Mixed-Lipid Storage Disorder Induced in Macrophages and Fibroblasts by Oritavancin (LY333328), a New Glycopeptide Antibiotic with Exceptional Cellular Accumulation

ABSTRACT Oritavancin, a semisynthetic derivative of vancomycin endowed with a cationic amphiphilic character, accumulates to large extent in the lysosomes of eukaryotic cells (F. Van Bambeke, S. Carryn, C. Seral, H. Chanteux, D. Tyteca, M. P. Mingeot-Leclercq, and P. M. Tulkens, Antimicrob. Agents Chemother. 48:2853-2860, 2004). In the present study, we examined whether this accumulation could cause cell alterations in phagocytic (J774 mouse macrophages) and nonphagocytic (rat embryo fibroblasts) cells exposed to clinically meaningful (0- to 40-mg/liter) concentrations of oritavancin. Optical and electronic microscopy evidenced conspicuous alterations of the vacuolar apparatus in both cell types, characterized by the deposition of concentric lamellar structures, finely granular material, or other less-defined osmiophilic material, often deposed in giant vesicles. Biochemical studies showed an accumulation of phospholipids (1.5× control values) and free and esterified cholesterol (3 to 4× control values for total cholesterol). Accumulation of these lipids was in close relation to that of oritavancin (excess phospholipid/oritavancin and excess cholesterol/oritavancin molar ratios of 2 to 3 and 3 to 5, respectively). Cholesterol accumulation was rapid and reversible, and that of phospholipids was slower and poorly reversible. Vancomycin and teicoplanin, used as controls (50 and 100 mg/liter, respectively), did not cause any significant change in the lipid content of fibroblasts. The data therefore suggest that oritavancin has the potential to cause a mixed-lipid storage disorder in eukaryotic cells.