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Background  NLRC5 is a member of the CARD domain containing, nucleotide-binding oligomerization (NOD)-like receptor (NLR) family, which recognizes pathogen-associated molecular patterns (PAMPs) and initiates an innate immune response leading to inflammation and/or cell death. However, the specific role of  NLRC5  as a modulator of the inflammatory immune response remains controversial. It has been reported to be a mediator of type I IFNs, NF-kB, and  MHC class I  gene. But no study on  NLRC5  function has been reported to date in chickens. In the current study, we investigated the role of  NLRC5  in the regulation of  IFNA ,  IFNB ,  IL-6 , and  MHC class I  in the chicken HD11 macrophage cell line, by using RNAi technology. HD11 cells were transfected with one of five siRNAs (s1, s2, s3, negative-siRNA, or a mixture of s1, s2, s3-siRNAs). After 24 hours, cells were exposed to LPS or poly (I:C) or a vehicle control. Gene expression of  NLRC5 ,  IFNA ,  IFNB ,  IL-6 , and  MHC class I  at 2, 4, 6, and 8 hours post stimulation (hps) was quantified by qPCR.    Results  The expression of  NLRC5 ,  IFNA ,  IFNB , and  IL-6  genes in negative irrelevant transfection controls was up-regulated at 2 hps after LPS treatment compared to the vehicle controls. S3-siRNA effectively knocked down  NLRC5  expression at 4 hps, and the expression of  IFNA  and  IFNB  (but not  IL-6  and  MHC class I ) was also down-regulated at 4 hps in s3-siRNA transfected cells, compared to negative irrelevant transfection controls. Stimulation by LPS appeared to relatively restore the decrease in  NLRC5 ,  IFNA , and  IFNB  expression, but the difference is not significant.    Conclusions  Functional characterization of chicken  NLRC5  in an  in vitro  system demonstrated its importance in regulating intracellular molecules involved in inflammatory response. The knockdown of  NLRC5  expression negatively mediates gene expression of  IFNA  and  IFNB  in the chicken HD11 cell line; therefore,  NLRC5  likely has a role in positive regulation of  IFNA  and  IFNB  expression. No direct relationship was found between  NLRC5  knockdown and  IL-6  and  MHC class I  expression. Future studies will further clarify the roles of  NLRC5  and other NLRs in infectious diseases of chickens and may increase the efficacy of antiviral vaccine design.

NLRC5 knockdown in chicken macrophages alters response to LPS and poly (I:C) stimulation