-induced arthritis were intramuscularly injected with the recombinant plasmid pcDNA/sTNF-BD encoding the sequence of the TNF-binding protein domain of variola virus CrmB protein (VARV sTNF-BD) or the pcDNA3.1 vector. Quantitative analysis showed that the histopathological changes in the hind-limb joints of rats were most severe in the animals injected with pcDNA3.1 and much less severe in the group of rats injected with pcDNA/sTNF-BD, which indicates that gene therapy of rheumatoid arthritis is promising in the case of local administration of plasmids governing the synthesis of VARV immunomodulatory proteins.
Assuntos
Artrite Experimental/metabolismo , Artrite Experimental/terapia , Artrite Reumatoide/metabolismo , Artrite Reumatoide/terapia , Proteínas de Transporte/administração & dosagem , Proteínas de Transporte/genética , Terapia Genética/métodos , Proteínas Virais/administração & dosagem , Proteínas Virais/genética , Animais , Artrite Experimental/patologia , Artrite Reumatoide/patologia , Feminino , Vetores Genéticos , Membro Posterior/patologia , Injeções Intramusculares , Masculino , Ratos Wistar , Sinovite/metabolismo , Sinovite/patologia , Sinovite/terapia , Resultado do Tratamento , Vírus da VaríolaRESUMO
Studies of the primary cultures of granulocytes, mononuclear, and monocyte-macrophage cells derived from human blood were performed using variola virus (VARV) in the doses of 0.001-0.021 PFU/cell (plaques-forming units per cell). Positive dynamics of the virus accumulation was observed only in the monocyte-macrophages with maximum values of virus concentration (5.0-5.5 Ig PFU/ml) mainly within six days after the infection. The fact of VARV replication in the monocyte-macrophages was confirmed by the data of electron microscopy. At the same time, virus vaccines when tested in doses 3.3 and 4.2 Ig PFU/ml did not show the ability to reproduce in these human cells. The people sensitivity to VARV as assessed from the data obtained on human monocyte-macrophages corresponded to -1 PFU (taking into account the smooth interaction of the virus in the body to the cells of this type), which is consistent to previously found theoretical data on the virus sensitivity. The human susceptibility to VARV assessed experimentally can be used to predict the adequacy of developed smallpox models (in vivo) based on susceptible animals. This is necessary for reliable assessment of the efficiency of development of drugs for treatment and prophylaxis of the smallpox.
Assuntos
Macrófagos/virologia , Varíola/prevenção & controle , Vírus da Varíola/fisiologia , Vírion/crescimento & desenvolvimento , Adulto , Animais , Anticorpos Antivirais/sangue , Granulócitos/imunologia , Humanos , Macrófagos/ultraestrutura , Masculino , Microscopia Eletrônica , Especificidade de Órgãos , Cultura Primária de Células , Varíola/sangue , Varíola/imunologia , Varíola/virologia , Vacina Antivariólica/farmacologia , Vírus da Varíola/ultraestrutura , Vírion/ultraestrutura , Replicação ViralRESUMO
Mice of the ICR outbred population were infected intranasally (i/n) with the variola virus (VARV, strain Ind-3a). Clinical signs of the disease did not appear even at the maximum possible dose of the virus 5.2 lg PFU/head (plaque-forming units per head). In this case, 50% infective dose (ID50) of VARV estimated by the presence or absence of the virus in the lungs three days after infection (p.i.) was equal to 2.7 ± 0.4 lg PFU/head. Taking into account the 10% application of the virus in the lungs during the intranasal infection of the mice, it was adequate to 1.7 lg PFU/lungs. This indicates a high infectivity of the VARV for mice comparable to its infectivity for humans. After the i/n infection of mice with the VARV at a dose 30 ID50/ head the highest concentration of the virus detected in the lungs (4.9 ± 0.0 lg PFU/ml of homogenate) and in nasal cavity tissues (4.8 ± 0.0 lg PFU/ml) were observed. The pathomorphological changes in the respiratory organs of the mice infected with the VARV appeared at 3-5 days p.i., and the VARV reproduction noted in the epithelial cells and macrophages were noticed. When the preparations ST-246 and NIOCH-14 were administered orally at a dose of 60 µg/g of mouse weight up to one day before infection, after 2 hours, 1 and 2 days p.i., the VARV reproduction in the lungs after 3 days p.i. decreased by an order of magnitude. Thus, outbred ICR mice infected with the VARV can be used as a laboratory model of the smallpox when evaluating the therapeutic and prophylactic efficacy of the antismallpox drugs.
