RESUMEN
L-lysin-a-oxidase (LO), a fungal enzyme catalysing oxidative deamination of L-lysin, was used for the inhibition of the reproduction of herpes simplex virus type 1 (HSV-1). Antiviral activity of LO was tested in vitro. The expression of viral antigens and CPE of HSV-1 was inhibited by LO at a concentration 0.7 mg/ml.
Asunto(s)
Aminoácido Oxidorreductasas/farmacología , Antivirales/farmacología , Herpesvirus Humano 1/efectos de los fármacos , Replicación Viral/efectos de los fármacos , Animales , Antígenos Virales/efectos de los fármacos , Chlorocebus aethiops , Efecto Citopatogénico Viral/efectos de los fármacos , Herpesvirus Humano 1/inmunología , Herpesvirus Humano 1/fisiología , Células VeroRESUMEN
The authors prepared 156 mouse hybridomas producing monoclonal (MCA) antibodies to type- and group-specific antigenic determinants of HSV-1 and HSV-2. Seven of them were studied at length by western blot and radioimmunoprecipitation methods. The cell lines 1H97 and 2H141 were shown to produce immunoglobulins of G2 beta and M class, respectively, and were directed against group-specific antigenic determinants of the major nucleocapsid protein p150. The cell lines 1H38 and 1H110 produced immunoglobulins of M and G2 beta, respectively, and were directed against type-specific antigens of HSV-1 glycoprotein gB. At the same time, the presence of group-specific antigenic determinants on glycoprotein gB molecule was indicated by MCA 1H188 belonging to immunoglobulins of G2 alpha class. Two cell lines, 2H208 and 1H225, produced immunoglobulins G2 alpha directed against type-specific antigenic determinants of HSV-2 glycoprotein gD and group-specific antigenic determinants of HSV-1 gD, respectively. The results of immunoelectron microscopy indicated that MCA 1H110 and 2H208 were directed against virus envelope proteins.
Asunto(s)
Anticuerpos Monoclonales/aislamiento & purificación , Simplexvirus/inmunología , Proteínas Virales/inmunología , Animales , Anticuerpos Monoclonales/análisis , Línea Celular , Núcleo Celular/inmunología , Reacciones Cruzadas , Citoplasma/inmunología , Epítopos/inmunología , Epítopos/aislamiento & purificación , Femenino , Hibridomas/inmunología , Inmunización , Immunoblotting , Ratones , Ratones Endogámicos BALB C , Microscopía Inmunoelectrónica , Simplexvirus/aislamiento & purificación , Proteínas Virales/aislamiento & purificaciónRESUMEN
Eight recombinant clones were obtained by insertion of BamHI fragments of herpes simplex type I viral DNA into a vector plasmid pUC19o. Of the obtained clones 5 were found to hybridize with herpes simplex type I and 2 viral DNA while 3 clones revealed a positive reaction with the Vero cells DNA. A constructed DNA-probe possessing the highest level of activity was selected for further studies. The probe is a BamHI fragment of herpes simplex type I viral DNA labelled with 32P dTTP. Probe sensitivity in blot hybridization is 10 pg for identification of type I viral DNA and 50 pg for type 2 viral DNA. The DNAs of cytomegalovirus and herpes zoster virus do not show positive signals with the probe. The increased sensitivity of the used dot hybridization as compared with biological or IEA antigen identification of the virus was confirmed with the clinical material from 59 patients with the different clinical manifestations of the herpes viral infection.