Effects of over-expression of TLR2 in transgenic goats on pathogen clearance and role of up-regulation of lysozyme secretion and infiltration of inflammatory cells

BMC Vet Res. 2012 Oct 22:8:196. doi: 10.1186/1746-6148-8-196.

Abstract

Background: Toll-like receptor 2 (TLR2) is important to host recognition of invading gram-positive microbes. In goats, these microbes can cause serious mastitis, anthrax, tetanus, and other problems. Transgenic goats constitutively over-expressing TLR2 in many tissues serve as a suitable model for the study of the role of TLR2 over-expression in bacterial clearance.

Results: Capra hircus TLR2 over-expression vector (p3S-LoxP-TLR2) was used to generate transgenic goats by egg microinjection. The integration efficiency was 8.57%. Real-time PCR and immunohistochemical results confirmed that the goats over-expressing the TLR2 gene (Tg) expressed more TLR2 than wild-type goats (WT). Monocyte-macrophages from the bloodstreams of transgenic goats were stimulated with synthetic bacterial lipoprotein (Pam3CSK4) and by the promotion of interleukin-6 (IL-6) and IL-10 expression in vitro. The oxidative damage was significantly reduced, and lysozyme (LZM) secretion was found to be up-regulated. Ear tissue samples from transgenic goats that had been stimulated with Pam3CSK4 via hypodermic injection showed that transgenic individuals can undergo the inflammation response very quickly.

Conclusions: Over-expression of TLR2 was found to decrease radical damage to host cells through low-level production of NO and MDA and to promote the clearance of invasive bacteria by up-regulating lysozyme secretion and filtration of inflammatory cells to the infected site.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Female
  • Goat Diseases / genetics
  • Goat Diseases / immunology
  • Goat Diseases / metabolism*
  • Goats
  • Histocytochemistry / veterinary
  • Leukocytes, Mononuclear / immunology
  • Leukocytes, Mononuclear / metabolism
  • Lipopolysaccharides / pharmacology
  • Malondialdehyde / metabolism
  • Muramidase / immunology
  • Muramidase / metabolism*
  • Nitric Oxide / metabolism
  • Oxidative Stress / physiology*
  • RNA / chemistry
  • RNA / genetics
  • Real-Time Polymerase Chain Reaction
  • Toll-Like Receptor 2 / biosynthesis*
  • Toll-Like Receptor 2 / genetics
  • Toll-Like Receptor 2 / immunology
  • Up-Regulation

Substances

  • Lipopolysaccharides
  • Toll-Like Receptor 2
  • Nitric Oxide
  • Malondialdehyde
  • RNA
  • Muramidase