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AB0133 Urinary neutrophil gelatinase-associated lipocalin and monocyte chemoattractant protein-1 in a murine model of systemic lupus erythematosus
  1. S. Vagnani1,
  2. C. Tani1,
  3. L. Carli1,
  4. F. Querci1,
  5. R. Talarico1,
  6. C. Baldini1,
  7. A. Della Rossa1,
  8. S. Bombardieri1,
  9. M. Mosca1
  1. 1Rheumatology Unit, Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy


Background Neutrophil Gelatinase-Associated Lipocalin (NGAL) and Monocyte Chemoattractant Protein-1 (MCP-1) are promising biomarkers in lupus nephritis (LN). Recent data in human and mice systemic lupus erythematosus (SLE) highlight their potential involvement in LN pathogenesis; however controversial data exist on their role as marker of renal disease activity or damage.

Objectives The present study is aimed at evaluating urinary levels of NGAL and MCP-1 in a murine SLE model.

Methods Eight-weeks old female New Zealand Black/ White F1 mice (NZ) were used as SLE models and were compared to eight-weeks old C57BL6/J mice. In order to follow the disease course, body weight was periodically monitored and urinary protein excretion was weekly detected by dipstick analysis (Combur Test M; Roche). Weekly the urine samples were used to evaluate NGAL and MCP-1 levels by immunoassorbent assays (Mouse MCP-1 Elisa kit, Thermo Scientific; Mouse NGAL Elisa kit, Bioporto Diagnostics). At different disease stages (8, 18, 24, 30, 36, 40 weeks) the animals weresacrificed and peripheral blood serum samples were collected. Anti-dsDNA autoantibodies were analyzed by the enzyme immunoassorbent assay (ELISA) method using a commercial kit according to the manufacturer’s instructions (Mouse anti-dsDNA IgG ELISA Kit, Alpha Diagnostics International).

Results In the NZB/NZW F1 mice the average weight was 14.4 g at 8 weeks of age and it increased until week 36. The mean increase from week 8 to week 36 was 7.4 g. The mean urinary excretion of proteins was 72±34 mg/dl at 8 weeks of age and it increased in the course of the disease: 293±340 mg/dl and 624±469 mg/dl at 18 and 24 weeks of age. From week 8 to week 36 NGAL and MCP-1 urinary levels were not statistically significant while an important peak was detected on day 40: 2.00±0.94 ng/mland 5954±3921pg/ml respectively. In the NZB/NZW F1 mice serum titers of IgG auto-antibodies with specificity for dsDNA were negative until the 18th week (100% of the samples negative) becoming positive from the 24th week of age (70% of the samples positive). During the disease course no clinical and serological changes were observed in C57BL/6J mice; in particular, anti-dsDNA autoantibodies titers were negative in all experimental groups and urinary MCP-1 and NGAL concentrations did not increase in the observing time.

Conclusions The NZB/NZW F1 mice show abnormal clinical and serological parameters that tend to accrue over time during the disease course. While the appearance of anti-dsDNA autoantibodies is associated with increase in proteinuria, in our model urinary NGAL and MCP-1 seem to come after the first signs of the renal disease and they could represent late renal disease markers.

Disclosure of Interest None Declared

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