Abstract

Oligodeoxynucleotides containing unmethylated CpG motifs (CpG ODN) mimic the immunostimulatory activity of microbial DNA by interacting with Toll-like receptor 9 (TLR9) to activate both the innate and adaptive immune responses in different species. However, few studies have been published to compare the effects of CpG ODN on different pig breeds. Therefore, in this study, whole blood gene expression profiles of DPL and Landrace pigs treated with CpG ODN were studied using RNA-seq technology. Five Hundred differentially expressed genes (DEGs) were identified between the two breeds. DPL pigs had significantly higher number of immune-relevant DEGs than the Landrace pigs after CpG ODN treatment. Pathway analysis showed that cytokine-cytokine receptor interaction and chemokine signaling pathway were the major enriched pathways of the immune-relevant DEGs. Further in vitro experiments showed that PBMCs of the DPL pigs had significantly higher levels of TLR9 mRNA than those of the Landrace pigs, both before and after CpG ODN stimulation. Cytokine and chemokine induction in the PBMCs of both breeds were also measured after CpG ODN stimulation. Our data showed that mRNA levels of cytokines (IFNα, IL8, IL12 p40) and chemokines (CXCL9, CXCL13) were significantly higher in the PBMCs of the DPL pigs than those of the Landrace pigs. Taken together, our data provide new information regarding the pig breed difference in response to CpG ODN stimulation and that higher levels of TLR9 mRNA in DPL pigs may be a major contributor for disease resistance.

Highlights

  • In the pig production industry, infectious diseases caused by viral and bacterial pathogens seriously influence animal welfare, product quality and economics around the world (Meng, 2012)

  • Based on the RNA-seq results, we further studied the mRNA expression of Toll-like receptor 9 (TLR9) and cytokine/chemokine in the peripheral blood mononuclear cells (PBMCs) of DPL and Landrace pigs after C CpG ODN 2429 stimulation in vitro

  • The large number of differentially expressed genes (DEGs) identified suggest that the DPL and Landrace pigs responded to CpG ODN stimulation quite differently

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Summary

Introduction

In the pig production industry, infectious diseases caused by viral and bacterial pathogens seriously influence animal welfare, product quality and economics around the world (Meng, 2012). Breed is one of the most important factors that directly influence resistance or susceptibility to various infectious diseases (Reiner et al, 2002; Opriessnig et al, 2006; Lunney and Chen, 2010). Chinese indigenous pig breeds are generally better at disease resistance than foreign lean-type pig breeds (Yang, 2013). Dapulian (DPL) pigs are an indigenous breed of China, which mainly distribute in Shandong Province, China. Compared to modern commercial pig breeds, DPL pigs exhibit stronger resistance to diseases such as porcine reproductive and respiratory syndrome (PRRS) (Jiang et al, 2013; Xing et al, 2014). Since DPL and Landrace pigs were evolved under different environment

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