Membrane orthologs of TLR5 of tongue sole Cynoglossus semilaevis: Expression patterns, signaling pathway and antibacterial property

Fish Shellfish Immunol. 2022 Jul:126:131-140. doi: 10.1016/j.fsi.2022.05.031. Epub 2022 May 23.

Abstract

Mammalian toll-like receptor 5 (TLR5) is crucial for recognizing bacterial flagellin and initiating the inflammatory signaling cascades via myeloid differentiation factor 88 (MyD88) signaling pathway, which plays vital roles in innate immune against pathogenic bacteria. Herein, we reported the signaling pathway and antibacterial property of tongue sole (Cynoglossus semilaevis) membrane forms of TLR5 (i.e. CsTLR5M1and CsTLR5M2). CsTLR5M1/M2 contain 936 and 885 amino acid residues respectively. CsTLR5M1 shares 86.7% overall sequence identities with CsTLR5M2. CsTLR5M1/M2 possess the same extracellular domain (ECD) and transmembrane domain (TMD), but the different toll-interleukin-1 receptor (TIR) domain. CsTLR5M1/M2 expression occurred constitutively in multiple tissues and regulated by bacterial stimulation. Recombinant CsTLR5M1/M2 (rCsTLR5M) could bind to flagellin and Gram-negative/positive bacteria, which could suppress bacterial growth. Stimulation of the CsTLR5M pathway by flagellin resulted in increased expression of MyD88-dependent signaling molecules and inflammatory cytokines. Blocking rCsTLR5M by antibody markedly reduced the phagocytosis and ROS production of peripheral blood leukocytes (PBLs), which in turn in vivo promoted the dissemination of bacteria. Overall, these observations add new insights into the signaling pathway and immune function of teleost TLR5M.

Keywords: Antibacterial immunity; Cynoglossus semilaevis; Expression pattern; MyD88-signaling pathway; Toll-like receptor 5.

MeSH terms

  • Animals
  • Anti-Bacterial Agents
  • Fish Diseases*
  • Fish Proteins
  • Flagellin / metabolism
  • Flagellin / pharmacology
  • Flatfishes*
  • Flounder* / metabolism
  • Gram-Negative Bacteria
  • Immunity, Innate / genetics
  • Mammals / metabolism
  • Myeloid Differentiation Factor 88 / genetics
  • Myeloid Differentiation Factor 88 / metabolism
  • Signal Transduction
  • Toll-Like Receptor 5 / genetics
  • Toll-Like Receptor 5 / metabolism

Substances

  • Anti-Bacterial Agents
  • Fish Proteins
  • Myeloid Differentiation Factor 88
  • Toll-Like Receptor 5
  • Flagellin