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Screening Bioactive Nanoparticles in Phagocytic Immune Cells for Inhibitors of Toll-like Receptor Signaling
Published on: July 26, 2017
Scallop toll-like receptor engages in complex interactions with diverse MyD88 proteins and orchestrates host innate
Jilv Ma1, Fengchen Liu2, Zihao Zhang3
1School of Fisheries, Ludong University, Yantai, China; College of Animal Science and Technology, Guangxi University, Nanning, China.
Abstract:
Toll-like receptor (TLR) family proteins act as pattern recognition receptors with pivotal roles in immune defense; however, the function of TLRs in marine mollusks requires further investigation. In this study, we identified a novel TLR gene (CfTLR-Like-1 [CfTLR-L1]) in Chlamys farreri and analyzed its immune functions. CfTLR-L1 has a coding sequence of 1785 bp and encodes 594 amino acids. It includes typical protein domains; the amino acid sequences of the protein domains are conserved. Phylogenetic analyses demonstrated a close evolutionary relationship among proteins in the mollusk TLR subfamily. CfTLR-L1 is predominantly located on the cell membrane. CfTLR-L1 mRNA is expressed in all tissues, particularly in the hepatopancreas and mantle. Pathogenic stressors, including lipopolysaccharide, peptidoglycan, polyinosinic-polycytidylic acid, Vibrio anguillarum, Vibrio alginolyticus, Vibrio splendidus, and acute viral necrosis virus, significantly increase the levels of CfTLR-L1 mRNA transcripts. The Toll/interleukin 1 receptor (TIR) domain of CfTLR-L1 interacts with the MyD88 family proteins of scallops (including CfMyD88-1, CfMyD88-3, and CfMyD88-4), implying that multiple unique TLR signal transduction mechanisms exist in marine mollusks. CfTLR-L1-TIR interacts with CfMyD88-1-P3, which contains only the TIR structural domain. Overexpression of CfTLR-L1-TIR in HEK293T cells activated reporter genes, including IFNβ, IFNγ, and STAT3. Similarly, overexpression of CfTLR-L1 significantly affected mitogen-activated protein kinase phosphorylation, specifically of p38 and c-Jun N-terminal kinase. Our findings expand the knowledge of the role of TLR in mollusk immunity and may provide evidence for selecting disease-resistant scallops for cultivation.
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