Cytometry, titration and PCR results following infection of Antigen-presenting Cells with different strains of peste des petits ruminanst virus

[Lien vers la fiche source] This data corresponds to results of infections of monocyte-derived macrophages (MoMs) and monocyte-derived dendritic cells (MoDCs) from goats, sheep and cattle with two strains of peste des petits ruminants virus (PPRV). The two strains used in this study, Morocco 2008 (MA08) and Ivory Coast 1989 (IC89), demonstrated different levels of virulence in certain goat breeds. Strain MA08 belongs to lineage IV of PPRV and was isolated from an Alpine goat that was severely affected during the 2008 outbreak in Morocco. The IC89 strain belongs to lineage I. It was isolated from a goat in Ivory Coast in 1989 and is considered to be a mildly virulent strain, inducing a mild disease in various goat breeds, although it is used as a virulent strain for the study of immune response in sheep. Based on representative known virulence to goats of the two strains of PPRV, low for IC89 and high for MA08, the aim of this study was to assess the comparative in vitro replication of these PPRV strains in both MoMs and MoDCs of goats, sheep and cattle. The research question was to evaluate whether any difference could be observed between these APCs in relation to PPRV strain virulence. To determine the infectivity of the two PPRV strains, IC89 and MA08, for macrophages and dendritic cells, circulating monocytes from the blood of the different animal species studied were differentiated into dendritic cells (MoDCs) and macrophages (MoMs). These two cell types were infected with each of the viral strains studied, IC89 and MA08, for 48 hours at a multiplicity of infection (MOI) of 0.1. After observing low or non-detectable viral titers in supernatants of goat MoMs infected for 48 hours with the IC89 strain, infection kinetics were studied to see if replication was somewhat delayed for this low virulent strain. Goat MoMs were infected for 24 h, 48 h, 72 h and 96 h, at an MOI of 0.1. Test were done on individual biological replicates from a minimum of three animals per species and per condition. Effects of virus infection on the APCs were evaluated with three different type of tests: 1) Flow cytometry after intracellular labelling with an antibody targeting the virus nucleoprotein (anti-NPPRV) coupled to a fluorochromeflow cytometry. Results obtained correspond to percentage of cells showing fuorescence (= infected cells). 2) Virus titer in cultures of SLAM-expressing Vero cells (VDS) where post-infection supernatants were added at dilutions from 10-1 to 10-7 with a minimum of four replicates per dilution. Results are presented as TCID50/mL . 3) RT-qPCR targeting a portion of the PPRV genome to detect presence of virus genetic material in post-infection supernatants. Results are presented as Cq values. This data was analysed using generalised linear mixed models. Species, cell type, and viral strain were included as fixed effects, and a nested random effect (1| ID/Sample) was included to account for repeated measurements within individuals, where ID represents animal identitfy and Sample /or represents the experimental sample. This dataset includes a text file describing the data files included in this dataset and three data files in CSV format.