Submitted to: Research Workers in Animal Diseases Conference Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: 9/5/2008
Publication Date: 12/5/2008
Citation: Wysocki, M., Steibel, J.P., Mccaw, M., Kuhar, D.J., Ernst, C.W., Lunney, J.K. 2008. Uncovering genetic components involved in early regulatory immune response during PRRSV infection [abstract]. Conference Research Workers in Animal Diseases Conference. Paper No. 125.
Technical Abstract: Our goal is to identify the most significant pathways involved in early immune responses during porcine reproductive and respiratory syndrome virus (PRRSV) infection as compared to protective vaccination. For this experiment PRRSV-naïve animals were divided into four groups: (1) pigs infected with Acute-type PRRSV MNW2B; (2) pigs infected with Acute-type PRRSV NC Powell; (3) pigs vaccinated with a commercial modified-live PRRSV vaccine (Ingelvac ATP®) and (4) control pigs. Tissues [tracheobronchial lymph nodes (TBLN), cranial and caudal lung lobes, and tonsils] were collected between 3-6 days post infection/vaccination. Total RNA was prepared, labeled with Alexa Fluor® 555 and 647 dyes (Invitrogen) and hybridized to the Swine Protein-Annotated Oligonucleotide Microarray (20,400 oligos, www.pigoligoarray.org). A microarray loop design was used to test for changes in gene expression between all four groups and differences due to day post infection within the PRRSV infected groups. Results obtained for cranial lung tissues affirmed 923, 619 and 747 putatively differentially expressed genes for comparisons of different groups: (1) vaccinated versus control, (2) vaccinated versus infected, and (3) control versus infected, respectively. Final statistical and pathway analyzes are underway for lung, TBLN and tonsils to identify the biological functions and regulatory pathways that are most significant and genes whose expression is altered by the two pathologic acute PRRSV infections as compared to the protective vaccination. As expected, pathways involving interferons and other cytokines, as well as chemokines, have been identified as critical for differentiating infected from vaccinated pigs. Subsequent experiments will identify candidate genes and carry out real time RT- PCR to confirm their differential expression. Supported by USDA ARS and CSREES PRRS CAP funds.