<i>SECTION OF BIOLOGY</i>: I. A NEW AND RAPID METHOD FOR ISOLATING VIRUSES BY SELECTIVE FLUOROCARBON DEPROTEINIZATION*

Gessler, Albert E.; Bender, Charles E.; Parkinson, Martin C.

doi:10.1111/j.2164-0947.1956.tb00497.x

Cited by 153 publications

(27 citation statements)

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“…Stocks of avirutent Hampstead egg strain ectromelia virus (12) were obtained from infected chorioallantoic membranes of 14-day old chick embryos. Genetron extraction of crude ectromelia virus stocks was sometimes employed to obtain cleaner virus suspensions (15). Semliki Forest virus stock was an homogenate of infected mouse brain in saline.…”

Section: Methodsmentioning

confidence: 99%

Mechanisms of Recovery From a Generalized Viral Infection: Mousepox

Blanden

1970

The Journal of Experimental Medicine

159

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The response of a host animal to primary viral infection is complex, and the factors essential for recovery from the infection are difficult to define. Immune responses, both cell-mediated and hulnoral, interferon production, pyrexia, and local changes in acidity and oxygen tension in infected tissues have all been ascribed possible roles in recovery (1). The importance of any single mechanism probably varies in diseases of different pathogenesis or at particular times during the course of infection, and there will sometimes be interaction and synergism between individual factors.Since neutralizing antibodies and interferon have well documented modes of antiviral activity, they could logically be assigned rather more importance than cell-mediated immunity, which has no clearly demonstrated antiviral mechanism. However, an increasing body of evidence derived from the study of human infants with primary immunological deficiency diseases points to a very important, perhaps even essential role for a thymus-dependent, cellmediated immune response in recovery from vaccinia virus infection (2). Experimental evidence concerning this area of host-virus relations is meager, but recently Hirsch et al. (3) showed that heterologous anti-thymocyte serum (ATS) 1 caused significantly increased mortality and morbidity from intravenous vaccinia infection in mice. They postulated that this was due to suppression of cell-mediated immunity; however no direct evidence was produced to show that the cell-mediated immune response to vaccinia virus was suppressed by ATS. This was assumed on the basis of an immunosuppressive potency test usiag lymphocytic choriomeningitis virus (4).Previously, Allison and Friedman (5, 6) had produced sound evidence that the regression of primary tumours caused by fibroma virus in rabbits was dependent upon cell-mediated immunity. However, this and other work coni Abbreviations use~ in this paper: AMS, rabbit anti-mouse macrophage serum; ATS, rabbit anti-mouse thymocyte serum; MEM, minimal essential medium; NRS, normal rabbit serum; PBS, phosphate-buffered saline; PFU, virus plaque-forming units; RBC, red blood cells WBC, white blood cells.

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Section: Methodsmentioning

confidence: 99%

Mechanisms of Recovery From a Generalized Viral Infection: Mousepox

Blanden

1970

The Journal of Experimental Medicine

159

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“…Arcton 113 (trifluorotrichloroethane, I.C.I. ; Gessler, Bender & Parkinson, 1956) by mechanical disruption in the chilled, stainless steel chamber of a Servall 'Omnimix'. The extract was centrifuged at 2,000 rev./min.…”

Section: Virusmentioning

confidence: 99%

The Influence of Some Metal Ions and pH on the Inactivation of Vaccinia Virus by Heat

Kaplan

1963

Journal of General Microbiology

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SUMMARYThe addition of metal ions to the suspending medium of vaccinia virus preparations made them more resistant to heat. Monovalent metals were more effective than divalent, but a mixture of the two kinds was the most effective. Heavy metals rapidly destroyed virus infectivity. The protective effect is probably due to the formation of metal-protein complexes with increased resistance to heat denaturation.

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“…Bender & Parkinson, 1956) was used in some of the extraction and purification procedures; it was obtained from Imperial Chemical Industries Ltd. as 'Arcton 63'.…”

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The Infectivity of Purified and Partially Purified Preparations of Vaccinia and Cowpox Viruses

Kaplan

Valentine

1959

Journal of General Microbiology

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SUMMARY: Twenty-three preparations of vaccinia virus and 5 of cowpox virus were examined by electron microscopy of spray droplets and by infectivity titrations at various stages of purification. Extraction of virus-containing tissues with a fluorocarbon compound was a useful early step in purification, but preparations of the highest purity were obtained by flocculation of the virus with M-NaCl. The average ratio of total particle count to infectivity for the vaccinia virus preparations was 12. The lowest observed ratio based on reliable particle counts was 3.4.During inactivation of vaccinia virus, a very small, but constant, fraction of infectivity was observed t o survive extremely high doses of gamma-radiation .In order to assess the likelihood of multiple reactivation being responsible for this persistence, it was necessary to know the ratio of infective units to total particle count in the type of virus preparation being irradiated. The two groups of published figures for vaccinia virus are not in close agreement, so several preparations of vaccinia virus, and a few of cowpox virus, were counted by electron microscopy and titrated for infectivity. The influence of various methods of purification on the preparations was also studied. METHODS Viruses.The vaccinia virus strains were all sublines of the Lister Institute vaccine strain. The rabbit dermal virus (RV) was a homogeneous line (Amies, 1938) in its 80th skin passage. Chick embryo adapted virus (LC) had been passed from 13 to 16 times in the chick embryo chorioallantois. Tissue culture virus (T) was taken from the first five batches of virus grown for smallpox vaccine preparation. This virus was a first passage of bacteriologically sterile RV in trypsinized chick embryo cells. The cozupoe virus was a strain lyophilized in 1939 and recently cultivated. Its history is unknown, but it produced typical haemorrhagic lesions in the chick embryo chorioallantois after 3 days of incubation.Diluent. All extractions and dilutions were made in McIlvaine's phosphate + citrate buffer (pH 7.2, 0.004 M-phosphate). Treatments with trypsin were done with Armour's crystalline, salt-free product.Extractions were made by mechanical disruption of the virus-infected tissues in a Servall 'Omnimix' homogenizer a t 14,000 rev./min. for 5 min., with an ice-bath round the stainless steel vessel. Trifluorotrichloroethane (Gessler,

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SECTION OF BIOLOGY: I. A NEW AND RAPID METHOD FOR ISOLATING VIRUSES BY SELECTIVE FLUOROCARBON DEPROTEINIZATION*

Cited by 153 publications

References 0 publications

Mechanisms of Recovery From a Generalized Viral Infection: Mousepox

Mechanisms of Recovery From a Generalized Viral Infection: Mousepox

The Influence of Some Metal Ions and pH on the Inactivation of Vaccinia Virus by Heat

The Infectivity of Purified and Partially Purified Preparations of Vaccinia and Cowpox Viruses

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