Journal of General Virology - Volume 2, Issue 3, 1968
Volume 2, Issue 3, 1968
- Articles
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The Origin of Hybrid Variants Derived from Subtype Strains of Foot-and-Mouth Disease Virus
More LessSummaryTwo strains of foot-and-mouth disease virus, distinguishable by four marker characteristics in addition to their antigenic subtype, could be separated by countercurrent distribution in an aqueous polymer phase system. This technique of separation was applied to the analysis of the progeny virus obtained from pig kidney cells simultaneously infected with these two strains.
Selection for recombinant virus yielded a number of isolates with the appropriate recombinant phenotype. These isolates subsequently segregated into a number of clones of various phenotypes including parental and recombinant types. Possible interpretations of this phenomenon are discussed. It is suggested that the segregating isolates may have originated from foci of infection inititated by viral aggregates.
The serological data suggest that the frequency of recombinants in the progeny may be greater than indicated by multiple marker exchanges.
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The Serological Relationship of the Soluble Antigens of Adenovirus Type 19 *
More LessSummaryThe relationship of group-specific (hexon) antigen, type-specific (fibre) antigen, soluble haemagglutinin (dodecon) and purified virus particles of adenovirus 19 was studied by cross complement-fixation, haemagglutination-inhibition and neutralization with antisera prepared against purified antigens. The fibre antigen reacted as an indirect haemagglutinin, producing haemagglutination in the presence of selected heterologous adenovirus immune sera. Dodecon and fibre antigen were closely related, while hexon antigen showed no relationship to fibre and possibly a weak relationship to dodecon antigen. Neutralizing antibodies were found in antisera to hexon and dodecon, but not in fibre antisera.
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Complete and Abortive Infection of Cell Cultures by Influenza A2 Virus
More LessSummaryThe reproduction of a mutant of A2/singapore/57, selected after repeated passages in monkey kidney cells, and the synthesis of its subviral components were followed in two systems: in monkey kidney cells, in which new infectious virus is formed, and in human diploid cells, in which the growth cycle of this virus is abortive. The formation of subviral antigens was investigated by both immunofluorescence and complement-fixation tests. No marked differences were found between the kinetics of synthesis of the V and S antigens and their total yields in both systems. Migration of the S antigen from the nuclei was observed in A2/singapore virus infected by human diploid cells.
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The Mechanism of Ultraviolet Induction in Lysogenic Staphylococcus aureus
More LessSummaryThis investigation has been focused on the significance of residual growth in the latent period after ultraviolet irradiation of a lysogenic strain of Staphylococcus aureus (111). It was shown that the organisms divide once or twice before the development of free phage and the number of infective centres are increased by about 100% in 30 min. after irradiation.
When celbenin (sodium 6-(2,6 dimethoxy benzamido) penicillinate) was added to the culture after irradiation, the number of infective centres decreased progressively, and no free phage developed. A delay of 10, 20 or 30 min. in introduction of the drug allowed an increasing fraction of the infective centres to develop and to yield free phage.
These observations lead to the suggestion that ultraviolet irradiation inhibits the synthesis of new repressor of vegetative phage development, but has no influence on repressor molecules present beforehand. Residual growth of the irradiated bacteria serves to dilute the intracellular repressor concentration below an effective level.
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Characterization of Herpes Simplex Virus Strains Differing in their Effects on Social Behaviour of Infected Cells
More LessSummaryEstablished (laboratory) strains and fresh isolates of herpes simplex virus from patients with skin and genital lesions were classified into four groups depending on their effects on the social interaction among infected hep-2 cells. The groups comprised strains causing (1) rounding of cells but no adhesion or fusion, (2) loose aggregation of rounded cells, (3) tight adhesion of rounded cells, and (4) fusion of cells into polykaryocytes. Protype strains from each group were found to differ with respect to immunologic specificity, buoyant density in CsCl solutions and stability at 40°.
