RESUMEN
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.
Asunto(s)
Artritis Experimental/metabolismo , Artritis Experimental/terapia , Artritis Reumatoide/metabolismo , Artritis Reumatoide/terapia , Proteínas Portadoras/administración & dosificación , Proteínas Portadoras/genética , Terapia Genética/métodos , Proteínas Virales/administración & dosificación , Proteínas Virales/genética , Animales , Artritis Experimental/patología , Artritis Reumatoide/patología , Femenino , Vectores Genéticos , Miembro Posterior/patología , Inyecciones Intramusculares , Masculino , Ratas Wistar , Sinovitis/metabolismo , Sinovitis/patología , Sinovitis/terapia , Resultado del Tratamiento , Virus de la ViruelaRESUMEN
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.
Asunto(s)
Macrófagos/virología , Viruela/prevención & control , Virus de la Viruela/fisiología , Virión/crecimiento & desarrollo , Adulto , Animales , Anticuerpos Antivirales/sangre , Granulocitos/inmunología , Humanos , Macrófagos/ultraestructura , Masculino , Microscopía Electrónica , Especificidad de Órganos , Cultivo Primario de Células , Viruela/sangre , Viruela/inmunología , Viruela/virología , Vacuna contra Viruela/farmacología , Virus de la Viruela/ultraestructura , Virión/ultraestructura , Replicación ViralRESUMEN
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.
Asunto(s)
Alquenos/farmacología , Antivirales/farmacología , Benzamidas/farmacología , Hidrazinas/farmacología , Isoindoles/farmacología , Viruela/tratamiento farmacológico , Virus de la Viruela/efectos de los fármacos , Administración Intranasal , Animales , Modelos Animales de Enfermedad , Células Epiteliales/efectos de los fármacos , Células Epiteliales/patología , Células Epiteliales/virología , Humanos , Pulmón/efectos de los fármacos , Pulmón/patología , Pulmón/virología , Macrófagos Alveolares/efectos de los fármacos , Macrófagos Alveolares/patología , Macrófagos Alveolares/virología , Ratones , Ratones Endogámicos ICR , Viruela/patología , Viruela/virología , Virus de la Viruela/fisiología , Carga Viral/efectos de los fármacos , Replicación Viral/efectos de los fármacosRESUMEN
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.
RESUMEN
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.
Asunto(s)
Anticuerpos Antivirales/sangre , Inmunización , Interleucina-2/genética , Interleucina-2/inmunología , Infecciones por Poxviridae/sangre , Infecciones por Poxviridae/inmunología , Vacuna contra Viruela/inmunología , Virus Vaccinia/genética , Virus Vaccinia/inmunología , Animales , Línea Celular , Citocinas/sangre , Humanos , Técnicas para Inmunoenzimas , Inyecciones Subcutáneas , Interleucina-2/sangre , Activación de Linfocitos , Masculino , Ratones , Ratones Endogámicos BALB C , Vacuna contra Viruela/administración & dosificación , Vacuna contra Viruela/genética , Bazo/inmunología , Células TH1/inmunología , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/inmunologíaAsunto(s)
Vacunas contra el SIDA/inmunología , Vacunas contra Hepatitis B/inmunología , Inmunidad Mucosa , Plantas Modificadas Genéticamente/inmunología , Solanum lycopersicum/inmunología , Vacunas contra el SIDA/genética , Animales , Anticuerpos Antivirales/inmunología , VIH-1/genética , VIH-1/inmunología , Vacunas contra Hepatitis B/genética , Virus de la Hepatitis B/genética , Virus de la Hepatitis B/inmunología , Inmunoglobulina A/inmunología , Solanum lycopersicum/genética , Ratones , Ratones Endogámicos BALB C , Plantas Modificadas Genéticamente/genéticaRESUMEN
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.