Assuntos
Alcenos/farmacologia , Antivirais/farmacologia , Benzamidas/farmacologia , Hidrazinas/farmacologia , Isoindóis/farmacologia , Varíola/tratamento farmacológico , Vírus da Varíola/efeitos dos fármacos , Administração Intranasal , Animais , Modelos Animais de Doenças , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/patologia , Células Epiteliais/virologia , Humanos , Pulmão/efeitos dos fármacos , Pulmão/patologia , Pulmão/virologia , Macrófagos Alveolares/efeitos dos fármacos , Macrófagos Alveolares/patologia , Macrófagos Alveolares/virologia , Camundongos , Camundongos Endogâmicos ICR , Varíola/patologia , Varíola/virologia , Vírus da Varíola/fisiologia , Carga Viral/efeitos dos fármacos , Replicação Viral/efeitos dos fármacosRESUMO
We propose a model of rheumatoid arthritis (RA) induced in outbred guinea pigs using a single subcutaneous injection of complete Freund's adjuvant to the hind paw. Histological examination of this model shows fibrin deposition on the surface of the synovial membrane, leukocyte infiltration of the synovial membrane and adjacent tissues, proliferation of the granulation tissue, and emergence of angioid areas, characteristic of RA. The cell response appears as an increase in the plasma cell count and development of follicle-like lymphoid infiltrates; erosion of the articular surface of the cartilage, frequently with deep cartilage destruction over large areas; and epiphysiopathy. The high reproducibility of arthritis induction in this RA model has been demonstrated. The proposed model is promising for the assessment of anti-arthritis preparations and dosage regimens.
RESUMO
A genetic construct of the human interleukin-2 (IL-2) gene within vaccinia virus (L-IVP strain) has been designed. The authors show the capacity of CV-1 cells infected with the recombinant vaccinia virus VV-SIL2 to secrete human IL-2 into the culture medium. Human IL-2 has been detected by immunoblotting. The sera from the animals immunized with the recombinant virus VV-SIL2 exhibited both human IL-2 and its antibodies throughout the observation period. This recombinant virus immunization induced both humoral and cell-mediated immune responses to human IL-2; the observed changes in the concentrations of cytokines are likely to suggest that the response predominantly followed a Th1 pathway. The study construct was nontoxic at the used concentrations and administration routes. The findings point that it is promising to investigate the adjuvant properties of the recombinant VV-SIL2 vaccine-based preparation for immunization in combination with various vaccines and to study this construct in therapy for cancer diseases.
Assuntos
Anticorpos Antivirais/sangue , Imunização , Interleucina-2/genética , Interleucina-2/imunologia , Infecções por Poxviridae/sangue , Infecções por Poxviridae/imunologia , Vacina Antivariólica/imunologia , Vaccinia virus/genética , Vaccinia virus/imunologia , Animais , Linhagem Celular , Citocinas/sangue , Humanos , Técnicas Imunoenzimáticas , Injeções Subcutâneas , Interleucina-2/sangue , Ativação Linfocitária , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Vacina Antivariólica/administração & dosagem , Vacina Antivariólica/genética , Baço/imunologia , Células Th1/imunologia , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/imunologiaAssuntos
Vacinas contra a AIDS/imunologia , Vacinas contra Hepatite B/imunologia , Imunidade nas Mucosas , Plantas Geneticamente Modificadas/imunologia , Solanum lycopersicum/imunologia , Vacinas contra a AIDS/genética , Animais , Anticorpos Antivirais/imunologia , HIV-1/genética , HIV-1/imunologia , Vacinas contra Hepatite B/genética , Vírus da Hepatite B/genética , Vírus da Hepatite B/imunologia , Imunoglobulina A/imunologia , Solanum lycopersicum/genética , Camundongos , Camundongos Endogâmicos BALB C , Plantas Geneticamente Modificadas/genéticaRESUMO
The synthetic chimeric gene TBI-HBS encoding the synthesis of immunogenic ENV and GAC epitopes of HIV-1 (immunogenes of T- and B-lymphocytes) and of the surface protein (HBsAg) of the hepatitis B virus was introduced into tomato plants var. Ventura by agrobacterial vector pBIN35TBI-HBS; transgenic tomato plants with the integrated gene TBI-HBS were generated. The integration of the TBI-HBS target gene was confirmed by PCR. The synthesis of antigenic proteins of TBI and HBsAg in fruits of transgenic tomato plants was displayed by immunoassay. The fruits of transgenic tomato plants were fed to experimental mice with a 1-week interval. On days 14 and 28, there was discovered a sufficiently high content of antibodies to the antigenic proteins of HBV and HIV-1 in serum of experimental animals. Antibodies were found in feces of experimental mice; no antibodies were found in the control group of mice. Hence, it was established that the TBI (HIV-1) and HBsAg (HBV) antigens were synthesized in transgenic tomato fruits due to the integrated construction of pBINNp35TBI-HBS in an amount that was enough to induce the immunogenic response in mice to the oral delivery of edible vaccine.