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The Intracellular Site and Sequence of Sindbis Virus Replication
More LessSummaryThe replication of Sindbis virus was studied in primary chick embryo fibroblasts treated with actinomycin D. The cellular site of viral RNA and coat protein synthesis was found to be localized in the cytoplasmic reticulum. Two different species of RNA were identified in association with the reticulum, namely a double-stranded RNA with a sedimentation constant of 20 S and RNA with a sedimentation constant of 25 S. The single-stranded RNA molecules extracted from the 130 S virus particles had a sedimentation coefficient of 40 S. The time course of the association of viral RNA and coat proteins demonstrated that both components appear simultaneously in the 130 S viral ribonucleoprotein particles within 15 min.
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Immunofluorescent Studies on the Inhibition of Influenza A and B Viruses in Mammalian Cell Cultures by Amines and Ammonium Compounds *
More LessSummaryA quantitative immunofluorescent cell-counting technique was used to investigate the virustatic effect of aminoadamantane, ammonium acetate and a number of aliphatic amines on the development of influenza virus antigens in BHK-21 cell monolayers. Influenza virus strains A2/scotland/49/57, A/nws and B/england/939/59 were used at high multiplicities of infection in the tests. Quantification of the activity of the antiviral compounds was provided by the direct estimation of the proportion of infected cells in which the production of influenza virus fluorescent antigens was blocked. Comparable results were obtained for A2/scotland/49/57 virus using the immunofluorescent technique and the more conventional method of measuring antiviral activity by the reduction in the ability of the virus to multiply in tissue culture in the presence of the test compounds.
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Serological Relationships between the Neuraminidases of Influenza Viruses
More LessSummarySerological relationships between neuraminidases of representative strains of influenza virus isolated between 1930 and 1967 were studied, using rabbit antisera. The neuraminidases of type A strains formed three antigenic groups, sw, A0–A1 and A2, with antigenic drift within these groups. The neuraminidases of type B strains were unrelated to those of type A strains, and also showed antigenic drift. Four subgroups could be distinguished in haemagglutination-inhibition tests, sw, A0, A1 and A2. Antigenic drift occurred within each of these subgroups; antigenic changes in the neuraminidase and haemagglutinin occurred independently.
The neuraminidases of different strains as well as variants of a single strain varied greatly in their heat stability, which had no direct relation to the antigenic changes or the time of isolation of the strains. Both the antigenicity and the enzymic activity of the neuraminidases of certain strains (nsw, wse) were heat-labile and it was necessary to use freshly prepared virus suspensions for preparing antisera to their neuraminidases.
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Aggregated Forms of the Satellite of Tobacco Necrosis Virus
More LessSummaryThe several kinds of the two- and three-dimensional crystalline forms produced by the satellite virus of tobacco necrosis virus are described and discussed.
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The Largest Rabies-specific Antigen in Extracts of Infected Suckling Mouse Brains
More LessSummaryExtracts of rabies-infected suckling mouse brains purified by precipitation at pH 4.5, freed from smaller antigens by sedimentation at 161, 180g and digested with RNase, DNase and trypsin show in the ultracentrifuge a component of S 20 ≈ 16 to 18 which is lacking in extracts of normal suckling mouse brains similarly treated. The largest rabies soluble antigen (‘outer antigen’: Mead, 1962b) has a sedimentation constant S 20 ≈ 16 estimated by the ‘biological’ method of Polson & van Regenmortel (1961). The purified antigen appears to consist of rings or possibly single-turn helices about 100 Å in diameter containing about 0.57 µg. pentose (as ribose) per µg. total nitrogen. The antigen also appears to contain deoxypentose. It is resistant to pancreatic RNase, DNase, trypsin and chymotrypsin, has a density of about 1.34 g./cm3. in CsCl and an electrophoretic mobility about 7/8 that of rabbit serum albumin at pH 8.5.
Preparative density-gradient centrifugation in the analytical rotor of the Model E Spinco centrifuge is described. This allows the method to be applied to smaller particles than can be treated in the S.W. 39 rotor.
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Some Electron Microscopic Studies on the Satellite Tobacco Necrosis Virus and its IgG-antibody
More LessSummarySatellite Tobacco Necrosis Virus is the smallest virus known to date. It has a mean diameter of 180 Å with icosahedral symmetry which was determined from shadow-cast and negatively contrasted specimens. Immunological complexes of virus and anti-virus (IgG) were freed from non-specific IgG by particle sieve chromatography. The individual antibody molecules could be detected by electron microscopy in the small complexes formed with slight antigen excess. IgG molecules either bridged adjacent virus particles by their ends or were combined to a single virus particle by one of its ends, thus radiating out from the virus capsid. A few subunits could be discerned on the IgG molecules arranged in different structural configurations.
