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Extended interhuman transmission of monkeypox in a hospital community in the Republic of the Congo, 2003.
This report describes the first reported outbreak of human monkeypox in the Republic of Congo. Eleven confirmed and probable monkeypox cases were observed during this outbreak, all were less than 18 years old, and most resided on the grounds of the Government Hospital in Impfondo. Molecular, virologic, and serologic, and diagnostic assays were used to detect evidence of monkeypox (or orthopox) virus infection in individuals with striking dermatologic and other clinical manifestations. The majority of cases in this outbreak experienced significant, symptomatic illnesses; there was one death, possibly involving secondary complications, and one instance of profound sequelae. Up to six sequential transmissions of monkeypox virus from person to person are hypothesized to have occurred, making this the longest uninterrupted chain of human monkeypox fully documented to date. The pattern of sustained human-to-human transmission observed during this outbreak may influence our current perception of the capacity for this zoonotic virus to adapt to humans.
[Monkeys as a source of human viral diseases].
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Adjuvant-enhanced antibody responses to recombinant proteins correlates with protection of mice and monkeys to orthopoxvirus challenges.
Recombinant proteins are being evaluated as smallpox and monkeypox vaccines because of their perceived safety compared to live vaccinia virus. Previously, we demonstrated that three or more injections of a Ribi-type adjuvant with a combination of three proteins from the outer membranes of intracellular (L1 protein) and extracellular (A33 and B5 proteins) forms of vaccinia virus protected mice against a lethal intranasal challenge with vaccinia virus. Here, we compared several adjuvants and found that QS-21 and to a lesser extent alum+CpG oligodeoxynucleotides accelerated and enhanced neutralizing antibody responses to a mixture of L1 and A33 proteins, provided the highest ratio of IgG2a to IgG1 isotype response, and protected mice against disease and death after only two immunizations 3 weeks apart. In addition, monkeys immunized with recombinant vaccinia virus proteins and QS-21 developed neutralizing antibody to monkeypox virus and had reduced virus load, skin lesions, and morbidity compared to the non-immunized group following monkeypox virus challenge.
Isolation of poxvirus from an African Rodent.
A poxvirus was isolated from a wild gerbil (Tatera kempii) caught in northern Dahomey, Africa at the time of an epidemic of human smallpox. Electron microscopic appearance and serologic reactions placed it in the vaccinia subgroup of poxviruses. The isolate differed from ectromelia, rabbitpox, vaccinia, monkeypox, and cowpox viruses in pock morphology on chorioallantoic membrane, ceiling temperature, relative innocuity for mice, and cytopathic effect in tissue culture. Like variola minor virus, it had a ceiling temperature of 38 C, produced small hypertrophic foci in tissue culture, and failed to grow in rabbit skin. Inoculated into a rhesus monkey, it caused fever but no skin eruption and produced seroconversion and protection from subsequent challenge with monkeypox virus. The growing list of animal viruses that differ only slightly from smallpox virus suggests the hypothesis that long-term survival of variola virus may be based on inapparent infection in animals as well as virulent spread among humans.
The agents of biological warfare.
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Bioluminescence Imaging to Study Recombinant Orthopoxvirus Infection in Animal Models.
Bioluminescent images of viral replication in live animals (in vivo) reveal disease dynamics and effects of medical countermeasures over time. After selecting an appropriate orthopoxvirus animal model for the study, a recombinant virus with the firefly luciferase gene inserted in the genome is used to infect the animals. On the day of bioluminescent imaging, the substrate, D-luciferin, is prepared; animals are sedated and injected with the substrate and IVIS imager is utilized; various bioluminescent images are acquired; then animals recover and are able to continue in the study. Ex vivo imaging can also be completed after animals are euthanized at experimental endpoint. This approach allows real-time imaging of viral kinetics within an animal, and analysis of images can provide an additional quantitative measure throughout the study. Bioluminescent imaging not only provides scientific benefits but also benefits to animal welfare. For these reasons, bioluminescent imaging should be considered for any in vivo orthopoxvirus study.
Experimental infection of squirrels Sciurus vulgaris by monkey pox virus.
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Is an old virus up to new tricks?
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Should the smallpox virus be allowed to survive?
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[Recent smallbox epidemics in Ethiopia and Somalia, human monkey pox virus infections and impossibilities of world eradication of the smallpox].
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A method for serological differentation of closely related poxviruses.
A two-step gel precipitation reaction with antiserum to vaccinia virus, based on sequential addition of two test viruses was elaborated for differentiation of closely related orthopoxviruses. The appearance of an additional precipitation line indicated differences in the antigenic structure of the viruses compared. Differences in the antigenic structure of vaccinia, variola and monkey pox viruses as well as the antigenic identity with cow pox virus of poxviruses isolated from carnivora, elephants and okapi were established.
A Quantitative Real-Time PCR Assay for Measuring Poxvirus Replication and Cell Binding.
Quantitative real-time PCR (qPCR) is a fast and reliable method to quantify viral genomes as a surrogate to titering on monolayers of cells for measuring virus replication. Whether it be for determining the number of virions released, the total number of genomes produced during infection, or the number of virions bound to a cell, qPCR assays can be adapted to quickly enumerate total viral genomes in a broad range of experiments comparing virus replication under different conditions. In addition, qPCR offers several advantages compared to plaque assays including time, linearity over 9 logs, and scalability from tens-to-hundreds of samples, depending on the qPCR machine. Here we describe a qPCR assay for quantifying vaccinia virus' dsDNA genome that can be used to determine the total number of virions produced. Furthermore, we describe a straightforward protocol for a cell-binding assay that is sensitive enough to use with small concentrations of inoculating virions. This protocol is suitable for measuring the cell-binding ability of mutations that affect virus production and infectivity.
Evolutionary history of orthopoxvirus proteins similar to human complement regulators.
Orthopoxviruses include many important pathogens such as variola major virus, camelpox, buffalopox, monkeypox, cowpox, and variola minor viruses. This group of viruses also includes vaccinia virus, which is extensively used in human vaccine development. Genomes of orthopoxviruses encode proteins with sequences similar to human regulators of complement activation (RCA) that contain tandem short consensus repeats (SCRs). We employed phylogenetic tree analysis to evaluate the structural relationships among SCRs of orthopoxvirus RCA-like proteins and those of human complement regulators. The human complement RCA proteins analyzed were factor H (FH), C4 binding protein alpha chain, membrane cofactor protein (MCP), decay accelerating factor (DAF), and complement receptors type 1 (CR1) and 2 (CR2). Sequences of key poxvirus regulators of complement activation, vaccinia virus complement control protein (VCP), smallpox inhibitor of complement enzymes (SPICE), and cowpox inflammation modulatory protein (IMP) were similar to SCRs 1 through 5 of C4 binding protein, alpha chain, and they were also clustered with other homologous repeats of MCP, DAF, CR1, CR2, and FH. Phylogenetic clustering of RCA sequences suggested that poxvirus complement regulators VCP, SPICE, and IMP arose from a single ancestral sequence that shared similarity with all human regulators of complement activation. Any changes in poxvirus complement regulators leading to the enhancement of their ability to regulate complement activation likely resulted from new mutations in the viral lineages.
[Monkey pox virus diseases in humans].
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Public health. Dead virus walking.
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