Assuntos
Vacinas contra a AIDS , Vacinas contra Hepatite B , Plantas Geneticamente Modificadas , Solanum lycopersicum , Vacinas contra a AIDS/imunologia , Animais , Epitopos , Produtos do Gene env/genética , Produtos do Gene env/imunologia , Produtos do Gene gag/genética , Produtos do Gene gag/imunologia , HIV-1/imunologia , Hepatite B/imunologia , Antígenos de Superfície da Hepatite B/genética , Antígenos de Superfície da Hepatite B/imunologia , Vacinas contra Hepatite B/imunologia , Solanum lycopersicum/genética , Solanum lycopersicum/imunologia , Solanum lycopersicum/metabolismo , Camundongos , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/imunologia , Plantas Geneticamente Modificadas/metabolismo , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Vacinas de Plantas Comestíveis/imunologiaAssuntos
Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Orthopoxvirus/patogenicidade , Receptores do Fator de Necrose Tumoral/imunologia , Receptores do Fator de Necrose Tumoral/metabolismo , Fator de Necrose Tumoral alfa/farmacologia , Sequência de Aminoácidos , Animais , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/imunologia , Células Cultivadas , Escherichia coli/genética , Escherichia coli/metabolismo , Fibroblastos/imunologia , Regulação Viral da Expressão Gênica , Genes Virais , Humanos , Camundongos , Dados de Sequência Molecular , Orthopoxvirus/metabolismo , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/metabolismo , Alinhamento de Sequência/métodos , Especificidade da Espécie , Transfecção/métodos , Fator de Necrose Tumoral alfa/imunologia , Fator de Necrose Tumoral alfa/metabolismoAssuntos
HIV/imunologia , Hepatite B/imunologia , Orthopoxvirus/imunologia , Engenharia de Proteínas/métodos , Vacinas Virais/biossíntese , Vacinas Virais/imunologia , Vacinas contra a AIDS , Animais , Desenho de Fármacos , Engenharia Genética/métodos , Humanos , Camundongos , Proteínas Recombinantes/imunologia , Vacinas Atenuadas/biossíntese , Vacinas Atenuadas/classificação , Vacinas Atenuadas/imunologia , Vacinas Sintéticas , Vacinas contra Hepatite Viral , Vacinas Virais/classificaçãoRESUMO
The occurrence of markers, the genotypic variety of isolates and the profile of risk factors with respect to viral hepatitis C among 629 employees of the Regional Clinical Hospital (RCH) in Novosibirsk and 1,020 employees of the Central District Hospital (CDH) in Iskitim were studied in a cross-sectional investigation. The occurrence of hepatitis C virus (HCV) markers was 5.1% in RCH and 2.2% in CDH. Among the risk factors in the population under study were: the medical history of blood transfusions (TF) with 0 TF, anti-HCV = 2.3%; 1 TF, = 5.7% > 1 TF, = 13.5% (p < 0.001); general anesthesia (GA) with < or = 2 GA, anti-HCV = 2.8%; > 2 GA, = 7.8% (p = 0.002); surgical interventions (SU) with 0 SU, = 1.9%; > 0 SU, = 4.3% (p = 0.012); the intravenous use of drugs (OR = 31.8); age (< or = 25 years, anti-HCV IgG = 8.6% > 25 years, = 4.5%); the number of partners of the opposite sex < or = 4 partners, = 2.4%; > 4 partners, = 6.9%; p < 0.001). The probable risk factors at a working place (pricks and cuts, contamination of mucous membranes with blood and other biological fluids, etc.) proved to be faintly related with the status of HBV infection. HBV isolates detected in the examined persons (35 examinees) were distributed by genotypes as follows: 60% of subtype 1b, 28.6% of subtype 2a/2c, 11.4% of subtype 3a. HBV of genotype 1a was not detected in the examined specimens, while the detection rate of genotype 2a/2c was considerably greater than in specimens obtained in the European and Asian parts of Russia (according to the data reported earlier).