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The Effect of 5-fluoro-deoxyuridine on the Replication of Viral DNA and Synthesis of Polyoma Capsid Protein
More LessIt has been reported that synthesis of specific viral protein requires replication of viral DNA. Various investigators (Flanagan & Ginsberg, 1962; Kjellén, 1962; Gilead & Ginsberg, 1965) found that the formation of virus specific proteins in adenovirus-infected cells was directly dependent upon the synthesis of viral DNA. The analogue 5-fluoro-2-deoxyuridine (FUdR) completely inhibited synthesis of viral antigen in human foetal muscle cells infected with cytomegalovirus (Goodheart, Filbert & McAllister, 1963) and suppressed the production of specific virus antigen in SV40 infected monkey kidney cells (Melnick, Stinebaugh & Rapp, 1964; Gilden et al. 1965). In contrast, the work on pseudorabies virus (Reissig & Kaplan, 1962) suggested that specific viral protein formation can proceed in the absence of apparent viral DNA replication and synthesis of viral protein under conditions of FUdR inhibition has been reported for vaccinia virus (Shatkin, 1963; Salzman, Shatkin & Sebring, 1963; Loh & Payne, 1965) and polyoma virus (Sheinin, 1964).
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The Widespread Occurrence of Enteric Flagellar Phages
More LessThe first phage known to adsorb to bacterial flagella was isolated by Sertic & Boulgakov (1936a), who found it attacked motile but not non-motile strains of various enterobacteria. Since then, such phages have only been reported in 1941, when three isolates were obtained from the East River, New York, by Rakieten & Bornstein (1941). Flagellar phages nevertheless appear to be very common in sewage, for we have found them in 5 of 18 samples examined for I phages (Lawn et al. 1967). All the new phages are closely related to the original flagellar phage named x (Sertic & Boulgakov, 1936b), which we therefore propose to name x1, the new phages being x2 to 6 as the phages isolated by Rakieten & Bornstein (1941) are no longer available (M. L. Rakieten, personal communication).
The method of selection consisted of incubating 1 ml. of sewage, clarified by centrifugation for 15 min. at 700 g, with 9 ml.
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Influence of Animal Genotype and Age on the Amount of Circulating Interferon Induced by Newcastle Disease Virus
More LessFollowing the paper by Baron & Buckler (1963) describing the release of interferon (Isaacs & Lindenmann, 1957) into the blood of mice injected with viruses, extensive use has been made of this experimental procedure to study various aspects of interferon synthesis in vivo. The usual criterion for selecting animals for such experiments has been to use those within a narrow weight range in random-bred strains. During an investigation on the effect of carcinogens on interferon synthesis in mice (De Maeyer & De Maeyer-Guignard, 1967) we noticed that the amounts of circulating interferon in animals injected with Newcastle disease virus varied with animal age and strain. This observation led to more systematic experiments which are reported here.
Mice belonged to three strains. C3H/He and C57Bl mice were purchased from the Laboratory Animal Centre (M.R.C., Carshalton, Surrey, England). Only the first and second generations derived from the Carshalton breeder mice were studied.
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Haemadsorption by Vaccinia Virus and the Vh Factor in Chickens
More LessFowl red cells are agglutinated by vaccinia virus, but the mechanism is different from the haemagglutination by myxoviruses. Clark & Nagler (1943) made an extensive study of this phenomenon, and investigated different races of chickens at different ages. They pointed out that the red cells of newly hatched chickens were consistently negative, but that from older birds about 50% were negative in the vaccinia haemagglutination reaction, regardless of age and race. Burnet & Stone (1946) found that red cells agglutinated by vaccinia virus were also agglutinated by cardiolipin and other lipids. Suzuki et al. (1955) extended this work, and showed that while red cells of most White Leghorns were positive, the cells of Plymouth Rock chickens were negative in the vaccinia haemagglutination test.
