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United States Department of Agriculture

Agricultural Research Service

Research Project: GENETIC AND BIOLOGICAL DETERMINANTS OF RESPIRATORY DISEASE SUSCEPTIBILITY Title: Innate immune responses of porcine macrophage cell line (Cdelts2+) to virus-associated virulence determinants

Authors
item Joseph, Jocelin -
item Chitko Mckown, Carol
item Kaushik, Radhey -

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: September 30, 2011
Publication Date: October 7, 2011
Citation: Joseph, J., Chitko-Mckown, C.G., Kaushik, R.S. 2011. Innate immune responses of porcine macrophage cell line (Cdelts2+) to virus-associated virulence determinants [abstract]. Prodeedings of the 71st Annual Meeting of the North Central Branch of the American Society of Microbiology, October 7-8, 2011, Des Moines, Iowa. Page 118. Abstract #34.

Technical Abstract: The goal of this study was to define the changes in the expression of immune genes in response to virus-associated virulence determinants. We stimulated a monocyte-derived porcine macrophage cell line (Cdelta2+) for 3 and 24h with Imiquimod, Poly IC and Poly IC with Lyovec. Cell lysates were processed for total RNA extraction and cDNA preparation. The gene expressions for TLRs, RIG-1, MDA-5, cytokines and defensins were quantified using real time RT-PCR. The expression of TLR 2 was downregulated by poly IC and poly IC with lyovec at 3h. TLR-3 gene was upregulated at 3 and 24h, while TLR-7 was upregulated at 3h with all three viral ligands. TLR-8 was upregulated by Imiquimod and Poly IC at 3h. Poly IC with Lyovec upregulated RIG-1 gene, but downregulated MDA-5 expression at 3h. All three viral ligands upregulated RIG-1 gene, but only poly IC upregulated MDA-5 gene at 24h.The expression of interferon-alpha was upregulated at 3h in response to all three viral ligands, while interferon-beta was upregulated at 3h in response to Imiquimod and Poly IC with Lyovec. IL-1alpha gene was upregulated at 3 and 24h, while IL-6 was upregulated at 3h by all three viral ligands. TNF-alpha expression was upregulated by Imiquimod at 3 and 24h. IL-12p35 and IL-8 were upregulated at 3h by all three viral ligands. Beta-defensin-1 and -2 were upregulated by Poly IC at 3h. Based on this study, we expect that the Cdelta2+ cell line may serve as a good model for studying the viral ligand interactions and virus pathogenesis.

Last Modified: 11/27/2014
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