Asunto(s)
Vacunas contra el SIDA , Vacunas contra Hepatitis B , Plantas Modificadas Genéticamente , Solanum lycopersicum , Vacunas contra el SIDA/inmunología , Animales , Epítopos , Productos del Gen env/genética , Productos del Gen env/inmunología , Productos del Gen gag/genética , Productos del Gen gag/inmunología , VIH-1/inmunología , Hepatitis B/inmunología , Antígenos de Superficie de la Hepatitis B/genética , Antígenos de Superficie de la Hepatitis B/inmunología , Vacunas contra Hepatitis B/inmunología , Solanum lycopersicum/genética , Solanum lycopersicum/inmunología , Solanum lycopersicum/metabolismo , Ratones , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/inmunología , Plantas Modificadas Genéticamente/metabolismo , Proteínas Recombinantes de Fusión/biosíntesis , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/inmunología , Vacunas Comestibles/inmunologíaAsunto(s)
Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Orthopoxvirus/patogenicidad , Receptores del Factor de Necrosis Tumoral/inmunología , Receptores del Factor de Necrosis Tumoral/metabolismo , Factor de Necrosis Tumoral alfa/farmacología , Secuencia de Aminoácidos , Animales , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/inmunología , Células Cultivadas , Escherichia coli/genética , Escherichia coli/metabolismo , Fibroblastos/inmunología , Regulación Viral de la Expresión Génica , Genes Virales , Humanos , Ratones , Datos de Secuencia Molecular , Orthopoxvirus/metabolismo , Proteínas Recombinantes/inmunología , Proteínas Recombinantes/metabolismo , Alineación de Secuencia/métodos , Especificidad de la Especie , Transfección/métodos , Factor de Necrosis Tumoral alfa/inmunología , Factor de Necrosis Tumoral alfa/metabolismoAsunto(s)
VIH/inmunología , Hepatitis B/inmunología , Orthopoxvirus/inmunología , Ingeniería de Proteínas/métodos , Vacunas Virales/biosíntesis , Vacunas Virales/inmunología , Vacunas contra el SIDA , Animales , Diseño de Fármacos , Ingeniería Genética/métodos , Humanos , Ratones , Proteínas Recombinantes/inmunología , Vacunas Atenuadas/biosíntesis , Vacunas Atenuadas/clasificación , Vacunas Atenuadas/inmunología , Vacunas Sintéticas , Vacunas contra Hepatitis Viral , Vacunas Virales/clasificaciónRESUMEN
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).
Asunto(s)
Hepacivirus/genética , Hepatitis C/epidemiología , Personal de Hospital , Biomarcadores , Infección Hospitalaria/epidemiología , Infección Hospitalaria/virología , Estudios Transversales , Femenino , Hepacivirus/aislamiento & purificación , Hepatitis C/genética , Hepatitis C/microbiología , Hepatitis C/transmisión , Hospitales de Distrito , Humanos , Masculino , Factores de Riesgo , Federación de Rusia/epidemiologíaRESUMEN
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.
Asunto(s)
Interferón-alfa/uso terapéutico , Enfermedad del Virus de Marburg/prevención & control , Enfermedad Aguda , Animales , Sangre/virología , Temperatura Corporal , Chlorocebus aethiops , Cobayas , Humanos , Interferón alfa-2 , Interferón-alfa/inmunología , Enfermedad del Virus de Marburg/inmunología , Marburgvirus/fisiología , Proteínas Recombinantes , Sobrevida , ViremiaAsunto(s)
Antígenos CD/genética , Receptores de Interferón/genética , Virus de la Viruela/genética , Proteínas Virales/genética , Secuencia de Aminoácidos , Secuencia de Bases , Clonación Molecular , ADN Recombinante , Escherichia coli/genética , Humanos , Datos de Secuencia Molecular , Plásmidos , Receptor de Interferón gammaRESUMEN
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.
Asunto(s)
Interferón gamma/antagonistas & inhibidores , Receptores de Interferón/metabolismo , Virus de la Viruela/patogenicidad , Proteínas Estructurales Virales/farmacología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Escherichia coli/genética , Genes Virales/genética , Humanos , Ratones , Datos de Secuencia Molecular , Receptores de Interferón/química , Receptores de Interferón/genética , Receptores de Interferón/aislamiento & purificación , Proteínas Recombinantes de Fusión/biosíntesis , Proteínas Recombinantes de Fusión/aislamiento & purificación , Proteínas Recombinantes de Fusión/farmacología , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Virus de la Viruela/genética , Proteínas Estructurales Virales/genética , Proteínas Estructurales Virales/aislamiento & purificación , VirulenciaAsunto(s)
Interferón gamma/fisiología , Virus de la Viruela/genética , Proteínas Virales/fisiología , Secuencia de Aminoácidos , Escherichia coli/genética , Humanos , Interferón gamma/inmunología , Datos de Secuencia Molecular , Receptores de Interferón/genética , Proteínas Recombinantes/genética , Proteínas Virales/genética , Receptor de Interferón gammaRESUMEN
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.