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Biomedical subjects

K R Dumbell

Publications and source records attributed to K R Dumbell.

18 recordsLinked to original sources

Two new genome types of adenovirus 7c.

Adenovirus type 7 is the type most frequently associated with serious disease. Eighteen different genome types of adenovirus type 7 had been reported up to October 1986. The genome type Ad7c, based on the restriction enzyme profiles of SmaI and BamHI, has been reported from Europe prior to 1969 and more recently from South Africa. Here, we report two new genome types of adenovirus 7 c that have not previously been identified and that have been isolated in South Africa between 1975 and 1986 from children with postmeasles pneumonia. The two new genome types differ from the prototype Ad7c virus in having two (Ad7c1) or one (Ad7c2) extra cleavage sites for the restriction endonuclease EcoRI. These sites have been located at 3.68kb and 5.32kb from the left terminus of the genome map published for the prototype Ad7c strain. A strain resembling the prototype Ad7c was also isolated in 1986 from a case of post measles pneumonia.

Adenoviridae Infections

Characterisation of poxviruses from sporadic human infections.

An orthopoxvirus was isolated from the vesicular rash of a man in Natal who died in coma and who had not been vaccinated. Analysis of the viral DNA showed that it was a vaccinia virus, more closely related to the virus of South African smallpox vaccine than to other vaccinia viruses. DNA analysis also showed that an orthopoxvirus isolated from a sporadic case of severe pustular rash in Nigeria was a vaccinia virus closely related to the smallpox vaccine virus used there. Minor biological differences between the viruses isolated and the corresponding vaccine strains suggested that some natural transmission of the virus had occurred, but the results of DNA analysis implied that they originated from the use of smallpox vaccine. No similar cases have been detected since smallpox vaccination was discontinued.

Adult

The virology of variola minor. Correlation of laboratory tests with the geographic distribution and human virulence of variola isolates.

Several groups of variola isolates were compared in DNA structure, and by four independent biologic markers. Isolates of variola minor from Europe and South America (alastrim virus) could be distinguished from African isolates of variola minor by DNA structure and by two of the four biologic markers. Taken as a group, the properties of African isolates, in general, differed from those of variola major, but this difference was confined to properties which depended (in the laboratory) on the recent history of the virus concerned. The suggestion made previously that there was an "intermediate" or "African" variety of variola virus is discounted. Laboratory tests did not distinguish any individual African isolate from variola major virus. It is concluded that a virus which may be called "alastrim" represents a "fixed" variant of variola virus, whose distribution is consistent with the dramatic spread of variola minor through the Americas and Europe in the early part of this century, and that variola minor in Africa in recent years was due to variola virus which was not alastrim and which laboratory evidence fails to identify as an entity distinguishable from variola major virus.

Africa, Western

The genome structure of cowpox virus white pock variants.

A previous report described restriction endonuclease analysis of white pock variants of red cowpox virus and their characterization as deletion mutants lacking certain sequences including the repetition from one specific terminus of the wild-type genome. Further analysis has confirmed the terminal deletion but demonstrated that this is compensated at the site of deletion by the presence of an inverted duplication of a variable amount of sequence from the opposite terminus, with the effect of restoring a terminal repetition and the covalent, terminal crosslink. Nine of 11 white pock variants showed a similar deletion of about 21 Mdal mapping contiguously from the right-hand terminus and extending into a 2.4 Mdal restriction fragment. Two white variants showed larger deletions of about 24 and 27 Mdal respectively. These deletions were compensated by a copy of sequences from the opposite terminus which ranged in size from 3 to 27 Mdal. No terminal deletions smaller than 21 Mdal were observed in cowpox white variants or in clones retaining the red phenotype. In contrast with other orthopoxviruses, no deletions involving the left-hand terminus were found. Some independent white isolates had similar sizes of sequence copied from the opposite terminus, but some sibling clones from a single, pock-purified white isolate with the same size of deletion had different sizes of duplicated sequence. Other siblings isolated from an independent, three times pock-purified white clone, itself derived from a single parental red pock, differed from each other in the size of both the deletion and the duplicated sequence. These observations suggest preference for deletion in a particular region, conjunction of the genome termini during DNA replication and a requirement for the preservation of symmetrical termini in orthopoxvirus genome function.

Base Sequence

A poxvirus antigen associated with pathogenicity for rabbits.

White pock variants of cowpox virus give papular lesions on intradermal inoculation of rabbits, without the necrosis and haemorrhage that are produced by wild type cowpox viruses. Rondle & Dumbell (1962) have shown that white pock variants of cowpox virus fail to produce a specific, precipitating antigen which they called 'd' substance. In this paper it is shown that 'd' is demonstrable in soluble antigen preparations of rabbitpox virus and of neurovirulent strains of vaccinia virus but not in soluble antigens of variola viruses. Two series of recombinant viruses prepared by Dumbell & Bedson (1964) from variola and cowpox and from variola and rabbitpox viruses were tested for the production of 'd' substance. These results were compared with the previously recorded effects of these recombinants when inoculated intradermally in rabbits. It is concluded that functional genes determining the production of 'd' and of rabbit skin pathogenicity are closely linked on the pox virus genome, but that there is insufficient evidence to say that the two functions are interdependent.

Animals

The pathogenicity of variola virus. A comparison of the growth of standard strains of variola major and variola minor viruses in cell cultures from human embryos.