Assuntos
Hepacivirus/genética , Hepatite C/epidemiologia , Recursos Humanos em Hospital , Biomarcadores , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/virologia , Estudos Transversais , Feminino , Hepacivirus/isolamento & purificação , Hepatite C/genética , Hepatite C/microbiologia , Hepatite C/transmissão , Hospitais de Distrito , Humanos , Masculino , Fatores de Risco , Federação Russa/epidemiologiaRESUMO
The possibility of emergency prophylaxis of Marburg hemorrhagic fever with leukocytic and recombinant interferons was studied in experiments on Cercopithecus aethiops. None of the agents protected monkeys from the action of lethal doses of Marburg virus. Recombinant interferon-alpha(2)administered according to the emergency prophylaxis schedule prolonged the mean life-span of monkeys injected with Marburg virus in doses of 100 and 1000 LD50 by 1.9 and 6.1 days, respectively.
Assuntos
Interferon-alfa/uso terapêutico , Doença do Vírus de Marburg/prevenção & controle , Doença Aguda , Animais , Sangue/virologia , Temperatura Corporal , Chlorocebus aethiops , Cobaias , Humanos , Interferon alfa-2 , Interferon-alfa/imunologia , Doença do Vírus de Marburg/imunologia , Marburgvirus/fisiologia , Proteínas Recombinantes , Sobrevida , ViremiaAssuntos
Antígenos CD/genética , Receptores de Interferon/genética , Vírus da Varíola/genética , Proteínas Virais/genética , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , DNA Recombinante , Escherichia coli/genética , Humanos , Dados de Sequência Molecular , Plasmídeos , Receptor de Interferon gamaRESUMO
To study specific properties of the human gamma-interferon (gamma-IFN) receptor-like proteins of the highly virulent and low virulent strains of variola (smallpox) virus (VAR) recombinant plasmids determining synthesis of these proteins in E. coli cells have been constructed. The recombinant viral gamma-IFN receptor-like proteins have been found to have high interferon-neutralising activity with regard to human gamma-IFN but not murine gamma-IFN and human alpha-IFN. The variola major and variola minor proteins under study do not differ in the efficiency of human gamma-IFN antiviral activity inhibition.
Assuntos
Interferon gama/antagonistas & inibidores , Receptores de Interferon/metabolismo , Vírus da Varíola/patogenicidade , Proteínas Estruturais Virais/farmacologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Escherichia coli/genética , Genes Virais/genética , Humanos , Camundongos , Dados de Sequência Molecular , Receptores de Interferon/química , Receptores de Interferon/genética , Receptores de Interferon/isolamento & purificação , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/isolamento & purificação , Proteínas Recombinantes de Fusão/farmacologia , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Vírus da Varíola/genética , Proteínas Estruturais Virais/genética , Proteínas Estruturais Virais/isolamento & purificação , VirulênciaAssuntos
Interferon gama/fisiologia , Vírus da Varíola/genética , Proteínas Virais/fisiologia , Sequência de Aminoácidos , Escherichia coli/genética , Humanos , Interferon gama/imunologia , Dados de Sequência Molecular , Receptores de Interferon/genética , Proteínas Recombinantes/genética , Proteínas Virais/genética , Receptor de Interferon gamaRESUMO
The authors have obtained several variants of liposomal forms of human alpha 2-interferon. The forms intended to preserve antiviral activity with regard to the methods of liposomal formation were comparatively studied. It has been found that liposomal formation through stirring caused no decrease in antiviral activity.