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Mol Immunol. 2007 May;44(12):3234-42. Epub 2007 Mar 6.

Synergy of CpG oligodeoxynucleotide and double-stranded RNA (poly I:C) on nitric oxide induction in chicken peripheral blood monocytes.

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Southern Plains Agricultural Research Center, USDA-ARS, College Station, 2881 F&B Road, TX 77845, USA. <>


Toll-like receptors (TLRs) recognize microbial components and initiate the innate immune responses that control microbial infections. We have investigated the innate immune response of chicken monocytes to ligands of TLR3 and TLR9, poly I:C (an analog of viral double-stranded RNA) and CpG-ODN (a CpG-motif containing oligodeoxydinucleotide) by measuring the induction of nitric oxide (NO) synthesis in chicken monocytes. Our results show that poly I:C and CpG-ODN synergized the induction of NO. When stimulated separately, CpG-ODN induced significant NO production in the chicken monocytes; whereas, poly I:C stimulated very little NO production. In combination, CpG-ODN and poly I:C induced significantly higher level of NO in chicken monocytes than either agonist alone. The addition of poly I:C prior to or simultaneously with CpG-ODN was required for the synergy. No synergistic effects on NO production were observed when monocytes were stimulated with combinations of CpG-ODN or poly I:C with other TLR agonists. Unlike chicken monocytes, cells of a chicken macrophage cell line, HD11, were readily stimulated to produce NO by both CpG-ODN and poly I:C with no synergism on NO induction when HD11 cells were stimulated by a combination of CpG-ODN and poly I:C. Using a pharmacological inhibitor, we also demonstrated that double-stranded RNA-dependent protein kinase (PKR) is indispensable for stimulation of NO production by CpG-ODN alone or in combination with poly I:C in both chicken peripheral blood monocytes and HD11 macrophage cells. Our results show that a combination of bacterial DNA and dsRNA induces an enhanced inflammatory immune response that has both antiviral and antibacterial activity in primary chicken monocytes.

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