The international reference strains of variola major (Harvey) and of variola minor (Butler) were grown in cultures of skin and muscle cells from human embryos. The development of infective virus, complement-fixing antigen, haemagglutinin and cytological changes were followed at four temperatures between 35 and 40 degrees C. No significant difference was found in the amount of virus produced by Harvey or Butler viruses at any of the experimental temperatures, but Harvey attained the plateau titre at 16 h, some 4 h ahead of Butler in the cultures incubated at 38 degrees C. Harvey also produced a higher and more prolonged yield of virus in the extracellular medium of cultures, inoculated at low multiplicity and incubated at 37 degrees C. At 38 degrees C small inocula of Harvey produced foci which developed and spread till the whole culture was necrotic; Butler foci did not spread and remained relatively undeveloped at this temperature.Staining with acridine orange showed the development of cytoplasmic inclusions at all temperatures up to 39.5 degrees C, at which temperature most inclusions remained round and discrete and susceptible to digestion with DNase. The yield of virus at all temperatures correlated well with the number of cells in which the DNA cytoplasmic inclusions became irregular in outline, diffuse and insusceptible to digestion with DNase. It was concluded that elevated temperatures, up to 39.5 degrees C affected principally a maturation phase in the development of the virus.Equal amounts of complement-fixing antigen were produced at all temperatures and by either virus, but the late product, haemagglutinin was depressed at elevated temperatures much more in cultures infected with Butler than in cultures infected with Harvey. This was clearly shown by haemadsorption; in cultures, infected at high multiplicity and incubated at 39.5 degrees C Harvey gave semi-confluent haemadsorption, while only an occasional haemadsorbing cell could be found in the cultures infected with Butler virus.It is concluded that there were two ways in which temperature affected the growth of variola viruses in cultures of human embryo skin and muscle cells. The total yield of virus was reduced by inhibition of a stage in the maturation of the virus with a cut off point between 39.5 and 40 degrees C at which no cells produced infective virus, and with little observable differences between Harvey and Butler viruses. Changes at the surface of virus-infected cell, involving virus release and haemagglutinin, were affected independently of virus maturation; in these changes Butler was much more sensitive than Harvey to elevated temperatures. The relevance of these observations to the development of variola major and variola minor in man is discussed.

Cells, Cultured

Laboratory investigation of two "whitepox" viruses and comparison with two variola strains from southern India.

Two variola-like viruses were isolated in Bilthoven in 1964 from monkey kidney tissue cultures. These viruses, coded 64/7255 and 64/7275, have been considered as two of the six "whitepox" viruses isolated from animal tissues, all of which are indistinguishable from variola virus by laboratory tests.Two specimens from suspect smallpox cases in India were examined in the Bilthoven laboratory at about the same time as the "whitepox" viruses and two strains of variola virus were isolated from them. A detailed comparison of certain biological markers of these four viruses, and of their DNAs, shows that the two "whitepox" viruses could not be distinguished from each other or from one of the two variola isolates. In view of this, and since there was a possibility of cross-contamination at the time of isolation, it is concluded that 64/7255 and 64/7275 must be regarded as genuine variola viruses and be deleted from the list of variola-like viruses isolated from animal tissues.

DNA Restriction Enzymes

Comparison of white pock (h) mutants of monkeypox virus with parental monkeypox and with variola-like viruses isolated from animals.

Monkeypox mutants arising spontaneously or after serial, high multiplicity passage were characterized phenotypically and by restriction endonuclease mapping. Some resemble "whitepox" and variola viruses in several of the markers tested but all are distinguishable phenotypically from these. None resembles "whitepox" viruses in genome structure although near-terminal deletions or symmetrical, terminal rearrangements, relative to parental monkeypox, occurred. "Whitepox" viruses isolated from animals closely resemble variola in both phenotype and genome structure.

Base Sequence

Studies on Cotia virus--an unclassified poxvirus.

This paper is a report of studies on Cotia virus; this had been first isolated in 1965 in Brazil and was subsequently shown to be a poxvirus. Cotia virus grew in a wide range of cell cultures and on the chick chorioallantois (CAM), Its growth characteristics are similar to those of other poxviruses. Microscopy showed virus factories or type B inclusions appearing before infectious progeny virus could be demonstrated. Type A inclusions appeared later, after development of progeny virus; these were shown by electron microscopy to differ from the type A inclusions of cowpox and other poxviruses and they have been termed Cotia bodies. Immunofluorescent staining also showed ring structures which appeared before the development of Cotia bodies. The growth of Cotia virus in human embryo lung (HEL) cells was sensitive to inhibitors of DNA and protein synthesis but was resistant to a concentration of rifampicin which inhibited vaccinia virus. Sharing of antigens between the Cotia virus and vaccinia virus was shown by gel precipitation tests and immunofluorescent staining. There was no cross neutralization between Cotia virus and vaccinia virus nor did anti-Cotia sera neutralize representatives of other poxvirus groups.

Brazil

Concurrent smallpox and chickenpox.

Three patients showing smallpox- and chickenpox-like lesions simultaneously were investigated virologically. Both infections were confirmed in the laboratory and, in one case, by electron microscopy.

Adolescent

An unusual poxvirus from Nigeria.

A poxvirus of the variola-vaccinia subgroup was isolated from the lesions of a female African refugee suffering from a smallpox-like illness. The virus is interesting because it is neither variola nor vaccinia but combines some properties of each. These properties are described and the possible origin of the virus is discussed.

Adult