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Article
Reduced Nitric Oxide Production and iNOS mRNA Expression in IFN-γ-Stimulated Chicken Macrophages Transfected with iNOS siRNAs
Veterinary Immunology and Immunopathology
  • Jennifer H. Cheeseman, Iowa State University
  • Hyun S. Lillehoj, United States Department of Agriculture
  • Susan J. Lamont, Iowa State University
Document Type
Article
Publication Version
Published Version
Publication Date
10-1-2008
DOI
10.1016/j.vetimm.2008.05.015
Abstract

Utilizing RNA interference technology with siRNA in the HD11 macrophage cell line, we determined how the inhibition or knock-down of the iNOS (inducible nitric oxide synthase) gene affected IFN-γ-induced macrophage production of nitric oxide (NO) and mRNA expression of genes involved in this biological pathway in the chicken. Chicken macrophages produce NO when stimulated with recombinant chicken IFN-γ, however, when transfected with iNOS siRNAs, the production of NO is significantly decreased. We observed a 14–28% reduction in NO production by IFN-γ-stimulated HD11 cells at 48 h after initial siRNA transfection compared to non-transfected IFN-γ-stimulated macrophages. Significant knock-down of iNOS mRNA expression (15 to 50-fold lower) was observed for each of four iNOS siRNAs, when compared to non-transfected IFN-γ-stimulated macrophages and to those treated with a negative control siRNA. The IFN-γ-stimulated chicken macrophages transfected with iNOS siRNAs did not show altered levels of mRNA expression for genes involved in IFN-γ signaling and iNOS pathways (IL-1β, IL-6, IFN-γ, TGF-β4, or SOCS-3) suggesting that the observed decrease in NO production is a direct result of siRNA mediated knock-down of iNOS, rather than IFN-γ-induced changes in the other genes tested.

Comments

This article is from Veterinary Immunology and Immunopathology 125 (2008): 375, doi:10.1016/j.vetimm.2008.05.015.

Rights
Works produced by employees of the U.S. Government as part of their official duties are not copyrighted within the U.S. The content of this document is not copyrighted.
Language
en
File Format
application/pdf
Citation Information
Jennifer H. Cheeseman, Hyun S. Lillehoj and Susan J. Lamont. "Reduced Nitric Oxide Production and iNOS mRNA Expression in IFN-γ-Stimulated Chicken Macrophages Transfected with iNOS siRNAs" Veterinary Immunology and Immunopathology Vol. 125 Iss. 3-4 (2008) p. 375 - 380
Available at: http://works.bepress.com/susan_lamont/70/