Regulation of ornithine decarboxylase in the kidney of autoimmune mice with the lpr gene.
The lymphoproliferative lpr gene confers a lupus-like disease with lymphadenopathy, antinuclear antibody production, and glomerulonephritis in MRL-lpr/lpr mice. Upregulation of ornithine decarboxylase (ODC) activity and polyamine levels have been observed in the kidney and lymphoid organs of this strain. Inhibition of ODC with 0.5-1.5% (w/v) difluoromethylornithine (DFMO) in drinking water prolonged life-span and ameliorated renal disease. Glomerulonephritis is a major cause of morbidity and mortality in human and murine lupus. In order to elucidate the mechanism(s) of ODC regulation in lupus nephritis, we characterized ODC at the protein and mRNA levels in 3 strains of autoimmune mice with the lpr genetic background (MRL-lpr/lpr, C3H-lpr/lpr and C57BL/6J-lpr/lpr) using Western blotting, enzyme kinetics, turnover rate measurements, Northern blot hybridization, and reverse transcription-polymerase chain reaction (RT-PCR). Normal BALB/c mice were used as a control. We found that ODC activity in the kidney of lpr strains was 4- to 6-fold higher than that of BALB/c mice. The intensity of the major ODC protein band at 54 kD in Western blot was 4-fold higher in MRL-lpr/lpr and C3H-lpr/lpr kidney compared to that of BALB/c kidney. Putrescine levels were 2- to 4-fold higher in kidney of lpr strains than that of BALB/c and DFMO-treated MRL-lpr/lpr mice. DFMO treatment significantly reduced ODC activity and polyamine levels. The half-life of ODC enzyme in MRL-lpr/lpr, C3H-lpr/lpr, B6-lpr/lpr and BALB/c mouse kidneys was 15, 5, 8 and 23 min, respectively. There was no significant difference in the Km values of different strains, whereas Vmax values differed significantly. There was no difference in the level of SAMDC, another enzyme involved in the polyamine biosynthetic pathway, in various strain. Steady-state levels of ODC mRNA were lower in lpr strains compared to that of BALB/c mouse. Our results suggest that the basis for up-regulation of ODC is not at the transcriptional level, but may involve post-transcriptional modification(s) in lpr strains. The link between aberrant regulation of ODC and the immunopathogenesis of murine lupus nephritis indicates novel targets for lupus therapy.
Hsu
H C
HC
Program in Clinical Pharmacology, University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, New Brunswick 08903, USA.
Seibold
J R
JR
Thomas
T J
TJ
D012437:Adenosylmethionine Decarboxylase / Q000502:physiology; D000818:Animals; D001327:Autoimmune Diseases / Q000201:enzymology* / Q000235:genetics; D001483:Base Sequence; D015153:Blotting, Western; D005260:Female; D005786:Gene Expression Regulation / Q000276:immunology*; D007668:Kidney / Q000201:enzymology*; D007700:Kinetics; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D008969:Molecular Sequence Data; D009955:Ornithine Decarboxylase / Q000235:genetics*; D011073:Polyamines / Q000032:analysis; D014158:Transcription, Genetic
D011073:Polyamines; D009955:Ornithine Decarboxylase; D012437:Adenosylmethionine Decarboxylase