Screening of Optimal CpG-Oligodeoxynucleotide for Anti-Inflammatory Responses in the Avian Macrophage Cell Line HD11

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CpG-oligodeoxynucleotides (CpG-ODNs ) have been shown to possess immunostimulatory features in both mammals and birds. However, compared to their proinflammatory effects,little is known about the anti-inflammatory responses triggered by CpG-ODN in avian cells. Hence, in this study, the anti-inflammatory response in the chicken macrophage cell line HD11 was characterized under stimulation with five types of CpG-ODNs: CpG-A₁₅₈₅, CpG-A_D35, CpG-B₁₅₅₅,CpG-B_K3, and CpG-C₂₃₉₅. Single-stimulus of CpG-B₁₅₅₅, CpG-B_K3, or CpG-C₂₃₉₅ induced interleukin (IL)-10 expression without causing cell injury. The effects of pretreatment with CpG-ODNs before subsequent lipopolysaccharide stimulation were also evaluated. Interestingly, pretreatment with only CpG-C₂₃₉₅ resulted in high expression levels of IL-10 mRNA in the presence of lipopolysaccharide. Finally, gene expression analysis of inflammation-related cytokines and receptors revealed that pre-treatment with CpG-C₂₃₉₅ significantly reduced the mRNA expression of tumor necrosis factor-α,IL-1β, IL-6, and Toll-like receptor 4.Overall, these results shed light on the anti-inflammatory responses triggered by CpG-C₂₃₉₅ stimulation through a comparative analysis of five types of CpG-ODNs in chicken macrophages. These results also offer insights into the use of CpG-ODNs to suppress the expression of proinflammatory cytokines, which may be valuable in the prevention of avian infectious diseases in the poultry industry.