RESUMEN
AIM: To study the effect of chimeric E7 protein of human papillomavirus type 18 on activation of adaptive immunity in absence of adjuvant. MATERIALS AND METHODS: Chimeric protein was genetically engineered and represents the protein molecule consisting of full-size E7 oncoprotein and heat-shock protein 70 (HSP70) of Mycobacterium tuberculosis in one polypeptide chain. Antibody titers as well as isotypes and subisotypes of immunoglobulins were measured by ELISA in sera of immunized animals. RESULTS: It was shown that studied construction E7 (HPV-18)-HSP70 significantly increases titers of antibodies to E7 protein of HPV type 18 and have cross-reactive antigenic activity with E7 protein of HPV type 16. Immunization with chimeric protein resulted in increase of IgG1 and IgG2b levels and decrease of IgG2a and IgM levels. CONCLUSION: . Oncoprotein E7 included in chimeric construction with HSP70 could be used for further studies on development of therapeutic vaccine for treatment of cervical cancer and precancerous lesions. Skew of immune response to Th2 type after intraperitoneal administration of the studied construction points to necessity for control of immunity during such studies.
Asunto(s)
Proteínas Bacterianas/inmunología , Proteínas de Unión al ADN/inmunología , Proteínas HSP70 de Choque Térmico/inmunología , Proteínas Oncogénicas Virales/inmunología , Infecciones por Papillomavirus/prevención & control , Vacunas contra Papillomavirus/inmunología , Proteínas Recombinantes de Fusión/inmunología , Neoplasias del Cuello Uterino/prevención & control , Adyuvantes Inmunológicos/administración & dosificación , Animales , Anticuerpos Antivirales/sangre , Especificidad de Anticuerpos , Proteínas Bacterianas/administración & dosificación , Proteínas Bacterianas/genética , Proteínas de Unión al ADN/administración & dosificación , Proteínas de Unión al ADN/genética , Evaluación Preclínica de Medicamentos , Femenino , Proteínas HSP70 de Choque Térmico/administración & dosificación , Proteínas HSP70 de Choque Térmico/genética , Humanos , Inmunización , Esquemas de Inmunización , Inmunoglobulina A/sangre , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Inyecciones Intraperitoneales , Masculino , Ratones , Proteínas Oncogénicas Virales/administración & dosificación , Proteínas Oncogénicas Virales/genética , Infecciones por Papillomavirus/sangre , Vacunas contra Papillomavirus/administración & dosificación , Vacunas contra Papillomavirus/genética , Proteínas Recombinantes de Fusión/administración & dosificación , Proteínas Recombinantes de Fusión/genética , Neoplasias del Cuello Uterino/inmunología , Neoplasias del Cuello Uterino/virologíaRESUMEN
A new reagent for a skin test given the name Diaskintest has been designed for the screening diagnosis of tuberculosis and preclinical and clinical trials conducted. Preclinical trials were carried out on 315 laboratory animals (guinea-pigs, albino mice). The reagent Diaskintest was ascertained to be nontoxic, to have no sensitizing properties, to be safe and specific, and to induce no positive reactions in BCG-vaccinated animals and healthy guinea-pigs. Its specific activity was comparable with that of the national reference--purified tuberculin PPD-L-2. With progression of tuberculous lesions, the guinea-pigs showed higher responses to Diaskintest dilution and the BCG-vaccinated animals lacked responses to Diaskintest with increased delayed type hypersensitivity. The clinical trial was permitted by the Federal Service for Surveillance in Health Care and Social Development of the Russian Federation. Clinical trials were conducted in 150 persons. The safety, specificity, sensitivity of Diaskintest were first examined in the clinical studies and its action was compared with the results of tuberculin skin test (Mantoux test) with 2 TE of PPD L-2. Diaskintest was ascertained to be highly sensitive when given in a dose of 0.2 microg in 0.1 ml. In patients with active tuberculosis and new cases of Mycobacterium tuberculosis infection, the agent induced a positive skin reaction (a papule of more than 10 mm) in 98-100% of cases (p < 0.05). The agent caused no reaction associated with BCG vaccination. The specificity of the test was 93-100% with 95% significance. The rate of overexuberant reactions (vesicular necrotic changes, lymphangitis, and lymphadenitis) was 4-14% with 95% significance. Tuberculosis patients with significant immunopathological disorders might have no skin sensitivity to Diaskintest, as to PPD L-2 (a negative test). The findings substantiate the use of Diaskintest for mass epidemiological surveys for the differential diagnosis of tuberculosis and BCG vaccination-associated complications. The agent may be also used to evaluate the activity of the process in patients with tuberculosis and the efficiency of treatment in combination with other methods and to make a differential diagnosis of tuberculosis.
Asunto(s)
Prueba de Tuberculina/métodos , Tuberculosis/diagnóstico , Adolescente , Adulto , Animales , Niño , Diagnóstico Diferencial , Modelos Animales de Enfermedad , Cobayas , Humanos , Ratones , Persona de Mediana Edad , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Adulto JovenRESUMEN
The fact that antibiotics exert no effect on Mycobacterium tuberculosis may be attributable to the production of their persistent forms. A correlation was studied between the expression of the genes Rv3286c, Rv2626c, Rv2031, and Rv3133c and the phenotypical tolerance to antibiotics in relation to the physiological condition of M. bovis BCG. The anaerobic growth conditions ("Wayne dormancy model") were found to be attended by the increased expression of all the studied genes, by the higher tolerance to rifampicin, and by the decreased phenotypical resistance to metronidazole.
