E-GEOD-40644 - Transcriptional analysis of Bos taurus and AlHV-1 in MDBK infected cells and in lymph nodes of infected calves
Released on 21 March 2013, last updated on 2 April 2013
Wildebeests carry asymptomatically Alcelaphine herpesvirus 1 (AlHV-1), a γ-herpesvirus inducing a lethal lymphoproliferative disease named malignant catarrhal fever (MCF) in a number of susceptible species of the Artiodactyla order, including cattle. The local population welfare in eastern Africa is directly endangered by the important but underestimated impact of this disease on their livelihood. Although AlHV-1 genomic DNA is detected in abundance in tissues during MCF, no infectious viral particles and very low viral protein expression levels are observed. This suggests that AlHV-1 might be latent during MCF. Here, we studied the implication of AlHV-1 latency during MCF. We first examined the expression of poly-adenylated RNA from infected (multiplicity of infection, moi = 0.01) MDBK cells at 72h pi. This late time point was chosen as we expect the majority of viral genes to be expressed. The expression was obtained from two-color dye-swap analyses of 4 independent biological repeats. To determine cellular and viral gene expression during MCF, we extracted RNA from the inguinal LN (iLN) of each calf for analysis on a custom designed array. The arbitrary choice of the iLN as the selected tissue was based on the fact that AlHV-1 viral genomic load are the highest in the LN. Cellular and viral RNA transcription profiles were analyzed with two-color dye-swap analyses of 4 independent biological repeats. Cellular and viral gene expression were analysed in Mock- and AlHV-1-infected MDBK cells (in vitro) as well as in the inguinal lymphnodes of Mock- and AlHV-1-infected calves (in vivo). Each experiment (in vitro and in vivo) was carried out with 4 biological replicates for each conditon (mock- and AlHV-1-infected). The 4 samples for each experiment were hybridized in a one-to-one dye-swap design without pooling the Mock-infected samples, and yielding 8 arrays per experiment.
transcription profiling by array