Identification and characterization of the toll-like receptor 8 gene in the Chinese raccoon dog (Nyctereutes procyonoides)

文献类型： 外文期刊

第一作者： Yang, Yong

作者： Yang, Yong;Yang, Yong;Tong, Mingwei;Yi, Li;Cheng, Yuening;Zhang, Miao;Cao, Zhigang;Wang, Jianke;Lin, Peng;Cheng, Shipeng

作者机构：

关键词： TLR8;Nyctereutes procyonoides;Sequence analysis;Gene expression

期刊名称：IMMUNOLOGY LETTERS （ 影响因子：3.685； 五年影响因子：3.54 ）

ISSN：

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收录情况： SCI

摘要： TLR8 is an important sensor of single-stranded RNA (ssRNA) from the viral genome and plays an essential role in innate antiviral responses via the recognition of conserved viral molecular patterns. In this report, TLR8 in the Chinese raccoon dog was characterized and analyzed for the first time. The fulllength sequence of raccoon dog TLR8 (RdTLR8) cDNA was cloned by rapid amplification of cDNA ends (RACE) and is 3191bp with a 3117-bp open reading frame (ORF) encoding 1038 amino acids. The putative protein exhibits typical features of the TLR families, with 19 leucine-rich repeats (LRRs) in the extracellular domain and a cytoplasmic TIR domain. Comparative analyses of the RdTLR8 amino acid sequence indicated a 73.6-99.4% sequence identity with dog, horse, pig, sheep, cattle, human and mouse TLR8. Phylogenetic analysis grouped 71 mammalian TLR proteins into five sub-families, wherein RdTLR8 was clustered into a monophyletic TLR8 Glade in the TLR9 family, which was completely coincident with the evolutionary relationship among mammals. Quantitative real-time PCR analysis revealed extensive expression of RdTLR8 in tissues from healthy Chinese raccoon dogs with the highest expression in the peripheral blood mononuclear cells (PBMCs) and the lowest expression in the skeletal muscle. HEK293 cells cotransfected with a RdTLR8 expression plasmid and an NF-kappa B-luciferase reporter plasmid significantly responded to the agonist 3M-002, indicating a functional TLR8 homolog. In addition, raccoon dog PBMCs exposed to the canine distemper virus (CDV) wild strain CDV-PS and the TLR8 agonist 3M-002 showed significant upregulation of RdTLR8 mRNA and proinflammatory cytokines TNF-alpha and IFN-alpha, suggesting that RdTLR8 might play an important role in the immune response to viral infections in the Chinese raccoon dog. (C) 2016 European Federation of Immunological Societies. Published by Elsevier B.V. All rights reserved.

分类号： R392

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