Gilmour (1959, 1960, 1962,) in an extensive study of blood groups in fowls, considered the property of being agglutinated by vaccinia virus as a separate blood group Vh which was certainly not linked with three other blood groups (C, L and N) and probably not with a further two (A and B).
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The Use of Batch-type Zonal Ultracentrifuge Rotors for the Isolation and Purification of Viruses
More LessContinuous-flow zonal ultracentrifugation is now established as an invaluable technique for rapidly concentrating, and largely purifying, milligram amounts of infectious virus from very large volumes of tissue culture fluid (Cline, Nunley & Anderson, 1966; Toplin, 1967; Cline et al. 1967; Fox et al. 1967). The quantities of virus thus obtainable are potentially of use for chemical and physical studies of virus nucleic acid and protein or for production of vaccines. However, for many purposes, sufficient virus can be obtained from volumes of tissue culture fluid of the order of 1 l., whereas continuous-flow zonal rotors are most suited for handling volumes of 5 l. and upwards. Moreover, a continuous-flow rotor is a specialized piece of equipment and so is available in relatively few laboratories. Batch-type zonal rotors, on the other hand, are much more widely available since they are used for separating a variety of subcellular components (Anderson et al. 1967).
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Rescue of Rous Sarcoma Virus in Mixed Cultures of Virogenic Mammalian and Chicken Cells, Treated and Untreated with Sendai Virus and Detected by Focus Assay
More LessRous sarcoma virus could be detected in mammalian tumours induced by it if these tumours were transmitted as fresh tissue suspensions into chicks (Svoboda, 1960, 1961). Detailed studies of this question (Svoboda, 1962, Šimkovič, Valentová & Thurzo, 1962; Svoboda et al. 1963) have established that close contact between virogenic mammalian cells and chicken cells is necessary for the formation of Rous sarcoma virus, and fusion between these cells is regarded as a most likely mechanism enabling the transmission of Rous sarcoma genetic material from the mammalian cells to the chicken cells (Svoboda et al. 1963). The finding that Sendai virus produces fusion between homologous (Okada, 1962) and heterologous (Harris & Watkins, 1965) cells stimulated us and others to make experiments which showed that the procedure also significantly increased the production of Rous virus in mixed cultures (Svoboda, Machala & Hložánek, 1967; Vigier, 1967; Yamaguchi, Takeuchi & Yamamoto, 1967).
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Observations on Complete and Empty Capsids of Foot-and-Mouth Disease Virus
More LessRecent reports have described the similarities and differences in the composition and occurrence of infectious virions and empty capsids (or top components) of several small RNA viruses (Maizel, Philips & Summers, 1967; Longley & Leberman, 1966; Fabiyi, Engler & Martin, 1964; Halperen, Eggers & Tamm, 1964). A previous report (Breese & Graves, 1966) on observations of crystalline arrays of foot-and-mouth disease virus type A, strain 119, has been followed by extensive experimentation with three field strains from Argentina (A1, O2 and C3, CANEFA), which have had fewer tissue culture passages. An effort was made to find a combination of multiplicity of infection, time of harvest, and cell culture which would regularly result in the observation of crystalline arrays of virus particles. During the course of these studies, the A1 strain was found to form arrays of both complete and recently described empty particles.
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Analysis of Nearest Neighbour Base Frequencies in the RNA of a Mammalian Virus: Encephalomyocarditis Virus
More LessThe technique of frequency analysis of nearest neighbour base sequences in DNA (Josse, Kaiser & Kornberg, 1961) allows the characterization and description of a DNA in terms of the average frequency of occurrence of its sixteen possible doublet sequences. This technique has recently been used to study the patterns of doublet frequencies before and after normalization in the DNA of nine mammalian viruses (Subak-Sharpe, et al. 1966; Morrison et al. 1967; Subak-Sharpe, 1967). It was found that the doublet pattern in the DNA of the four small oncogenic viruses studied resembled the pattern in the host DNA to a remarkable extent, whereas the five large viruses investigated showed virtually no similarity to the host pattern.
The authors reasoned a priori that a cell's translation apparatus, being the result of natural selection, would have a population of transfer RNAs adapted optimally to translate the polypeptide-specifying sequences of the cell DNA.
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