The genus Capripoxvirus within the family Poxviridae comprises three closely related viruses, namely goat pox, sheep pox and lumpy skin disease viruses. This nomenclature is based on the animal species from which the virus was first isolated, respectively, goat, sheep and cattle. Since capripoxviruses are serologically identical, their specific identification relies exclusively on the use of molecular tools. We describe here the suitability of the G-protein-coupled chemokine receptor (GPCR) gene for use in host-range grouping of capripoxviruses. The analysis of 58 capripoxviruses showed three tight genetic clusters consisting of goat pox, sheep pox and lumpy skin disease viruses. However, a few discrepancies exist with the classical virus-host origin nomenclature: a virus isolated from sheep is grouped in the goat poxvirus clade and vice versa. Intra-group diversity was further observed for the goat pox and lumpy skin disease virus isolates. Despite the presence of nine vaccine strains, no genetic determinants of virulence were identified on the GPCR gene. For sheep poxviruses, the addition or deletion of 21 nucleic acids (7 aa) was consistently observed in the 59 terminal part of the gene. Specific signatures for each cluster were also identified. Prediction of the capripoxvirus GPCR topology, and its comparison with other known mammalian GPCRs and viral homologues, revealed not only a classical GPCR profile in the last three-quarters of the protein but also unique features such as a longer N-terminal end with a proximal hydrophobic a-helix and a shorter serine-rich C-tail.3These authors contributed equally to this work.Two supplementary tables are available with the online version of this paper.Journal of General Virology (2009), 90, 1967-1977 DOI 10.1099 Davies, 1981Davies, , 1982Diallo & Viljoen, 2007). CaPV infections lead to similar clinical signs in sheep and goats, mainly characterized by fever, excess salivation, conjunctivitis and rhinitis with ocular and nasal discharges. These symptoms are followed by eruption of pox lesions throughout the skin. LSD is a subacute to acute cattle disease with appearance of skin nodules. During epizootics of CaPVs in domestic animals, disease in wild ungulates has never been reported (Davies, 1991), although serology and isolated cases suggests that wildlife species have a possible role in virus maintenance.CaPVs are generally considered to be host-specific, leading to outbreaks in one preferred host. This is partially true since some SPPV and GTPV isolates are capable of causing severe diseases in both sheep and goats (Davies, 1976;Kitching et al., 1989). An in-depth epidemiological investigation to specifically identify the viruses involved in acute small ruminant pox diseases cannot be achieved by serological testing alone due to the very close antigenic relationship between CaPVs, with the existence of only one serotype (Kitching et al., , 1989. The common immunogenic properties of these viruses have been used for the preparation of live attenuated vaccines that ...

To date, most HCA (High Content Analysis) studies are carried out with adherent cell lines grown on a homogenous substrate in tissue-culture treated micro-plates. Under these conditions, cells spread and divide in all directions resulting in an inherent variability in cell shape, morphology and behavior. The high cell-to-cell variance of the overall population impedes the success of HCA, especially for drug development. The ability of micropatterns to normalize the shape and internal polarity of every individual cell provides a tremendous opportunity for solving this critical bottleneck 1-2 .To facilitate access and use of the micropatterning technology, CYTOO has developed a range of ready to use micropatterns, available in coverslip and microwell formats. In this video article, we provide detailed protocols of all the procedures from cell seeding on CYTOOchip micropatterns, drug treatment, fixation and staining to automated acquisition, automated image processing and final data analysis. With this example, we illustrate how micropatterns can facilitate cell-based assays. Alterations of the cell cytoskeleton are difficult to quantify in cells cultured on homogenous substrates, but culturing cells on micropatterns results in a reproducible organization of the actin meshwork due to systematic positioning of the cell adhesion contacts in every cell. Such normalization of the intracellular architecture allows quantification of even small effects on the actin cytoskeleton as demonstrated in these set of protocols using blebbistatin, an inhibitor of the actin-myosin interaction.

Abstract:Because of their close relationship, specific identification of the CaPVs genus inside the Poxviridae family relies mainly on molecular tools rather than on classical serology. We describe the suitability of the G protein-coupled chemokine receptor (GPCR), for host range phylogenetic grouping. The analysis of 26 CaPVs shows 3 tight genetic clusters consisting of goatpox virus (GPV), lumpy skin disease virus (LSDV), and sheeppox virus (SPV).

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