Nephrotoxic serum

Nephrotoxic serum should be kept frozen until used and then on ice until injected. 

Nephrotoxic serum (NTS) is prepared by immunizing male sheep with a lysate of rat glomeruli, isolated by differential sieving. This procedure has been described previously in great detail (5). 

Fig. 1. Scheme for induction of murine crescentic nephritis. (NSS, normal sheep serum NTS, nephrotoxic serum sc, subcutaneous iv, intravenous.)... 

Nephrotoxic serum should be titrated for each new preparation to assess disease severity. For each new batch inject groups of mice with NTS over a range of concentrations or volumes, and collect urine and kidney tissue on day 7. Compare degree of proteinuria and crescent score to ensure continuity between batches of NTS. We have chosen a dose of 50 pL of our current NTS, which induces between 50-60% crescent formation by day 7 in CD1 mice. An in-depth description of the production of NTS is provided in ref 5. 

Various experimental procedures were described as models for glomerulonephritis in human beings. Most of them were developed in rats and rabbits. They involve the reactions of antibodies against renal components, such as Masugi nephritis (Masugi and Sato 1934), Heymann nephritis (Heymann 1959), nephrotoxic serum nephritis (Unanue and Dixon 1967), crescentic type anti-glomerular membrane nephritis (Nagoe et al. 1994, 1998), anti-Thyl nephritis (Chen etal. 1999). 

Nephrotoxic serum nephritis is produced in animals by administration of heterologous antibody against... 

Male Wistar-Kyoto rats weighing 150 g receive either continuous administration of the test drug by an osmotic pump (ALZA Co., Palo Alto, USA) or saline. Twenty-four hours later, the rats are injected with 1 ml of nephrotoxic serum. At 9, 12, and 14 days, urine samples are collected and urinary protein levels are measured using the Lowry method. At 14 days the rats are sacrificed under ether anesthesia, and both kidneys are removed. Portions of these tissues are processed for light microscopy, immunofluorescence staining and immunoperoxidase staining. 

Kawasaki et al. (1992) induced crescentic glomerulonephritis with a small dose of nephrotoxic serum in WKY rats, which was characterized by the early infiltration of CD8 positive cells in glomeruli. In vivo depletion of CD8 positive cells completely prevented proteinuria and crescent formation. 

Suzuki S, Gejyo F, Kuroda T et al. (1998) Effect of a new elas-tase inhibitor, ONO-5046, on nephrotoxic serum nephritis in rats. Kidney Intern 53 1291-1208... 

Scharschmidt, L., Miller, M., Holthofer, H., Sinha, A., Schlondorff, D., Gibbons, N., and Santiago, A. (1990) A Fish Oil Diet Preserves Renal Function in Nephrotoxic Serum Nephritis, J. Lab. Clin. Med. 115,405M-14. 

Al-Refaie FN, Wickens DG, Wonke B, Kontoghiorghes GJ, Hoffbrand AV (1992) Serum non-transferrin-bound iron in beta-thalassaemia major patients treated with desferrioxamine and LI. Br J Haematol 82 431-436 Alfrey AC, Froment DH, Hammond WS (1989) Role of iron in the tubulo-interstitial injury in nephrotoxic serum nephritis. Kidney Int 36 753-759... 

The urinary excretion of jS-D-galactosidase during nephrotoxic serum glomerulonephritis and mercuric chloride-induced tubulonephritis in rats has been compared with that of lactate dehydrogenase, alkaline and acid phosphatases, and aspartate aminotransferase. ... 

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