Carnivore Protoparvovirus 1 at the Wild-Domestic Carnivore Interface in Northwestern Mexico.

Eighty-three wild and domestic carnivores of nine species from Janos Biosphere Reserve (JBR), Mexico, were tested by serologic and molecular assays to determine exposure and infection rates of carnivore protoparvovirus 1. Overall, 50.8% (33/65) of the wild carnivores and 100% (18/18) of the domestic dogs tested were seropositive for Canine protoparvovirus 1 (CPV), while 23% (15/65) of the wild carnivores and 22.2% (4/18) of the domestic dogs were PCR positive for CPV. Phylogenetic analysis confirmed circulation of CVP-2 with residues 426 Asn (CPV2a = 1/19) and 426 Glu (CPV-2c = 18/19) among carnivores in JBR. The prevalence of both PCR positivity and antibodies to CPV varied significantly among wild host species. Of the six identified haplotypes, three were unique to kit foxes (Vulpes macrotis) (the species with higher haplotype richness) and two to striped skunks (Mephitis mephitis). The remaining haplotype was shared among all carnivore species including dogs suggesting non-host specificity and bidirectional and continuous viral transmission cycle in the JBR. The phylogenetic similarity of CPV strains from dogs and wild carnivores and the higher prevalence of CPV in wild carnivores captured near towns relative to those captured far from towns suggest that dogs might be an important source of CPV infection for wild carnivores in the JBR. We provide evidence that cross-species transmission occurs at the domestic-wildlife interface in JBR.

Interleukin-8 mRNA synthesis and protein secretion are continuously up-regulated by respiratory syncytial virus persistently infected cells.

The aim of this study was to investigate whether respiratory syncytial virus persistence regulates interleukin 8 (IL-8) mRNA synthesis and protein secretion in a human lung epithelial cell line (A549). Therefore, we established RSV persistence in these cells (A549per) and determined the levels of interleukin-8 mRNA by RT-PCR and of protein through ELISA. Interleukin-8 mRNA synthesis and protein secretion were continuously up-regulated in A549per cells during passages and in A549 cells that had been incubated with supernatants (cA549per) obtained from A549per passages. These results suggested that the enhancement of interleukin-8 was stimulated either by the presence of the RSV genome in the cell or by soluble mediator(s) induced by RSV, which, in turn, increased interleukin-8 mRNA synthesis and protein secretion. Soluble RSV F and G proteins were identified as mediators. Moreover, interleukin-8 enhancement was observed after 1-min incubation with the soluble mediators, thus suggesting that interleukin-8 up-regulation was triggered by receptor-ligand interaction.

Characteristics of a respiratory syncytial virus persistently infected macrophage-like culture.

A persistently infected culture obtained from immortalized murine macrophage-like cells, which survived respiratory syncytial virus (RSV) infection at multiplicity of one, was established and characterized. The presence of RSV through the passages was confirmed and monitored by (a) detection of infectious virus by TCID(50)/ml, (b) defective particles by viral infectivity interference and buoyant density determinations, (c) cell surface antigen by indirect immunofluorescence and FACS, and (d) expression of a viral gene by RT-PCR. Moreover, cell morphology changes by comparison of macrophage area and perimeter were determined. A second culture was obtained by cell cloning out of this culture, and a third culture was established by superinfection with the original virus, in which 92-95% of the macrophages expressed viral antigen without cell destruction and released defective particles but low levels of infectious virus. Although the three cultures maintained the characteristics of persistently infected cells, concentrations of released infectious virus, defective particles, and percentages of cells bearing viral antigen varied. RSV persistently infected murine macrophage cultures provide an in vitro model to study viral-macrophage interaction and to allow the experimental use of a cell important in disseminating the infection. In addition, due to the wide array of cellular and humoral reagents in the mouse, studies on immunologic aspects of viral immunity are facilitated.