Asunto(s)
Antibióticos Antituberculosos/farmacología , Farmacorresistencia Bacteriana/genética , Expresión Génica/fisiología , Genes Bacterianos/genética , Metronidazol/farmacología , Mycobacterium bovis/genética , Rifampin/farmacología , Animales , Recuento de Colonia Microbiana , ADN Bacteriano/genética , Expresión Génica/efectos de los fármacos , Genes Bacterianos/efectos de los fármacos , Técnicas In Vitro , Mycobacterium bovis/efectos de los fármacos , Mycobacterium bovis/crecimiento & desarrollo , Fenotipo , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
The type of human papilloma virus (HPV) was determined in 26 children aged between 1 year 10 months to 15 years 5 months suffering from recurrent respiratory papillomatosis (RRP). Polymerase chain reaction identified DNA of HPV type 6, 11, 16 and 18. HPV DNA was detected in all the patients including fifteen patients infected with HPV type 11; seven patients infected with HPV type 6; four children infected with HPV type 6 and 11. Types 16 and 18 of HPV were not detected. The analysis of RRP course has found that laryngeal papillomatosis runs a more aggressive course in cases with HPV type 11 infection than in those with HPV type 6.
Asunto(s)
Neoplasias Laríngeas/patología , Neoplasias Laríngeas/virología , Papiloma/patología , Papiloma/virología , Papillomaviridae/aislamiento & purificación , Adolescente , Niño , Preescolar , ADN Viral , Femenino , Humanos , Lactante , Neoplasias Laríngeas/epidemiología , Masculino , Papiloma/epidemiología , Papillomaviridae/clasificación , Papillomaviridae/genética , Reacción en Cadena de la Polimerasa , Prevalencia , RecurrenciaRESUMEN
Epizootological, serological, and molecular virological analysis of an outbreak of hemorrhagic fever with renal syndrome (HFRS) in the Egoryevsk district of Moscow region (September 1995-January 1996) has been carried out. Hantavirus (Puumala) antigen and virus-specific antibodies were isolated from bank voles captured in the endemic focus. Anti-Puumala antibodies were detected in the sera of all HFRS patients and in 2% healthy residents of the endemic focus. Analysis of nucleotide sequence (RNA from hantavirus-positive lung of a bank vole) showed that the studied hantavirus is a distinct genotype of Puumala virus. Hence, a new highly active natural focus of HFRS associated with Puumala virus, dangerous for the population, has been revealed in Moscow region.
Asunto(s)
Brotes de Enfermedades , Fiebre Hemorrágica con Síndrome Renal/epidemiología , Animales , Reservorios de Enfermedades , Virus Hantaan , Humanos , Moscú/epidemiologíaRESUMEN
Enzyme immunoassay with biotin-labeled monoclonal antibodies to Puumala virus was used for typing hantavirus antigens. The system is type-specific, that is, it detects Puumala virus only, which was demonstrated by typing 9 hantavirus antigens in infected Vero E5 cells and by epizootological analysis of an HFRS outbreak in the Yegoryevsk district of the Moscow region in November-December 1995. The suggested system may be used as an element of EIA screening of natural samples in analysis of the screening results.
Asunto(s)
Antígenos Virales/análisis , Orthohantavirus/inmunología , Anticuerpos Monoclonales , Biotina , Técnicas para InmunoenzimasRESUMEN
The possibility of the atypical course of tularemia with the prolonged persistence of Francisella tularensis in common voles (M. arvalis), the twin species of East European voles (M. rossiaemeridionalis), was studied. Experiments were made on 33 animals grown in the laboratory. F. tularensis strain 165 was used. The animals were infected by feeding them according to the previously developed scheme. 7 out of 33 voles showed the atypical course of tularemia: in 3 voles the disease took a prolonged course with bacteriuria and death on days 25-34; 3 other voles with bacteriuria registered before days 33, 66 and 172 (the term of observation) survived. The surviving animals were killed on day 183, and the presence of bacteria in their organs and seroconversion were established. One vole excreted no bacteria with urine and had no bacteria in its organs (the animal was examined on day 156), but in its blood specific antibodies were detected. To determine bacteriuria, the immunofluorescence test was used together with biological assays. Thus, M. arvalis, like M. rossiaemeridionalis studied earlier, can harbor F. tularensis at the period between epizootics. When voles of the former species penetrate stacks of straw and hayricks, conditions appear for the transfer of the infection to the latter species, M. rossiaemeridionalis. Therefore, in the foci of the meadow-field type each of these two species of voles may be not only of epizootic, but also of epidemic importance.
Asunto(s)
Arvicolinae , Tularemia/veterinaria , Animales , Animales de Laboratorio , Anticuerpos Antibacterianos/análisis , Especificidad de Anticuerpos , Bacteriuria/inmunología , Bacteriuria/microbiología , Bacteriuria/veterinaria , Modelos Animales de Enfermedad , Francisella tularensis/inmunología , Francisella tularensis/aislamiento & purificación , Francisella tularensis/patogenicidad , Factores de Tiempo , Tularemia/inmunología , Tularemia/microbiología , VirulenciaAsunto(s)
Garrapatas/fisiología , Animales , Ecología , Femenino , Masculino , Densidad de Población , Estaciones del Año , Razón de Masculinidad , SiberiaRESUMEN
The autumn-winter (1977-1978) tularemia epizootic in small murine rodents was revealed and studied at the natural focus of the meadow and field type in the south of the Moscow Region. The efficacy of the serologic method (the antibody neutralization test) of studying the organs of the caught rodents and the bodies of dead rodents was found to be greater than that of the traditional bacteriologic methods (26.6% and 9.6%, respectively). The serologic study of 908 specimens of avian excrements collected during the period from autumn to spring (1977-1978) revealed that tularemia antigen could be constantly detected, starting from October. The serologic method was effective when used both for the early and retrospective detection of the infective agent and allowed to characterize the epizootic process in greater detail.