Submitted to: International Positive Strand RNA Virus Symposium
Publication Type: Abstract Only
Publication Acceptance Date: 3/11/2010
Publication Date: 5/17/2010
Citation: Miller, L.C., Harhay, G.P., Lager, K.M., Neill, J.D., Kehrli, Jr., M.E. 2010. In-Depth Global Analysis of Transcript Abundance Levels Following Infection with Porcine Reproductive and Respiratory Syndrome Virus [abstract]. In: Proceedings of the 9th International Positive Strand RNA Virus Symposium, May 17-21, 2010, Atlanta, Georgia. p. 54.
Technical Abstract: Porcine reproductive and respiratory syndrome virus (PRRSV) is a major pathogen of swine worldwide and causes considerable economic loss. Infection of the primary target cells, porcine alveolar macrophages (PAMs), by PRRSV causes significant changes in their function by mechanisms that are not understood. Identifying specific pathways that associate with variation in PRRSV replication and macrophage function may lead to novel gene targets for the control of PRRSV infection. Serial Analysis of Gene Expression (SAGE) libraries were constructed from in vitro mock-infected and PRRSV strain VR-2332-infected PAMs at 0, 6, 12, 16 and 24 hours post-infection. Each SAGE library was sequenced to obtain >95,000 tags per time point. Tags were mapped to transcripts and genes by exact regular expression matching to sequences in GenBank, Harvard Gene Index, and the Pig Expression Database (Japan). Examination of the SAGE data indicated that there were changes in transcript abundance occurring in the PRRSV-infected PAM over time post-infection. More than 590 unique tags with significantly altered transcript abundance levels were identified (p<0.01 with Bonferroni correction). The validity and kinetics of transcript abundance of SAGE identified genes were confirmed using real-time RT-PCR. The most striking finding was that the transcript abundance of most of the identified genes involved in the innate immune response (including IL-8, CCL4, and IL-1ß, genes whose transcript abundances are typically altered in response to other pathogens or insults) showed no or very little change at any time point following infection.