Expression and characterization of recombinant interferon gamma (IFN-gamma) from the nine-banded armadillo (Dasypus novemcinctus) and its effect on Mycobacterium leprae-infected macrophages

Cytokine. 2008 Aug;43(2):124-31. doi: 10.1016/j.cyto.2008.04.014. Epub 2008 Jun 16.

Abstract

Armadillos (Dasypus novemcinctus) manifest the full histopathological spectrum of leprosy, and are hosts of choice for in vivo propagation of Mycobacterium leprae. Though potentially useful as a model of leprosy pathogenesis, few armadillo-specific reagents exist. We have identified a region of high homology to the interferon gamma (IFN-gamma) of other mammals within the recently published armadillo whole genomic sequence. cDNA was made from ConA-stimulated armadillo peripheral blood mononuclear cells (PBMC), amplified, and cloned into a pET expression vector for transformation and over-expression in Escherichia coli. The recombinant protein (rDnIFN-gamma) was characterized by western blot and its biological function confirmed with bioassays including intracellular killing of Toxoplasma gondii and induction of indoleamine 2, 3-dioxygenase activity. In using rIFN-gamma to activate macrophages from mice, humans or armadillos, similar to humans, rIFN-gamma-activated armadillo MPhi did not produce nitrite and or inhibit the viability of M. leprae in vitro. Conversely, murine rIFN-gamma-activated mouse MPhi produced high levels of nitrite and killed intracellular M. leprae in vitro. These data indicate that the response of armadillo MPhi to rDnIFN-gamma is similar to that which occurs in humans, and demonstrates a potentially important value of the armadillo as a model in leprosy research.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies / immunology
  • Armadillos / genetics
  • Armadillos / metabolism*
  • Base Sequence
  • Cell Proliferation / drug effects
  • DNA, Complementary / genetics
  • Gene Expression*
  • Humans
  • Indoleamine-Pyrrole 2,3,-Dioxygenase / metabolism
  • Interferon-gamma / chemistry
  • Interferon-gamma / genetics
  • Interferon-gamma / metabolism*
  • Interferon-gamma / pharmacology*
  • Macrophages / cytology
  • Macrophages / drug effects*
  • Molecular Sequence Data
  • Mycobacterium leprae / physiology*
  • Nitrites / metabolism
  • Protein Isoforms / chemistry
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • Protein Isoforms / pharmacology
  • Recombinant Proteins
  • Sequence Alignment
  • Sequence Homology, Amino Acid

Substances

  • Antibodies
  • DNA, Complementary
  • Indoleamine-Pyrrole 2,3,-Dioxygenase
  • Nitrites
  • Protein Isoforms
  • Recombinant Proteins
  • Interferon-gamma

Associated data

  • GENBANK/DQ094083