Asunto(s)
Antígenos/aislamiento & purificación , Proteínas del Sistema Complemento/efectos de la radiación , Virus de la Encefalitis/inmunología , Efectos de la Radiación , Animales , Reacciones Antígeno-Anticuerpo , Antígenos/efectos de la radiación , Embrión de Pollo , Cromatografía DEAE-Celulosa , Cobalto , Pruebas de Fijación del Complemento , Proteínas del Sistema Complemento/aislamiento & purificación , Técnicas de Cultivo , Estabilidad de Medicamentos , Virus de la Encefalitis Equina Venezolana/inmunología , Virus de la Encefalitis Equina Venezolana/efectos de la radiación , Virus de la Encefalitis/efectos de la radiación , Caballos , Dosificación Letal Mediana , Métodos , Ratones , Conejos , Factores de TiempoRESUMEN
Venezuelan equine encephalitis (VEE) virus was purified and concentrated by chromatography of tissue culture supernatant fluids on diethylaminoethyl-cellulose columns. Stepwise gradient elution studies indicated a broad elution pattern for the virus, with recovery occurring from 0.05 to 0.7 m NaCl. Optical density, infectivity, hemagglutination (HA), and complement fixation (CF) assays indicated that complete recovery of input virus in highly purified form was possible. Single-step elution with 0.7 m tris(hydroxymethyl)aminomethane-succinate-salt buffer resulted in a virus volume decrease of 85% with a concomitant increase in infectivity and antigenicity. Recoveries consistently equaled or exceeded 100% of the input preparations. Additional purification of column-recovered virus was obtained by sedimentation of pooled virus eluates on 50% sucrose cushions. Exposure of borate saline and 0.5% histidine suspensions of purified VEE virus preparations to 6 x 10(6) r of gamma radiation resulted in a loss of infectivity for tissue culture and a loss of lethality for weanling and suckling mice. Inactivation was an exponential function of the dosage. In contrast to infectivity, antigencity (HA and CF) of both saline and histidine preparations was retained after irradiation with doses as high as 6 x 10(6) r. Purified and irradiated VEE virus preparations have been successfully used for routine serological tests and are being evaluated as vaccines.
Asunto(s)
Virus de la Encefalitis/aislamiento & purificación , Virus de la Encefalitis/efectos de la radiación , Histidina/farmacología , Pruebas de Aglutinación , Animales , Línea Celular , Centrifugación , Embrión de Pollo , Cromatografía DEAE-Celulosa , Pruebas de Fijación del Complemento , Técnicas de Cultivo , Virus de la Encefalitis/efectos de los fármacos , Virus de la Encefalitis/patogenicidad , Encefalomielitis Equina/microbiología , Cobayas , Técnica de Placa Hemolítica , Ratones , Efectos de la RadiaciónRESUMEN
Hemagglutinins of Venezuelan equine encephalomyelitis virus were produced in tissue cultures, and the infective virus was rendered nonviable by exposure to gamma radiation.