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Toll-like receptors (TLRs) mediated signaling plays a vital role in activating innate and adaptive immunity. Although TLR mediated signaling has been comprehensively investigated in mammalian species, the mechanisms underlying TLR signaling in molluscs remain obscure. In the present study, a novel TLR isoform namely McTLR-like1 was identified in the thick shell mussel Mytilus coruscus. McTLR-like1 was highly expressed in molluscan immune-related tissues, and its transcriptional levels in hemocytes were significantly increased when challenged by V. alginolyticus. McTLR-like1 activated nuclear factor κB (NF-κB) and strengthened the transcription and phosphorylation of NF-κB subunit P65 in mammalian cells. Upon the silencing of McTLR-like1, the mRNA expression levels of pro-inflammatory cytokines were down-regulated, and the animals exhibited higher levels of resistance when challenged with V. alginolyticus. McMyD88a mRNA expression was also downregulated alongside McTLR-like1. Furthermore, GST-pull down assays revealed a visible affinity between McTLR-like1 and McMyD88a. Collectively, these results demonstrated that the newly identified gene affiliated to the molluscan TLR family and plays a role in the TLR-mediated activation of inflammatory response via its affinity with MyD88. The present study enhances our knowledge of TLR signaling mechanisms in molluscs and provides new insights into the evolution of TLRs., (Copyright © 2022 Elsevier Ltd. All rights reserved.)