Recombinant adenovirus vector bearing antisense macrophage migration inhibitory factor cDNA prevents acute lipopolysaccharide-induced liver failure in mice.

Abstract

Macrophage migration inhibitory factor (MIF) is a pleiotropic cytokine involved in delayed hypersensitivity and cellular immunity. MIF also acts as a proinflammatory cytokine and counterregulates the anti-inflammatory effects of glucocorticoids. Exogenous gene transfer mediated by adenovirus is useful to study a particular molecular function as well as to develop gene therapy strategies. A recombinant adenovirus containing sense and antisense murine MIF (mMIF) cDNA inserts was constructed using a cosmid-terminal protein complex method. The sense mMIF adenovirus (AxCA-mMIFS) efficiently induced mMIF in COS-7 cells that endogenously lack mMIF in a dose-dependent manner. In contrast, the antisense mMIF adenovirus (AxCA-mMIFAS) inhibited the expression of mMIF in NIH3T3 cells in a dose-dependent manner. To assess the pathophysiologic role of MIF in acute liver failure, we induced acute onset of liver damage in mice (male Jcl:ICR) by a combined treatment of Bacille Calmette-Guerin (BCG) and lipopolysaccharide (LPS). mMIF level in the liver of mice infected with AxCA-mMIFAS showed a significant reduction in MIF production in response to BCG-LPS compared with mice treated without viral infection and with AxCA-mMIFS. In addition, the immunohistochemical staining demonstrated that F4/80 antigen on macrophage was enhanced in liver infected with AxCA-mMIFS but reduced in liver infected with AxCA-mMIFAS. The staining intensity is correlated with the mMIF antigen level in liver tissue. The survival rate of mice infected with AxCA-mMIFAS was significantly higher than that of mice treated with PBS and infected with AxCA-LacZ in BCG-LPS. These results suggest that inhibition of MIF production, using recombinant adenovirus bearing the antisense MIF gene, reduced the mortality rate in BCG-LPS-induced liver failure in mice. This finding might aid in the further development of gene therapy targeting MIF.

• Authors: Takayuki Iwaki, Motoi Sugimura, Jun Nishihira, Toshiki Matsuura, Takao Kobayashi, Naohiro Kanayama
• Journal: Laboratory investigation; a journal of technical methods and pathology (Lab Invest) Vol. 83 Issue 4 Pg. 561-70 (Apr 2003) ISSN: 0023-6837 [Print] United States
• PMID: 12695559 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• BCG Vaccine
• DNA, Complementary
• Lipopolysaccharides
• Macrophage Migration-Inhibitory Factors
• Oligonucleotides, Antisense
• Recombinant Fusion Proteins

Topics

• 3T3 Cells
• Adenoviridae (genetics)
• Animals
• BCG Vaccine (administration & dosage)
• COS Cells
• DNA, Complementary (genetics)
• Disease Models, Animal
• Escherichia coli (immunology)
• Gene Transfer Techniques
• Genetic Vectors
• Lipopolysaccharides (toxicity)
• Liver (drug effects, metabolism, pathology)
• Liver Failure (chemically induced, mortality, pathology, prevention & control)
• Macrophage Migration-Inhibitory Factors (genetics, metabolism)
• Male
• Mice
• Mice, Inbred ICR
• Oligonucleotides, Antisense (therapeutic use)
• Recombinant Fusion Proteins
• Specific Pathogen-Free Organisms
• Survival Rate
• Transfection