Cloning and characterization of giant panda (Ailuropoda melanoleuca) IL-18 binding protein

Res Vet Sci. 2016 Jun:106:170-2. doi: 10.1016/j.rvsc.2016.04.004. Epub 2016 Apr 27.

Abstract

The giant panda (Ailuropoda melanoleuca) is an endangered species. Interleukin-18 (IL-18) plays an important role in the innate and adaptive immune responses by inducing IFN-γ. IL-18 has been implicated in the pathogenesis of various diseases. IL-18 binding protein (IL-18BP) is an intrinsic inhibitor of IL-18 that possesses higher affinity to IL-18. In this study, we cloned and characterized IL-18BP in giant panda (AmIL-18BP) from the spleen. The amino acid sequence of giant panda IL-18BP ORF shared about 65% identities with other species. To evaluate the effects of AmIL-18BP on the immune responses, we expressed the recombinant AmIL-18BP in Escherichia coli BL21 (DE3).The fusing protein PET-AmIL-18BP was purified by nickel affinity column chromatography. The biological function of purified PET-AmIL-18BP was determined on mice splenocyte by qRT-PCR. The results showed that AmIL-18BP was functional and could significantly reduce IFN-γ production in murine splenocytes. These results will facilitate the study of protecting giant panda on etiology and immunology.

Keywords: Bioactivity analysis; Giant panda; IL-18 binding protein; Interleukin-18.

MeSH terms

  • Animals
  • Cloning, Molecular
  • DNA, Complementary / genetics
  • DNA, Complementary / metabolism
  • Intercellular Signaling Peptides and Proteins / chemistry
  • Intercellular Signaling Peptides and Proteins / genetics*
  • Intercellular Signaling Peptides and Proteins / metabolism
  • Interferon-gamma / genetics
  • Interferon-gamma / metabolism
  • Mice
  • Sequence Analysis, Protein / veterinary
  • Spleen / immunology
  • Ursidae / genetics*
  • Ursidae / immunology

Substances

  • DNA, Complementary
  • IFNG protein, mouse
  • Intercellular Signaling Peptides and Proteins
  • interleukin-18 binding protein
  • Interferon-gamma