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Use of a renal tubule cell line (LLC-PK1) to study the nephrotoxic potential of a kappa-type bence-jones protein
Authors:M-C. Diemert  V. Tricottet  L. Benel  G. Descamps  E. Escolano  J. Galli  M. Reynès  F. Rousselet
Affiliation:1. Laboratoire d’Immunochimie, Service du Dr Galli, Hopital de la Salpêtrière, 47, Boulevard de l’Hopital, 75651, PARIS Cedex 13, FRANCE
2. Laboratoire de Microscopie électronique, Université René Descartes-Paris V, 75006, Paris, France
3. Laboratoire de Biochimie métabolique et clinique Faculté des Sciences pharmaceutiques et biologiques, Université René Descartes-Paris V, 75006, Paris, France
4. GERC. Service de Biostatistiques et Informatique médicale H?pital Pitié-Salpêtrière, 47 Boulevard de l’H?pital, 75013, Paris, France
5. Service d’Anatomie et de Cytologie pathologiques H?pital Paul Brousse, 94804, Villejuif cedex, France
Abstract:Summary The cytotoxicity of a Bence-Jones protein was assessed using a porcine renal tubule cell line (LLC-PK1), with the aim of developing a model for studying the potential nephrotoxicity of these proteins. The effects of a kappa Bence-Jones protein on cell viability were studied by means of biochemical methods (supravital dye uptake and measurement of cellular enzyme activities) and morphological electron microscopy. After a 24-h-treatment with Bence-Jones protein, a moderate cytotoxicity (about 15%) was noted but only a minor difference compared to treatment with bovine albumin in the same conditions. The morphological study showed a few cells in the process of lysis, but their numbers were insufficient for the demonstration of a clear cytotoxic effect. Immunocytochemical studies showed Bence-Jones protein fixation on some cells, especially on the outer membrane. Labeling of the hyaloplasm and basal pole of a few cells pointed to internalization of protein by LLC-PK1 cells. Although the cytotoxicity of the Bence-Jones protein tested here was only moderate, the use of this model enabled its cytotoxic effect to be distinguished from that ofβ-lactoglobulin. This isolate could serve as a “moderate control” for a later study with a BJP having caused acute renal failure.
Keywords:Bence-Jones protein  LLC-PK1 cell line  cytotoxicity
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