Excretory/secretory proteins of adult Toxocara canis induce changes in the expression of proteins involved in the NOD1-RIP2-NF-κB pathway and modulate cytokine production in mouse macrophages

Exp Parasitol. 2021 Oct:229:108152. doi: 10.1016/j.exppara.2021.108152. Epub 2021 Aug 19.

Abstract

Dog roundworm (Toxocara canis) is the major causative agent of toxocarosis, a parasitic disease of both veterinary and medical importance. Knowledge gaps in fundamental and applied aspects hinder the control of this important zoonotic disease. To have a better understanding of Toxocara infection and host immune responses, mouse macrophages were exposed to excretory/secretory (ES) proteins released by adult worms of T. canis in vitro. The messenger RNA transcription and protein expression of nucleotide-binding oligomerization domain-containing protein 1 (NOD1), receptor interacting protein 2 (RIP2) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) in macrophages were analysed using quantitative real-time PCR (qRT-PCR) and Western blot. The levels of tumour necrosis factor alpha (TNF-ɑ), interleukin-1 beta (IL-1β) and IL-6 released by the stimulated macrophages were analysed using enzyme-linked immunosorbent assay. It was found that 20 μg/mL ES proteins of adult T. canis induced the expression of NOD1, RIP2 and NF-κB in mouse macrophages at both transcriptional and translational levels after 9 h of incubation in vitro. Incubation with 20 μg/mL ES proteins also modulated the production of pro-inflammatory cytokines TNF-ɑ, IL-1β and IL-6 by the macrophages. Taken together, ES proteins of adult T. canis appeared to be able to affect the macrophage NOD1-RIP2-NF-κB signalling pathway, which might play a role in regulating the production of proinflammatory cytokines. Further investigation of these aspects should lead to a better understanding of immune recognition of and modulation by Toxocara canis in host animals.

Keywords: Cytokine production; Excretory/secretory proteins; Mouse macrophage; NOD1-RIP2-NF-κB pathway; Toxocara canis.

MeSH terms

  • Animals
  • Blotting, Western
  • Cell Survival
  • Cytokines / biosynthesis*
  • Cytokines / metabolism
  • Dogs
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Helminth Proteins / metabolism*
  • Helminth Proteins / pharmacology
  • Interleukin-1beta / biosynthesis
  • Interleukin-1beta / metabolism
  • Interleukin-6 / biosynthesis
  • Interleukin-6 / metabolism
  • Macrophages, Peritoneal / drug effects
  • Macrophages, Peritoneal / metabolism*
  • Macrophages, Peritoneal / parasitology
  • Mice
  • Mice, Inbred C57BL
  • NF-kappa B / metabolism
  • Nod1 Signaling Adaptor Protein / metabolism
  • Receptor-Interacting Protein Serine-Threonine Kinase 2 / metabolism
  • Toxocara canis / chemistry
  • Toxocara canis / metabolism*
  • Tumor Necrosis Factor-alpha / biosynthesis
  • Tumor Necrosis Factor-alpha / metabolism

Substances

  • Cytokines
  • Helminth Proteins
  • Interleukin-1beta
  • Interleukin-6
  • NF-kappa B
  • Nod1 Signaling Adaptor Protein
  • Tumor Necrosis Factor-alpha
  • Receptor-Interacting Protein Serine-Threonine Kinase 2